HCV: Diagnosis and Natural History


1424

Early appearance of deep neutralizing antibody responses in persons clearing acute Hepatitis C infection revealed by a representative genotype 1 HCV E1E2 pseudoparticle library

William O. Osburn, Anna E. Snider, Brittany Wells, Rachel Latanich, Justin R. Bailey, David L. Thomas, Andrea Cox, Stuart Ray;

Medicine, Johns Hopkins University School of Medicine, Baltimore, MD

One of the obstacles to investigating the role of neutralizing antibodies (nAbs) in the outcome of acute HCV infection is the unique global and intrapersonal sequence variability of HCV, which complicates selection of a representative sequence. We used a representative genotype 1 HCV pseudoparticle (HCVpp) library to assess nAb responses during acute HCV infection to determine whether infection outcome is associated with the neutralizing antibody (nAb) timing or depth (neutralization of multiple genotype-matched variants). The HCVpp library comprises 19 genetically-distinct genotype 1 HCVpp derived from primary HCV isolates that represent the natural variability of genotype 1 E1E2 sequences. Neutralization of individual library HCVpp by the last viremic plasma sample obtained before clearance was compared to infection duration-matched specimens obtained from subjects who developed persistent infection. Overall, 60.3% of plasma samples neutralized at least one HCVpp in the genotype 1 HCVpp library while the subtype 1a and 1b HCVpp with the highest neutralization sensitivity was neutralized by only 41.3% and 39.7% of subjects, respectively, (subtype 1a, P = 0.05, subtype 1b, P = 0.03) highlighting the utility of using a library screening approach for assessing nAb responses. A trend toward an increased percentage of Clearance subjects neutralizing at least one HCVpp compared to Persistence subjects was observed (76.2% versus 52.4%, P = 0.101). However, the median number of library HCVpp neutralized by Clearance subjects was six-fold greater than Persistence subjects (6 versus 1, P = 0.007). Moreover, the number of library HCVpp neutralized by Persistence subjects increased during acute infection (P < 0.001) while the number of library HCVpp neutralized by Clearance subjects decreased following control of viremia (P = 0.03). Surprisingly, the depth of anti-genotype 1 nAb responses was not different in subjects infected with genotype 1, 2 or 3 viruses (P = 0.78). Interestingly, two single nucleotide polymorphisms in the HLA-DQ locus were associated with nAb depth. Taken together, these data demonstrate that nAb responses are delayed in persistently infected individuals then progressively deepen, whereas in persons who control viremia deep responses are detected early and contract after clearance of viremia, independent of the infection viral genotype. These findings provide further evidence for the role of nAb in controlling HCV infection and highlight the potential benefit of generating deep anti-HCV nAb responses by vaccination.

Disclosures:

David L. Thomas - Grant/Research Support: Merck, Gilead

Stuart Ray - Advisory Committees or Review Panels: Abbott Laboratories, Boer-hinger Ingelheim

The following people have nothing to disclose: William O. Osburn, Anna E. Snider, Brittany Wells, Rachel Latanich, Justin R. Bailey, Andrea Cox

1425

Comparison of the overall survival between patients with HCV-induced advanced hepatic fibrosis and the general population

Adriaan J. van der Meer1, Bettina E. Hansen1, Jordan J. Feld2, Heiner Wedemeyer3, Jean-Francois Dufour4, Frank Lammert5, Andres Duarte-Rojo2, Michael P. Manns3, Stefan Zeuzem6, Wolf P. Hofmann6, Robert J. de Knegt1, Bart J. Veldt1, Harry L. Janssen1,2;

1Gastroenterology & Hepatology, Erasmus MC, Rotterdam, Netherlands; 2Liver Centre, Toronto Western Hospital, Toronto, ON, Canada; 3Department of Gastroenterology, Hepatology, and Endocrinology, Medical School Hannover, Hannover, Germany; 4Department of Gastroenterology, Hepatology, and Endocrinology, University of Bern, Bern, Switzerland; 5Department of Medicine II, Saarland University Medical Center, Homburg, Germany; 6Medizinische Klinik 1, Klinikum der Johann Wolfgang Goethe-Universität, Frankfurt am Main, Germany

INTRODUCTION Sustained virological response (SVR) is the primary efficacy measure for the treatment of chronic hepatitis C virus (HCV) infection, but randomized controlled trials showing a clinical benefit of antiviral therapy and validating SVR as surrogate endpoint are lacking. We compared the overall survival of patients with HCV-induced advanced fibrosis, with and without SVR, to that of the general population. METHODS Survival was assessed in an international cohort of consecutive patients with chronic HCV infection and advanced fibrosis (Ishak score 4-6) who started interferon-based therapy between 1990 and 2003. Per virological response group, the observed survival among patients was compared to the expected survival from matched age-, gender- and calendar time-specific death rates of the general Dutch population using the life table method and Wilcoxon (Gehan) test. RESULTS In total, 530 patients were followed for a median of 8.4 (IQR 6.4-11.4) years. Median age at baseline was 48 (IQR 42-56) years and 369 (70%) patients were male. SVR was attained by 192 (36%) patients. Cox regression analysis showed SVR (included as time-dependent variable) was independently associated with reduced mortality (adjusted HR 0.24, 95%CI 0.14-0.49, p<0.001). The cumulative 10-year survival was 74.0% (95%CI 71.6-79.8) among patients without SVR, which was significantly lower compared to the age- and gender-matched general population (p<0.001). Patients with SVR showed a cumulative 10-year survival of 91.1% (95%CI 85.5-96.7), which did not differ significantly from the standardized general population (p=0.571). CONCLUSION Despite established advanced fibrosis or cirrhosis, patients with chronic HCV infection who attained SVR show a comparable survival to that of a general population. This further supports SVR is a relevant surrogate endpoint of anti-HCV therapy.

image

Disclosures:

Adriaan J. van der Meer - Speaking and Teaching: MSD

Jordan J. Feld - Advisory Committees or Review Panels: Roche, Merck, Vertex, Gilead, Abbott, Tibotec, Theravance, Achillion; Speaking and Teaching: Merck, Roche, Abbott

Heiner Wedemeyer - Advisory Committees or Review Panels: Transgene, MSD, Roche, Gilead, Abbott, BMS, Falk; Grant/Research Support: MSD, Novartis, Gilead, Roche, Abbott; Speaking and Teaching: BMS, MSD, Novartis, ITF

Jean-Francois Dufour - Advisory Committees or Review Panels: Bayer, BMS, Gilead, Jansse, Novartis, Roche

Michael P. Manns - Consulting: Roche, BMS, Gilead, Boehringer Ingelheim, Novartis, Idenix, Achillion, GSK, Merck/MSD, Janssen, Medgenics; Grant/Research Support: Merck/MSD, Roche, Gilead, Novartis, Boehringer Ingelheim, BMS; Speaking and Teaching: Merck/MSD, Roche, BMS, Gilead, Janssen, GSK, Novartis

Stefan Zeuzem - Consulting: Abbvie, Achillion Pharmaceuticals, Boehringer Ingelheim GmbH, Bristol-Myers Squibb Co., Gilead, Novartis Pharmaceuticals, Merck & Co., Idenix, Janssen, Roche Pharma AG, Vertex Pharmaceuticals, Presidio, Santaris, Inc

Robert J. de Knegt - Advisory Committees or Review Panels: MSD, Roche, Norgine, Janssen Cilag; Grant/Research Support: Gilead, MSD, Roche, Janssen Cilag, BMS; Speaking and Teaching: Gilead, MSD, Roche, Janssen Cilag

Bart J. Veldt - Board Membership: GSK

Harry L. Janssen - Consulting: Abbott, Bristol Myers Squibb, Debio, Gilead Sciences, Merck, Medtronic, Novartis, Roche, Santaris; Grant/Research Support: Anadys, Bristol Myers Squibb, Gilead Sciences, Innogenetics, Kirin, Merck, Medtronic, Novartis, Roche, Santaris

The following people have nothing to disclose: Bettina E. Hansen, Frank Lammert, Andres Duarte-Rojo, Wolf P. Hofmann

1426

Polymorphisms consisting of (TA)n dinucleotide repeat near IL28B gene could improve the predictive value for HCV spontaneous clearance with IL28B SNPs

Masaya Sugiyama1, Satoshi Hiramine2, Norihiro Furusyo2, Akio Ido3, Hirohito Tsubouchi3, Hisayoshi Watanabe4, Yoshiyuki Ueno4, Masaaki Korenaga1, Kazumoto Murata1, Naohiko Masaki1, Masashi Mizokami1;

1National Center for Global Health and Medicine, Ichikawa, Japan; 2Kyushu University, Fukuoka, Japan; 3Kagoshima University, Kagoshima, Japan; 4Yamagata University, Yamagata, Japan

Background; Recent genome-wide-association studies revealed that single nucleotide polymorphisms (SNPs) around interleukin (IL)-28B were associated with response to interferon (IFN) therapy of the anti-hepatitis C virus (HCV) therapy and with spontaneous HCV clearance. We have also reported that the strong association between IL28B SNPs and IFN therapy in Japanese. The favorable genotype of IL28B has been also associated with spontaneous clearance. However, the effect of IL28B SNPs was lower than the results observed in the study of IFN therapy, suggesting that other factors might be involved in HCV spontaneous clearance. We previously reported that the length of the TA dinucleotide repeat in the promoter region of IL28B has a wide variation, and the transcriptional activity increased gradually in a TA repeat length-dependent manner. In the present study, we determined the length of the TA repeat to investigate the correlation to with HCV spontaneous clearance. Methods; Total 2041 Japanese genomic samples were enrolled (274 with HCV spontaneous clearance, 457 with chronic HCV infection and 1310 healthy donors). IL28B SNPs were genotyped using Invader assay. The length of TA repeat was determined by 31 30xl sequencer and GeneMapper software. The association between clinical and genetical data was statistically analyzed using STATA software. Results; The variation of TA repeat number ranged from 1 0 to 18, and the most frequent repeat was 12 (83.4%) in the population. A univariate analysis showed significant differences between groups with HCV spontaneous clearance and chronic infection in sex (men 51.4% vs. women 67.0%), age (68.1 ± 11.1 vs. 64.0 ± 10.8 years), IL28B (rs8099917) genotypes (TT 92.0% vs. non-TT 67.4%), and TA repeat number (≧12/12, 99.6% vs. <12/11, 95.0%) (all P<0.01). Multiple logistic regression model showed women (OR, 2.05; 95%CI, 1.47-2.87), older age (OR, 1.03; 95%CI, 1.01-1.04), IL28B (rs8099917) genotypes TT (OR, 5.40; 95%CI, 3.31-8.80), and TA repeat number ≧12/12 (OR, 10.7; 95%CI, 1.40-82.4) as independently significant factors for HCV spontaneous clearance. Analyzing these 4 factors by decision tree model, among women aged ≧68 years with IL28B (rs8099917) TT, those with TA repeat genotype ≧12/12 had a high probability of spontaneous clearance. Conclusions; TA repeat number ≧12/12 in the promoter region of IL28B was associated to HCV spontaneous clearance. It could be the novel genetic factor to improve the predictive value for HCV clearance with IL28B SNPs.

Disclosures:

Hirohito Tsubouchi - Grant/Research Support: MSD, Chugai Pharmaceutical, Kan Research Institute, Daiichi-Sankyo, Eisai, Tanabe Mitsubishi; Speaking and Teaching: MSD, Tanabe Mitsubishi

Yoshiyuki Ueno - Advisory Committees or Review Panels: Jansen

The following people have nothing to disclose: Masaya Sugiyama, Satoshi Hiramine, Norihiro Furusyo, Akio Ido, Hisayoshi Watanabe, Masaaki Korenaga, Kazumoto Murata, Naohiko Masaki, Masashi Mizokami

1427

Predicting Utilization of Liver Transplantation for Hepatitis C-related Liver Disease in the United States

Archita P. Desai1, Gary L. Davis3, Nancy Reau2, Donald M. Jensen2;

1Department of Medicine, Section of Gastroenterology, Hepatology and Nutrition, University of Chicago Medical Center, Chicago, IL; 2Department of Medicine, Center for Liver Diseases, University of Chicago Medicine, Chicago, IL; 3Department of Internal Medicine, Division of Hepatology, Baylor University Medical Center, Dallas, TX

Background: By all estimates, the burden of morbidity and mortality due to hepatitis C (HCV) will increase over the next two decades in the US as more HCV-infected persons develop cirrhosis and its complications. As anti-viral therapies (AVT) become increasingly successful and accessible, their impact on the utilization of liver transplantation (LT) is likely to change. Furthermore, the effect of birth-cohort screening (BCS) on LT utilization is unclear. While increasing prevalence of HCV-cirrhosis may increase demand for LT, we hypothesize this need will be partly offset by the increasing success of AVT. Aim: We report forecasts of future LT utilization that consider the combined effects of identification of new cases through BCS and intervention with more effective AVT. Methods: We used a previously developed multicohort natural history model to simulate progression of patients predicted to have advanced fibrosis and cirrhosis starting in the year 2015 and ending in 2025. We adjusted previous estimates of cirrhosis prevalence based on success of BCS (50% vs. 100% undiagnosed cases identified). Medical literature informed our best estimates of moving between disease stages with and without sustained virologic response (SVR). We then modified the model to estimate the impact of varying treatment uptake rates (25%, 50%, 75%, 100%). Finally, we used SVR rates in cirrhotic and post-transplant patients consistent with anticipated interferon-free regimens(80% to 90%). Results: Assuming that half of the undiagnosed HCV patients could be identified by BCS, 1 million cirrhotic patients would be eligible for treatment and disease management in 2015. In sensitivity analysis, the success of BCS, AVT efficacy, and treatment uptake rates all significantly impact disease outcome and need for LT. Based on initial analysis, we estimate a 10% decline in need for LT if BCS is able to identify 100% of cases of cirrhosis compared to 50% identification. Furthermore, compared to current standard of care, if interferon-free therapy is applied to 50% vs. 100% of treatment-eligible cirrhotics, need for LT would decline by 20% vs. 55%. These factors plus the potential of competing risk due to comorbidities amongst the aging HCV population all predict a decreased need for donors for HCV patients over the next 1 0 years. Conclusions: Given predicted SVR rates of 80%-90% in patients with advanced fibrosis, prior predictions of LT utilization are no longer accurate. Understanding the implications of improved AVT combined with BCS in this population will inform campaigns to improve both screening and treatment uptake in a traditionally under-served population.

Disclosures:

Gary L. Davis - Consulting: Genentech; Grant/Research Support: Vertex, Genen-tech, Tibotec, Abbott, Boehringer Ingelheim, Bristol Myers Squibb, Gilead, Novaris, Pharmasett

Nancy Reau - Advisory Committees or Review Panels: Genentech, Kadmon, Jannsen, Vertex, Idenix; Consulting: IHEP; Grant/Research Support: Vertex, Gilead, abbott

Donald M. Jensen - Advisory Committees or Review Panels: Abbvie, Boehringer, BMS, Genentech/Roche, Merck, Gilead, Janssen; Grant/Research Support: Abb-vie, Boehringer, BMS, Genentech, Janssen, Gilead

The following people have nothing to disclose: Archita P. Desai

1428

Semi-automated digital scan technology allows fibrosis measurement in small liver biopsy samples that accurately correlates with clinical outcomes

Yi Huang1,2, Bastiaan de Boer3, Leon Adams1,2, Gerry C. Mac-Quillan1,2, Max K. Bulsara4, Gary P. Jeffrey1,2;

1School of Medicine and Pharmacology, University of Western Australia, Perth, WA, Australia; 2Department of Gastroenterology and Hepatology, Sir Charles Gairdner Hospital, Perth, WA, Australia; 3Department of Anatomical Pathology, PathWest, QEII Medical Centre, Perth, WA, Australia; 4Institute of Health and Rehabilitation Research, University of Notre Dame, Perth, WA, Australia

Background: Severity of liver fibrosis correlates with adverse clinical outcomes. Histopathological scoring systems mainly assess architectural abnormalities and need a minimum biopsy size (≥10mm). Quantification of liver collagen has the potential to use small size biopsies and improve the prediction of clinical outcomes. Aim: To test the ability of collagen proportional area (CPA) to predict clinical outcomes for chronic hepatitis C (CHC) patients and compare it with Metavir stage. Methods: Clinical outcomes were determined using population based data-linkage methodology for chronic hepatitis C (CHC) patients from 1992-2012. Quantitative digital image analysis was used to measure CPA. Results: 533 patients with CPA measurement area >5 mm2 were included. Median follow up was 10.5 years and 26 developed HCC, 39 developed liver decompensation and 33 had a liver related death (LRD). 1 02 had Metavir F0, 244 had F1, 89 had F2, 48 had F3 and 50 had F4. CPA values ranged from 1.3%-44.6%. CPA was correlated with Metavir stage (r=0.615, P<0.001). Univariate analysis found CPA, Metavir stage and age were significantly associated with decompensation, HCC and LRD. Multivariate analysis found CPA and Metavir stage were independently associated with decompensation and LRD while Metavir stage and age were significantly associated with HCC. CPA stage (C1: 0%-5%, C2: 5%-10%, C3: 10%-20%, C4: >20%) was used to stratify risk. There was a significant difference in composite end point free survival (HCC, decompensation and LRD) between C1 and C2 (p=0.010), C2 and C3 (p<0.001), C3 and C4 (p<0.001). The 15 year composite end point free survival probability was 97.1% for C1, 88.7% for C2, 60.5% for C3, 7.3% for C4. A significant difference was also found in separate analyses for HCC development between C1 and C2 (p=0.016), C2 and C3 (p<0.001), C3 and C4 (p=0.004) and for decompensation between C2 and C3 (p=0.01 0), C3 and C4 (p<0.001) and for LRD between C2 and C3 (p=0.0002) and C3 and C4 (p<0.001). The only significant difference between Metavir stages was between F3 and F4 for the composite end point and all three endpoints (p<0.001). Among cirrhotic patients C4 had significantly worse LRD than C1-C3 (p=0.026). For non-cirrhotic patients C1 had significantly better HCC free survival than C2-C4 (p=0.006). Cox regression found no significant interaction between biopsy size and CPA predictive ability. Conclusions: Simple digital technologies allowed measurement of CPA in previously inadequate sized liver biopsy samples. CPA stage was superior to Metavir stage in its ability to stratify risk of LRD, HCC and liver decompensation for CHC patients.

Disclosures:

Gary P. Jeffrey - Advisory Committees or Review Panels: MSD, Novartis

The following people have nothing to disclose: Yi Huang, Bastiaan de Boer, Leon Adams, Gerry C. MacQuillan, Max K. Bulsara

1429

Impact of patatin-like phospholipase domain-containing protein 3 (PNPLA3) polymorphism on steatosis and fibrosis in patients with chronic hepatitis C treated with pegylated interferon plus ribavirin

Naoto Kawabe, Senju Hashimoto, Masao Harata, Yoshifumi Nitta, Michihito Murao, Takuji Nakano, Hiroaki Shimazaki, Yuko Arima, Toshiki Kan, Nakaoka Kazunori, Masashi Ohki, Takagawa Yuka, Toru Nishikawa, Keisuke Osakabe, Naohiro Ichino, Kentaro Yoshioka;

Department of Liver, Biliary Tract and Pancreas Diseases, Fujita Health University, Toyoake, Japan

Background/Aim: The single nucleotide polymorphism (SNP) rs738409 (C>G) in patatin-like phospholipase domain-containing protein 3 (PNPLA3) has been reported as a strong genetic determinant of hepatic fat accumulation, and to influence fibrosis severity in patients with non-alcoholic fatty liver disease. The aim of this study was to investigate the impact of PNPLA3 polymorphism on hepatic steatosis, fibrosis and therapeutic effects in patients with chronic hepatitis C (CHC) treated with pegylated interferon (PEG-IFN) plus ribavirin (RBV). Methods: We performed a retrospective cohort study involving consecutive 133 CHC patients (58 males, 75 females; mean age 55.7) infected with hepatitis C virus (HCV) genotype 1, treated with PEG-IFN plus RBV. The PNPLA3 rs738409 SNP and HCV core mutation (aa70, 91) were genotyped. The inflammation and fibrosis were evaluated in the liver biopsy specimen with inflammation and fibrosis score assessed by the METAVIR criteria. The proportion of the area of steatosis was measured quantitatively and objectively by image analysis software (Image-Pro Plus) in the liver biopsy specimen. Results: The proportion of the area of steatosis was significantly higher in patients with HCV core aa70 mutant (0.626%) than those with aa70 wild-type (0.202%) (P=0.035). The proportion of the area of steatosis was also significantly higher in patients with GG/CG genotype (N=92, 0.322%) of PNPLA3 rs738409 SNP than those with CC genotype (N=41, 0.189%) (P=0.013). The inflammation grade was significantly higher in patients with GG/CG genotype (A0-A1/A2-A3: 36/56) of PNPLA3 rs738409 SNP than those with CC genotype (A0-A1/A2-A3: 24/17) (P=0.038). The fibrosis stage was significantly higher in patients with GG/CG genotype (F0-F2/F3-F4: 43/49) of PNPLA3 rs738409 SNP than those with CC genotype (F0-F2/F3-F4: 27/14) (P=0.042). PNPLA3 rs738409 SNP was not associated with the sustained virological response (SVR) rate of PEG-IFN plus RBV treatment (CC; 30%, CG/GG; 39%). Conclusion: The PNPLA3 rs738409 SNP and HCV core mutation are associated with steatosis in CHC patients. The PNPLA3 rs738409 SNP is also associated with inflammation and fibrosis in CHC patients, but does not influence on the outcome of PEG-IFN plus RBV treatment.

Disclosures:

Kentaro Yoshioka - Grant/Research Support: Chugai, Schering-Plough, Bristol Myers Squibb, Tanabe Mitsubishi, Taiho, Otsuka, Ajinomoto, Tore Medical, Torii, Boston,,ÄÄScientific

The following people have nothing to disclose: Naoto Kawabe, Senju Hashimoto, Masao Harata, Yoshifumi Nitta, Michihito Murao, Takuji Nakano, Hiroaki Shimazaki, Yuko Arima, Toshiki Kan, Nakaoka Kazunori, Masashi Ohki, Takagawa Yuka, Toru Nishikawa, Keisuke Osakabe, Naohiro Ichino

1430

A Polymorphism in the EGF Gene Independently Predicts Hepatic Outcomes in HCV-Related Cirrhosis

Lindsay Y. King1, Kara B. Johnson2,1, Hui Zheng3, Lan Wei4, Thomas Gudewicz5, Ajayi Tokunbo6, Nneka Ufere1, Kenneth Tan-abe4, Bryan C. Fuchs4, Raymond T. Chung1;

1Medicine, Division of Gastroenterology, Massachusetts General Hospital, Boston, MA; 2Harvard Medical School, Boston, MA; 3Biostatistics Center, Massachusetts General Hospital, Boston, MA; 4Surgery, Massachusetts General Hospital, Boston, MA; 5Pathology, Massachusetts General Hospital, Boston, MA; 6Medicine, Salem Hospital, Salem, MA

Background and Aims: Several SNPs including rs4444903 in the epidermal growth factor (EGF) gene, rs738409 in the patatin-like phospholipase-3 (PNPLA3) gene and rs1 2979860 near the IL28B gene have been variably associated with treatment response, fibrosis, steatosis, and development of hepatocellular carcinoma (HCC) in chronic hepatitis C. No study has comprehensively examined the effects of these SNPs on natural history of HCV cirrhosis. We tested the association between clinical, demographic, and genetic data and outcomes in patients with HCV cirrhosis. Methods: We performed a retrospective cohort study of 169 subjects with HCV and biopsyproven cirrhosis who did not have evidence of decompensation or HCC at the time of biopsy. Genotyping was performed on formalin fixed paraffin embedded liver tissue specimens. Cox proportional hazards model was used to determine the hazard ratio for risk of decompensation (ascites, encephalopathy, variceal bleed, HCC, liver death). Results: During a median follow up of 6.6 years, 66 patients experienced a decompensation event. EGF AA, PNPLA3 CC and IL28B CC were each associated with a decreased decompensation risk in univariate analysis. In multivariable Cox regression modeling, EGF AA genotype was associated with a significantly decreased risk of decompensation (HR = 0.34, p=0.03) even after adjusting for other univariate significant predictors (Table). Conclusions: HCV cirrhotics with the EGF AA genotype, a functional polymorphism associated with decreased EGF production, have a decreased risk of clinical outcomes. These data imply that EGF genotyping will have utility in identifying persons at risk for disease progression. Further study of the predictive value of EGF genotyping in patients with earlier stages and other etiologies of liver disease is warranted.

Table 1. 
 Unadjusted HR (95% CI)p valueMultivariable HR (95%CI)P value
  1. Age, BMI, sex, race, alcohol use, tobacco use, diabetes, viral load, ALT, and total bilirubin were not significant in univariate analysis.

EGF AA0.30(0.15-0.61)0.00090.34(0.13-0.88)0.03
Genotype (1,4,6 versus 2,3)2.62(1.21-5.65)0.013.72(1.25-11.08)0.02
Creatinine mg/dL1.72(0.98-2.99)0.062.14(1.26-3.64)0.005
Platelet (/I0,000/mm3 increase)0.86(0.82-0.91)<0.00010.90 (0.83-0.96)0.003
AST (/10U/L increase)1.05(1.02-1.08)0.00071.06(1.02-1.11)0.002
Incomplete cirrhosis0.49(0.29-0.81)0.005  
PNPLA3 CC0.56 (0.34-0.90)0.02  
IL28B CC0.56 (0.33-0.95)0.03  
SVR0.13(0.04-0.41)0.0005  
Albumin g/dL0.47 (0.30-0.74)0.001  

Disclosures:

Kenneth Tanabe - Patent Held/Filed: Massachusetts General Hospital

Raymond T. Chung - Advisory Committees or Review Panels: Idenix; Consulting: Enanta; Grant/Research Support: Gilead, Merck, Mass Biologic, Gilead

The following people have nothing to disclose: Lindsay Y. King, Kara B. Johnson, Hui Zheng, Lan Wei, Thomas Gudewicz, Ajayi Tokunbo, Nneka Ufere, Bryan C. Fuchs

1431

Estimation of risk for the development of hepatocellular carcinoma after successful eradication of hepatitis C by anti-viral therapy

Masayuki Kurosaki, Kaoru Tsuchiya, Yutaka Yasui, Nobuharu Tamaki, Takanori Hosokawa, Namiki Izumi;

Musashino Red Cross Hospital, Tokyo, Japan

Background and Aims: Highly effective antiviral therapy will lead to increased number of patients with successful eradication of hepatitis C (HCV). While the overall incidence of hepatocellular carcinoma (HCC) is expected to decrease, the number of HCC development after HCV eradication may paradoxically increase. The aim of the present study was to estimate risk factors for development of HCC following successful eradication of HCV. Methods: Multicenter study was conducted by Japanese Red Cross Liver Study Group, involving 18 hospitals and medical centers nationwide. A total of 785 genotype 1b chronic hepatitis C patients who had successful eradication of HCV by interferon therapy from 1992 to 2009 were enrolled. Median period of follow up was 6.5 years. Data were collected before and at 24 weeks after the completion of interferon therapy. Results: The cumulative incidence of HCC development after HCV eradication was 4.0 % in 3-yr, 7.1 % in 5-yr, and 12.4% in 7-yr, respectively. Multivariable Cox regression analysis revealed that age over 60 (HR 3.80), male (HR 3.54), platelet counts below 150 (109/L) (HR 2.1 1), and serum alpha-fetoprotein (AFP) > 5ng/ml at 24 weeks after the completion of interferon therapy (HR 3.21) were independent risk factors for the development of HCC. The 5 year incidence of HCC in patients with AFP levels of <5, 5-9, 10-19, and ≥ 20 ng/ml was 3.7%, 12.2%, 21.3%, and 34.5% respectively. A proportion of patients with AFP levels of <5 ng/ml at 24 weeks after the completion of interferon therapy was 69%. Among them, 89% had persistently low AFP levels up to 3 years. The cumulative incidence of HCC development beyond 5 years was significantly lower in those patient with persistently low AFP levels up to 3 years compared to others: cumulative HCC incidence was 2.0% vs. 14.9% in 7-yr, and 2.0% vs. 28.4% in 10-yr, respectively (p<0.0001). The incidence of persistently low AFP levels were smaller in patients older than 55 (65% vs. 73%, p=0.03) and in patients with advanced fibrosis (METAVIR F3-4) (54% vs. 73%, p=0.0003). Duration of interferon therapy or the use of ribavirin was not associated with the incidence of persistent AFP normalization. Conclusions: Older patients with advanced fibrosis are less likely to have persistent AFP normalization after successful eradication of HCV. Older age, advanced fibrosis (as reflected by lower platelet counts) and high AFP levels after HCV eradication are hallmarks of residual risk for HCC development, and patients with these factors may be the candidate for a careful HCC surveillance even after the complete eradication of hepatitis C virus.

Disclosures:

Namiki Izumi - Speaking and Teaching: MSD Co., Chugai Co., Daiichi Sankyo Co., Bayer Co., Bristol Meyers Co.

The following people have nothing to disclose: Masayuki Kurosaki, Kaoru Tsuchiya, Yutaka Yasui, Nobuharu Tamaki, Takanori Hosokawa

1432

Association of IL-28B polymorphisms with the natural history of Hepatitis C in an Irish cohort

Carthage Moran1, Susan Corbett1, Elizabeth Kenny-Walsh1, Liam J. Fanning2, Orla M. Crosbie1;

1Hepatology, Cork University Hospital, Corlk, Ireland; 2Molecular Virology Laboratory, University College Cork, Cork, Ireland

Our study aimed to investigate the association of IL-28B polymorphisms with the natural history of Hepatitis C Virus (HCV) in a female population in Ireland, infected with HCV genotype 1 b via contaminated anti-D immunoglobulin. 120 HCV antibody positive patients, were identified retrospectively from the hospital HCV database; HCV PCR status and genotype was already measured by the Molecular Virology Labarotory at Cork University Hospital. IL-28B was measured on all patients following consent and standard venepuncture. Single Nucleotide Polymorphism (SNP) analysis was performed by KBioSciences, UK. Statistical analysis included Fisher's Exact test to calculate p-val-ues on categorical variables, odd's ratio and confidence intervals. All 120 patients were female, had the same genotype and method of contamination. 72 patients (60% of cohort) had persistent HCV infection. Table 1 shows significant differences in rs 12979860 allelle (C/C vs C/T, T/T and C/T +T/T) frequencies between the group that spontaneously cleared the virus, and those that remained PCR positive. Patients with the C/C genotype were fifteen times more likely to clear HCV relative to patients with the C/T and T/T genotypes combined (OR= 15.2, CI 6-37.8, p =0.0001). Analysis of rs 8099917 (T/T vs G/G, T/G, G/G + T/G) shows a significant difference of the above frequencies between the group that spontaneously cleared the virus, and those that remained PCR positive except for T/T vs G/G (recognising small sample size). Patients with T/T genotype were ten times more likely to clear HCV relative to patients with the T/G and G/G genotypes combined. Our study concurs with the global association between IL-28 genotypes and the clearance of HCV. The C/C genotype (rs 12979860) and T/T genotype (rs809991 7) may help in predicting patients who spontaneous clear HCV following receipt of contaminated anti-D immunoglobulin.

Table 1

Genotype% clearance% persistanceComparisonOdds Ratiop-value
rs12979860     
T/T16.6783.33C/C versus T/T12.2 (1.3-112)0.0156
C/T13.5686.44C/C versus C/T15.5(6-40)0.0001
T/T + C/T13.8586.15C/C versus C/T + T/T15.2(6-37.8)0.0001
C/C70.9129.09   
rs8099917     
G/G2575T/T versus G/G4.6 (0.4-46)0.3012
T/G12.587.5T/T versus T/G10.6(3.9-28)0.0001
G/G + T/G13.4686.54T/T versus G/G + T/G9.7 (3.8-24.8)0.0001
T/T60.2939.71   

Disclosures:

The following people have nothing to disclose: Carthage Moran, Susan Corbett, Elizabeth Kenny-Walsh, Liam J. Fanning, Orla M. Crosbie

1433

Probabilities of Survival, Liver Transplantation, and Liver Complications by Hepatic Fibrosis Stage among Patients with Chronic Hepatitis C Virus (HCV) Infection from Chronic Hepatitis Cohort Study (CHeCS)

Fujie Xu1, Jia Li2, Talan Zhang2, Loralee B. Rupp2, Mei Lu2, Anne C. Moorman1, Stuart C. Gordon2, Philip R. Spradling1, Eyasu H. Teshale1, Joseph A. Boscarino3, Vinutha Vijayadeva4, Mark A. Schmidt5, Scott D. Holmberg1;

1Division of Viral Hepatitis, CDC, Atlanta, GA; 2Henry Ford Hospital, Detroit, Detroit, MI; 3Geisinger Health System, Danville, PA; 4Kaiser Permanente Hawaii, Honolulu, HI; 5The Center for Health Research, Kaiser Permanente Northwest, Portland, OR

Background: HCV-infected patients with more advanced fibrosis are at risk of hepatic decompensation. Fibrosis is reversible and new antiviral therapies have increased treatment alternatives. Using data from CHeCS, an ongoing observational cohort study among patients receiving care at 4 integrated healthcare systems in the U. S., we sought to examine associations between fibrosis stage (via liver biopsy and biomarker score) and clinical outcomes. Methods: Patients with confirmed HCV mono-infection were classified into 3 fibrosis stages based on biopsy results ranked as F4, cirrhosis; F3, numerous septa without cirrhosis; or lower (F0-2). Using cutoff points mapped to biopsy results, patients were grouped based on FIB-4 score. Clinical endpoints of survival, liver transplantation, and diagnoses of hepatocellular carcinoma (HCC) or ascites were ascertained using ICD-9 codes and chart review. The 5-year probability of each clinical endpoint during 2006-2010 was estimated using extended Kaplan-Meier by fibrosis stage over time. Cox regression models were used to adjust for demographic and clinical covariates at baseline. Results: Of 2,384 patients (mean age 54 yrs, 61% male, 58% white) with biopsy results available, 5-year survival rates ranged from 81% to 97% by fibrosis stage (Table). In persons of F4 stage based on biopsy (mean age 59 yrs, treatment naïve 46%), 23% had liver transplantation, 17% had HCC and 18% developed ascites within 5 years. The 5-year survival probability was 57% in patients with FIB4>5.88, but 96% in those with FIB4<1.21. Of 5,569 patients (mean age 54 yrs, treatment naïve 83%) with baseline FIB4<1.21, none had liver transplantation or HCC/ascites within 5 years. Compared to those with FIB4<1.21, FIB4>5.88 was associated with higher risk of death (adjusted hazard ratio (aHR) 3.1), liver transplantation, HCC and ascites (all aHR >8.2). Conclusions: Fibrosis stage based on either biopsy or FIB4 was strongly associated with probabilities of survival and liver-related complications. Fibrosis stage F4 and FIB4>5.88 were associated with higher morbidity and mortality, whereas those with FIB4<1.21 appear to have excellent 5-year prognoses.

Table. 5-year probabilities (95% CI) of clinical endpoints by fibrosis stage.

EndpointsBiopsy stageFIB4 group
F4F3F0-2>5.881.21 to 5.88<1.21
(n=457)(n=299)(n=1628)(n=1907)(n=8324)(n=5569)
Survival0.81(.77-.85)0.90(.86-.94)0.97(.96-.98)0.57(.57-.60)0.90(.89-.90)0.96(.96-.97)
Liver transplant0.23(.19-.27)0.02(0-.04)0.01(0-.01)0.12(.ll-.14)0.02(.01-.02)0
HCC0.17(.13-.21)0.03(.01-.05)0.0K.01-.020.06(.05-.07)0.01(.01-.01)0
Ascites0.18(.l3-.22)0.06(.03-.09)0.02(.01-.02)0.17(.15-.19)0.02(.02-.02)0

Disclosures:

Stuart C. Gordon - Advisory Committees or Review Panels: Tibotec; Consulting: Merck, CVS Caremark, Gilead Sciences, BMS; Grant/Research Support: Roche/Genentech, Merck, Vertex Pharmaceuticals, Gilead Sciences, BMS, Abbott, Intercept Pharmaceuticals, Exalenz Sciences, Inc.

The following people have nothing to disclose: Fu ie Xu, Jia Li, Talan Zhang, Loralee B. Rupp, Mei Lu, Anne C. Moorman, Phi ip R. Spradling, Eyasu H. Teshale, Joseph A. Boscarino, Vinutha Vijayadeva, Mark A. Schmidt, Scott D. Holmberg

1434

Ten-year prognostic performances of FibroTest (FT) and transient elastography (TE) in patients with chronic hepatitis C (CHC)

Thierry Poynard1, Julien Vergniol2, Yen Ngo3, Juliette Foucher2, Mona Munteanu3, Wassil Merrouche2, Vincent Thibault1, Olivier Deckmyn3, Pascal Lebray1, Vlad Ratziu1, Victor de Ledinghen2;

1Groupe Hospitalier Pitié-Salpêtrière, Paris, France;2Bordeaux University, Bordeaux, France; 3BioPredictive, Paris, France

Background and aim 5-year performances of FT and TE have been validated in CHC in 2 prospective cohorts, (Ngo 2006 and Vergniol 201 1) for survival [overall survival (OS), and survival without liver related complications (S-LRC)]. The long-term prognostic values on each LRC are unknown due to the limited sample size and follow-up. Patients and methods To increase the power, we pooled the updated individual data of these cohorts at 10 years. Patients (pts) with CHC were included if at least 1 FT and 1 TE were performed, and excluded if they had other cause of liver disease. The main endpoints (estimated using Kaplan-Meier and Cox) were survivals (S) without transplantation (LT), without liver related death (LRD), LRC, primary liver cancer (HCC), ascites (A), jaundice (J), encephalopathy (E), and variceal bleeding (VB). Pts with non-reliable FT (1.8%) and non-reliable TE (18%; P=0<0.001 vs FT) were excluded. Results A total of 2485 pts HCV-RNA+ with FT and TE have been included, male 56%, Caucasian 71%, SubSaharan 13%, North-African 12%, Asian 1 8%, HIV 15%; at baseline median age 50yr old; genotype 1 62%, genotype 3 8%; 7% declared >30g alcohol/day, 1 7% BMI >27kg/M2, diabetes 9%; 56% METAVIR F0F1, 25%F2F3, 19%F4. During the 7 years median-follow-up 57% were treated [402 (16%) were SVR, 1006 (41%) non-responders (NR)] and 1077 (43%) non-treated (NT). A total of 157 pts (at 10 years 20.1% [95%CI(15.0-25.3)] died, 109 from LRD (15.2%[10.2-20.3]); 31 patients (1.5% [1.0-2.1 ] at 10 years) have been transplanted; the incidence of first events occurring at least 6 months after FT were: 54 LPC [7.0%(4.0-10.1)] including 43 HCC [6.7%(1.2-10.3), 13 A [1.5(0.4-2.6)] and 2 E (0.1%). Baseline FT and TE had significant and not different performances for OS, S-LT, S-LRD and S-HCC. Only FT had significant performances for predicting the rare occurrence of A and VB (Table). Conclusion: In pts with CHC, FT and TE had highly significant performances at 10 years for predicting OS, S-LT and HCC. The advantage of FT was the higher reliability rate and a possible higher sensitivity for predicting ascites and variceal bleeding.

Survival endpointFibroTest n=2485TE n=2485FibroTest vs TE n=2485
 Risk Ratio (95%CI) P-Value adjusted on treatment, virological response, center HIV, age and gender% AUROC P-Value
Overall110 (51-239) =.000420 (13-31)=.000180 (76-83) vs 80 (75-84) =.97
Without transplantation189 (78-455) =.000129 (18-45) =.000181 (77-85) vs 80 (76-84) =.57
Without liver related death441 (149-1000) =.000149 (29-84) =.000180 (75-84) vs 81 (76-85) =.23
Without liver complications146 (71-301) =.00011.9 (0.3-12) =0.5185 (81-89) vs85 (79-89) =.91
Without HCC>500 (276-1000) =.000135 (14-89) =.000187 (83-91) vs 88 (83-91) =.95
Without ascites>500 (87-1000) =.0002Cox not applicable90(84-94) vs 91 (78-97) =0.57
Without bleeding>500 (14-1000) =.003Cox not applicable84 (60-94) vs 87 (53-97)=0.45

Disclosures:

Thierry Poynard-Advisory Committees or Review Panels: Merck; Grant/Research Support: BMS, Gilead; Stock Shareholder: Biopredictive

Yen Ngo - Employment: BioPredictive

Juliette Foucher - Board Membership: roche; Speaking and Teaching: BMS, MSD, Gilead

Mona Munteanu - Employment: Biopredictive

Olivier Deckmyn - Management Position: BioPredictive; Stock Shareholder: Bio-Predictive

Pascal Lebray - Grant/Research Support: Schering Plough; Speaking and Teaching: Janssen, MSD, Gilead

Vlad Ratziu - Advisory Committees or Review Panels: GalMed, Abbott, Genfit, Enterome, Gilead; Consulting: Astellas, Axcan, Pfizer, Sanofi-Synthelabo, Genentech, Nycomed

Victor de Ledinghen - Advisory Committees or Review Panels: Merck, Janssen, Gilead, Echosens, Boehringer Ingelheim, Abbvie; Grant/Research Support: Roche, Gilead, Janssen; Speaking and Teaching: Roche, Echosens

The following people have nothing to disclose: Julien Vergniol, Wassil Merrouche, Vincent Thibault

1435

Study of the influence of HLA genetic factors on the vertical transmission and chronification of hepatitis C virus

Angeles Ruiz-Extremera1,2, Esther-José Pavón-Castillero3, Monica Florido3, Paloma Muñoz-de-Rueda2,3, Jose Antonio Muñoz-Gámez3, Jorge Casado3, Angel Carazo3, Rosa Quiles2,3, Laura Sanjuan4, Sergio Manuel Jiménez-Ruiz4, Josefa León2,3, Javier Salmeron2,3;

1Pediatric Unit, San Cecilio University Hospital, Granada, Spain; 2CIBER Enfermedades Hepáticas y Digestivas, CIBERehd, Instituto de Salud Carlos III, Granada, Spain; 3Clinical Management Unit of Digestive Diseases, UNAI, San Cecilio University Hospital, Granada, Spain; 4Medicine Department, Granada University, Granada, Spain

BACKGROUND: Vertical transmission (VT) is considered the main route of hepatitis C virus (HCV) infection in children but the risk factors in this respect have yet to be fully identified, except maternal HIV co-infection. The aim of this study is to analyze the role of human leukocyte antigen (HLA) genetic factors in HCV-VT. PATIENTS AND METHODS: Between September 1991 and December 2009, 123 HCV-RNA positive (HIV negative) mothers were recruited, with their 1 30 children. The following risk factors for HCV were analyzed: HCV viral load, viral genotype, IL28B polymorphism (single nucleotide polymorphism rs12979860), delivery mode, ALT levels and breastfeeding. The infants were tested for HCV-RNA at birth, at 2, 4, 6, 8, 10, 12, 18 and 24 months, and then annually at 3, 4, 5 and 6 years. VT was defined as children who presented HCV-RNA positive for at least two subsequent blood samples. Chronic or persistent infection was defined as children with HCV-RNA positive at the end of the study. The frequencies of HLA class I alleles (A, B and Cw) and of HLA class II alleles (DRB1, DQA1, DQB1, DPA1 and DPB1) were analyzed. RESULTS: Of the 123 mothers (and 130 children) included in the study, VT occurred in 24 cases. Of these, 8 children suffered chronic infection and in 1 6 the virus was eliminated. VT study: Among the 123 mothers, the alleles related to VT were the presence of B*0702 (P=0.024), Cw*0702 (P=0.050) DQA1*0301 (P=0.011, Pc=0.04) and DPB1*0201 (P=0.046) and the absence of B*3501 (P=0.030) and Cw*0602 (P=0.006, Pc=0.05). Among the 130 children, however, the only allele related to VT was the presence of DPB1*0201 (P=0.038). Chronification of the virus: In this case, the associated alleles of the mothers (n=24) were DQA1 *0101 (P=0.028) and DQB1*0604 (P=0.028), and among the children (n=24) they were B*0801 (P=0.037), DRB1*0301 (P=0.037), DQA1*0501 (P=0.037) and DQB1*0603 (P=0.034). Mother/child allelic concordance was higher among the children with chronic infection than in those with no chronification (67% ± 4.06 vs. 57% ± 1.34, P=0.045). CONCLUSIONS: The HLA alleles of the mother are of greater importance than those of the child with respect to HCV-VT. Thus, the presence of the HLA allele, class I Cw*0602 and of the HLA allele, class II DQA1 *0301 in the mother is directly related to VT. Moreover, the greater allelic concordance among the children, with respect to their mothers, is associated with the chronification of the virus in these children.

Disclosures:

The following people have nothing to disclose: Angeles Ruiz-Extremera, Esther-José Pavón-Castillero, Monica Florido, Paloma Muñoz-de-Rueda, Jose Antonio Muñoz-Gámez, Jorge Casado, Angel Carazo, Rosa Quiles, Laura Sanjuan, Sergio Manuel Jiménez-Ruiz, Josefa León, Javier Salmeron

1436

Clinical performance of rapid diagnostic tests for broad-scale HCV screening

Stephane Chevaliez1, Lila Poiteau1, Isabelle Rosa2, Alexandre Soulier1, Francoise Roudot-Thoraval3, Christophe Hezode4, Jean-Michel Pawlotsky1;

1Virology & INSERM UnitU955, HenriMondor University Hospital, Creteil, France; 2Hepatology and Gastroen-terology, Centre Hospitalier Intercommunal de Creteil, Creteil, France; 3Public Health & INSERM Unit U955, University Hospital Henri Mondor, Creteil, France; 4Hepatology & INSERM Unit U955, University Hospital Henri Mondor, Creteil, France

Background: Large-scale HCV screening is mandatory in order to prevent further spread of the infection, improve access to care in the context of new HCV drug development, and reduce the risk of long-term complications. Rapid diagnostic tests (RDTs) represent an attractive alternative to methods based on enzyme immunoassay (EIA) from whole blood samples collected by venous puncture, because they use original matrix specimens (such as fingerstick capillary whole blood or oral fluid) and offer the advantages of simplicity, limited need for instrumentation, minimal training required, and rapid performance at room temperature. Objective: The aim of this study was to assess the clinical performance of different RDTs for the detection of total anti-HCV antibodies (anti-HCV Ab) in fingerstick capillary whole blood and in oral fluid. Patients & Methods: 420 individuals (54% male, mean age 51.1 ± 15.2 years) were included; 159 were HCV-seronegative and 261 were HCV-seropositive [treatment-naïve in 91.9% of cases, quantifiable HCV RNA in 92.0% (5.7±1.0 Log10 IU/mL), infected with HCV genotype 1 in 63.0% of cases]. Four RDTs were performed in each individual: 3 from fingerstick capillary whole blood [OraQuick® HCV Rapid Antibody test (Orasure), Toyo® anti-HCV (Turklab) and Labmen HCV test (Turklab)] and the OraQuick® HCV Rapid Antibody test from oral fluid. Results: The RDT result was interpretable in more than 99% of 1,260 tests performed. The specificity in fingerstick capillary whole blood varied from 98% to 100% according to the test. The clinical sensitivity, tested in the 261 HCV-seropositive patients, was 99.6% (95%CI: 97.9-99.9%), 96.4% (95%CI: 93.3-98.1%) and 63.4% (95%CI: 53.3-72.5%) for OraQuick®, Toyo® and Labmen RDTs, respectively. Anti-HCV Ab were not detected by the OraQuick® test in only 1 of 261 patients (this patient was infected with genotype 1b, HCV RNA was 6.0 Log10 IU/mL), whereas 9 patients were falsely negative with the Toyo® test, including 8 with detectable/quantifiable HCV RNA. The clinical sensitivity of OraQuick® HCV Rapid Antibody test for oral fluid was satisfactory [98.0% (95%IC: 95.5-99.2%)]. Anti-HCV Ab were not detected in only 5 HCV-seropositive subjects, who had detectable anti-HCV Ab in oral fluid by 3rd-generation EIA in 5 cases. Conclusion: The rapid diagnostic HCV tests evaluated here were easy to perform, rapid and specific, with an advantage for the OraQuick test. RDT in fingerstick capillary whole blood or oral fluid thus appears as a promising new tool for broad-scale screening of HCV infection in high- to medium-risk populations.

Disclosures:

Francoise Roudot-Thoraval - Advisory Committees or Review Panels: Roche, Roche; Consulting: LFB; Speaking and Teaching: Gilead, Gilead, Roche

Christophe Hezode - Speaking and Teaching: Roche, BMS, MSD, Janssen, abbvie, Gilead

Jean-Michel Pawlotsky- Consulting: Abbott, Achillion, Boehringer-Ingelheim, Bristol-Myers Squibb, Idenix, Gilead, Janssen, Madaus-Rottapharm, Merck, Novartis, Roche; Grant/Research Support: Gilead; Speaking and Teaching: Boehringer-Ingelheim, Bristol-Myers Squibb, Gilead, Madaus-Rottapharm, Merck, Janssen-Cilag, Novartis, Abbott

The following people have nothing to disclose: Stephane Chevaliez, Lila Poiteau, Isabelle Rosa, Alexandre Soulier

1437

A rs4374383 Single Nucleotide Polymorphisms of MERTK gene is linked to a higher likelihood of hepato-cellular carcinoma in patients with HCV cirrhosis

Vito Di Marco1, Vincenza Calvaruso1, Stefania Grimaudo1, Donatella Ferraro2, Maria Grazia Bavetta1, Antonietta Di Cristina1, Giuseppe Cabibbo1, Elisabetta Conte1, Antonio Craxi1;

1Sezione di Gastroenterologia & Epatologia, DIBIMIS, University of Palermo, Palermo, Italy; 2Servizio di Virologia, Dipartimento di Igiene “G.D'Alessandro”,, University of Palermo, Palermo, Italy

Background and Aims: MERTK gene, located in chromosome 2, encode factors involved in phagocytosis of apoptotic cells by macrophages. GWA studies reported that the rs4374383 SNP of MERTK gene is associated with the risk of developing fibro-sis in patients with HCV chronic hepatitis. We evaluated if rs4374383 SNP influenced the risk of liver decompensation (LD) and hepatocellular carcinoma (HCC) in patients with HCV cirrhosis undergoing antiviral therapy. Methods: In a prospective cohort of patients with compensated HCV cirrhosis treated with Peg-interferon alfa-2b and ribavirin (P/R), rs4374383 SNP was carried out using the TaqMan SNP genotyping allelic discrimination method (Applied Biosystems, CA, USA) on sera stored before treatment. All patients were screened for esophageal varices (EV) before treatment and underwent surveillance for HCC every six months. Univariate and multivariate Cox regression analysis was used to determine factors associated with development of LD and HCC. Results: Among 349 patients (mean age 58±8.6 years, 61.2% men, 85% genotype 1) included in this analysis, 16.9% had AA genotype, 46.4% GA genotype and 36.9% GG genotype of rs4374383 SNP, and 50.7% had EV at baseline. Eighty-seven patients (24.9%) achieved a Sustained Virological Response (SVR). During follow-up (median 77 months; range 12-145) 6 (6.8%) SVR and in 71 (27.1%) no SVR patients developed LD (p<0.001), while 6 patients (6.8%) with SVR and 66 patients (25.2%) without SVR developed HCC (p<0.001). By multivariate analysis EV (HR 3.11; 95%CI 1.69-5.75; p<0.001), platelet count (HR 0.99; 95%CI 0.98-0.99; p=0.001), albumin (HR: 3.11; 95%CI 0.19-0.54; p<0.001), and absence of SVR (HR: 4.04; 95%CI 1.63 -10.05; p= 0.003) were independently associated to LD. The variables independently associated to development of HCC were age (HR 1.04; 95%CI 1.01-1.07; p=0.045), GGT (HR 1.14; 95%CI 1.20-1.37, p=0.008), absence of SVR (HR 3.31; 95%CI 1.43-7.68; p=0.005) and the AA genotype of rs4374383 SNP (HR 2.67; 95% CI 1.36 -5.23; p= 0.004). The risk of developing HCC was of 1.04 per 100 persons/years in patients with SVR, and of 2.43, 4.05 and 7.17 per 100 persons/years in non responders to therapy with genotype GG, GA and AA of rs4374383 SNP, respectively. Conclusion: The AA allele of rs4374383 SNP of MERTK gene is associated with a higher risk of developing HCC in patients with HCV cirrhosis not responding to P/R. Since the MERTK gene is a regulator of tumor-associated macrophages involved in the modulation of inflammatory responses and in angiogenesis, its polymorphism could affect the rate of development of HCC in a predisposed population.

Disclosures:

The following people have nothing to disclose: Vito Di Marco, Vincenza Cal-varuso, Stefania Grimaudo, Donatella Ferraro, Maria Grazia Bavetta, Antonietta Di Cristina, Giuseppe Cabibbo, Elisabetta Conte, Antonio Craxi

1438

Significant roles of IFN-λ3 with reciprocal levels in serum and ex vivo stimulated PBMC in chronic hepatitis C

Yoshihiko Aoki1, Kazumoto Murata1, Masaya Sugiyama1, Tatsuji Kimura2, Tsutomu Takeda1, Sachiyo Yoshio1, Nao Nishida1, Yoko Yamagiwa1, Masaaki Korenaga1, Masatoshi Imamura1, Tatsuya Kanto1, Naohiko Masaki1, Jong-Hon Kang3, Masashi Mizokami1;

1The Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine, Tokyo, Japan; 2Insutitute of Immunology Co., Ltd, Tokyo, Japan; 3Center for Gastroenterology, Teine-Keijinkai Hospital, Sapporo, Japan

Background&Aims: Genetic variation in IL28B has been found as a predictive factor for pegylated-interferon/ribavirin (Peg-IFN/RBV) therapy in chronic hepatitis C (CHC) patients. We recently reported that BDCA3+dendritic cell (DC) or BDCA4+DC robustly produce IFN-λ3 by stimulation with toll-like receptor (TLR)3 or TLR7 agonist, respectively, suggesting IFN-λ3 production is closely related to innate immunity. We also found that high levels of ex vivo induced IFN-λ3 by TLR7 agonist correlated with the favorable response to Peg-IFN/RBV therapy. Innate immunity could play a mechanistic role in other viral hepatitis. Methods: We collected serum samples from patients with acute hepatitis due to hepatitis A virus (HAV), hepatitis E virus (HEV), Epstein-Bar virus (EBV) and chronic hepatitis due to hepatitis B virus (HBV), hepatitis C virus (HCV). Serum form healthy volunteers (n=24) were used as control. Serum levels of IFN-λ3, using frozen stocks, were measured by our newly developed chemiluminescence enzyme immunoassays (CLEIA). No treatment was given when serum samples were taken in all patients. In CHC patients, PBMC was collected on the same day as serum was taken. Purified PBMC was stimulated with IFN-α for 16 h. After stimulation with TLR7 agonist for 24 h, the supernatant was subjected to CLEIA. SNP near IL28B (rs809991 7; TT is a favorable genotype for Peg-IFN/RBV therapy) were evaluated by InvaderPlus assay in CHC patients.Results: Serum IFN-λ3 levels were significantly higher in all patients than those in healthy volunteers (n = 24: 1.4 ± 0.9). Among them, serum IFN-λ3 levels in HCV (n = 87: 23.5 ± 28.7), HEV (n = 9: 22.4 ± 19.0) or HAV (n = 5: 12.0 ± 12.3) were significantly higher than those in HBV (n = 21: 4.9 ±5.1) or EBV (n = 5: 3.2 ± 2.0). In all patients with acute hepatitis A or E, serum IFN-λ3 levels were returned to the similar levels to healthy volunteers after recovery of diseases. These results suggest that RNA viruses, especially HCV, are more apt to induce IFN-λ3 than DNA viruses. Next, we focused on CHC patients and examined the association between ex vivo induced IFN-λ3 levels and serum IFN-λ3 levels. No significant differences in serum and ex vivo induced IFN-λ3 levels were observed between IL28B genotype. Interestingly, high levels of ex vivo induced IFN-λ3 were significantly observed in patients with low levels of serum IFN-λ3 (p = 0.035). Conclusion: IFN-λ3 may play a major role in RNA viruses-related liver diseases. Capacities for IFN-λ3 production in PBMC were reciprocally correlated with serum IFN-λ3 levels, which may contribute to address the mechanistic roles of IFN-λ3 in CHC patients.

Disclosures:

Tatsuji Kimura - Employment: Institute of Immunology, Co., Ltd.

The following people have nothing to disclose: Yoshihiko Aoki, Kazumoto Murata, Masaya Sugiyama, Tsutomu Takeda, Sachiyo Yoshio, Nao Nishida, Yoko Yamagiwa, Masaaki Korenaga, Masatoshi Imamura, Tatsuya Kanto, Naohiko Masaki, Jong-Hon Kang, Masashi Mizokami

1439

Presence of advanced liver disease concurrent with initial diagnosis of hepatitis C virus (HCV) infection among patients in the Chronic Hepatitis Cohort Study (CHeCS): missed opportunities for intervention

Anne C. Moorman1, Jian Xing1, Loralee B. Rupp2, FujieXu1, Stuart C. Gordon2, Mei Lu2, Philip R. Spradling1, Eyasu H. Teshale1, Joseph A. Boscarino3, Vinutha Vijayadeva5, Mark A. Schmidt4, Scott D. Holmberg1;

1Division of Viral Hepatitis, National Center for HIV, Viral Hepatitis, STD, and TB Prevention, Centers for Disease Control and Prevention, Atlanta, GA; 2Henry Ford Health System, Detroit, MI; 3Geisinger Health System, Danville, PA; 4The Center for Health Research, Kaiser Permanente Northwest, Portland, OR; 5Kaiser Permanente Hawaii, Honolulu, HI

Background. Because HCV infection is asymptomatic until cirrhotic decompensation occurs, screening and treatment of asymptomatic persons is needed to avert progression to advanced fibrosis. To determine the frequency of advanced liver disease at the time of initial diagnosis of HCV infection, we analyzed data from CHeCS, a large observational cohort study of persons with chronic viral hepatitis who receive care at four US healthcare systems. Methods. The date of “initial” HCV diagnosis was defined as the earliest of: electronic health record or clinic chart report of a positive laboratory test, an HCV-related diagnostic or procedure code, or patient report via survey. Our analyses were restricted to patients who had their initial HCV diagnosis ≥ 6 months after their first encounter with the health system, and who had ≥ 12 months further observation. We determined the proportion of patients with an initial HCV infection diagnosis concurrent with (i.e., 3 months before or up to 12 months after) hepatic fibrosis, defined as a liver biopsy indicating cirrhosis or mean FIB-4 score >5.88. We also determined the proportion of patients with diagnoses indicating severe liver disease (ICD9 or procedure codes indicating liver transplant, hepatocellular carcinoma, liver failure, hepatic encephalopathy, portal hypertension, esophageal varices, other gastroesophageal hemorrhage, ascites, and other sequelae of chronic liver disease) by proximity to time of initial HCV diagnosis. Results. Of 12,529 patients with ≥ 1 visit to a CHeCS clinic during 2006-2010, 6,262 (50%) met the inclusion criteria for observation around the time of initial HCV diagnosis. Of these, 701 (11%) had severe hepatic fibrosis concurrent with initial HCV diagnosis, and similarly 712(11%) had their first severe liver disease diagnosis either prior to or within 12 months of their initial HCV diagnosis. An additional 545 (9%) had a severe liver disease diagnosis within 1 -5 years, and 383 (6%) had such a diagnosis >5 years after their initial HCV diagnosis. Conclusions. A sizeable minority of CHeCS patients had advanced liver disease concurrent with their initial HCV diagnosis. These findings suggest missed opportunities for diagnosis and therapeutic intervention before the onset of severe liver disease when treatment may involve high cost and diminished outcomes.

Disclosures:

Stuart C. Gordon - Advisory Committees or Review Panels: Tibotec; Consulting: Merck, CVS Caremark, Gilead Sciences, BMS; Grant/Research Support: Roche/Genentech, Merck, Vertex Pharmaceuticals, Gilead Sciences, BMS, Abbott, Intercept Pharmaceuticals, Exalenz Sciences, Inc.

The following people have nothing to disclose: Anne C. Moorman, Jian Xing, Loralee B. Rupp, Fujie Xu, Mei Lu, Philip R. Spradling, Eyasu H. Teshale, Joseph A. Boscarino, Vinutha Vijayadeva, Mark A. Schmidt, Scott D. Holmberg

1440

Population-Based Genome-Wide Association Study to Identify Potential Single Nucleotide Polymorphisms Associated with the Serum Levels of Alanine Aminotransferase among Asymptomatic Chronic Hepatitis C Patients

Mei-Hsuan Lee1, Hwai-I Yang2,3, Sheng-Nan Lu4, Yu-Ju Lin2,5, Pao-Jen Liu6, Yu-Chuan Chien6, Chin-Lan Jen2, San-Lin You2, Li-Yu Wang7, Yong Yuan8, Gilbert L'Italien8,9, Chien-Jen Chen2,10;

1Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan; 2Genomics Research Center, Academia Sinica, Taipei, Taiwan; 3Graduate Institute of Clinical Medical Science, China Medical University, Taichung, Taiwan; 4Department of Gastroenterology, Kaohsiung Chang-Gung Memorial Hospital, Kaohsiung, Taiwan; 5Graduate Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan; 6Mol-ecular and Genomic Epidemiology Center, China Medical University Hospital, Taichung, Taiwan; 7McKay Medical College, Taipei, Taiwan; 8Global Health Economics and Outcome Research, Bristol-Myer Squibb, Princeton, NJ; 9Yale University School of Medicine, New Haven, CT; 10Graduate Institute of Epidemiology, College of Public Health, National Taiwan University,, Taipei, Taiwan

Background & Aims: Serum levels of alanine aminotransferase (ALT) test is widely used in clinical settings for diagnosis of liver diseases and monitoring for hepatitis C patients. In addition, this seromarker is presented as an inflammatory response marker against hepatitis C virus (HCV) infection, and the elevated serum levels of ALT increased the risk for hepatocellular carcinoma among chronic hepatitis C patients. We performed a genome-wide association study to discover single nucleotide polymorphisms (SNPs) associated with the serum levels of ALT among chronic hepatitis C patients without treatment experience. Methods: A total of 808 anti-HCV-seropositive and HBsAg-seronegative participants were recruited during 1991-1992. All of them were free of hepatocellular carcinoma cases during the follow-up year of 1991-2008. The serum samples were collected at the enrollment for the test of serum levels of ALT. We applied the AxiomTM Genome-Wide CHB Array, a recently developed tool specifically on Chinese Han population that provides maximum power for GWAS and has capability for genomic researchers to identify trait-associated SNPs in the Han Chinese. The serum levels of ALT was served as a quantitative trait in the analyses to test the association for each SNP. The significant p value was set as 8.1 ×l O-8 by Bonferroni correction. The log10 transformed serum levels of ALT by various genotypes at each SNP were tested by t-test or ANOVA test. The logistic regressions were used to estimate odds ratios (ORs) and 95% confidence intervals (95% CIs) of the potential SNPs associated with serum levels of ALT. Results: There were 372 (46.0%) with serum levels of ALT&LE;15 U/L, 320 (39.6%) 16-45 U/L, and 116 (14.4%) >45 U/L among the asymptomatic HCV infected participants. The means and standard deviation of the serum levels of ALT were 28.3±37.6. In total, 613,774 SNPs with call rate >95%, minor allele frequency >0.01, were included in the analyses. We found 6 SNPs potentially associated with the serum levels of ALT. These SNPs were located on the chromosome 2,5,7,1 0 and 21. The mean values of serum levels of ALT had significant differences by various genotypes at each SNP (p<0.001). The serum levels of ALT was categorized as normal and abnormal with the cut-off of 45 U/L. The odds ratios for the SNPs ranged from 1.8 to 3.1 associated with abnormal serum levels of ALT. Conclusion: There were SNPs identified to be potentially associated with serum levels of ALT, a seromarker of inflammatory response, in chronic hepatitis C patients. However, these SNPs should be validated by an independent external population and functional studies would be needed.

Disclosures:

Yong Yuan - Employment: Bristol Myers Squibb Company

Gilbert L'Italien - Employment: Bristol Myers Squibb; Stock Shareholder: Bristol Myers Squibb

The following people have nothing to disclose: Mei-Hsuan Lee, Hwai-I Yang, Sheng-Nan Lu, Yu-Ju Lin, Pao-Jen Liu, Yu-Chuan Chien, Chin-Lan Jen, San-Lin You, Li-Yu Wang, Chien-Jen Chen

1441

HCV Antibody Dynamics following Clearance of acute HCV Infection and subsequent Reinfection among HIV-infected Men who have Sex with Men

Joost W. Vanhommerig1, Maria Prins2,4, Xiomara V. Thomas1, Richard Molenkamp1, Jan T. van der Meer3, Sylvia M. Bruisten4, Janke Schinkel1;

1Medical Microbiology, Academic Medical Center, Amsterdam, Netherlands; 2Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, Amsterdam, Netherlands; 3Internal Medicine, Academic Medical Center, Amsterdam, Netherlands; 4Infectious Diseases, Public Health Service, Amsterdam, Netherlands

Introduction. HCV antibody tests (anti-HCV) are commonly used as a qualitative measure for past or present HCV infection and are not used for detection of HCV reinfection, because one will remain antibody positive after exposure. Recently, a high incidence of HCV reinfection (15.2/100 person-years) was reported after treatment-induced clearance among HIV-infected men who have sex with men (MSM). We investigated dynamics of anti-HCV reactivity and the ability to detect HCV reinfection by monitoring anti-HCV, compared to testing RNA and/or ALT concentrations. Methods. N=63 HIV-1-infected MSM with acute HCV infection were included. Acute infection was defined as having an interval of max. 6 months between the last negative and the first positive HCV RNA test. Presence of HCV RNA was determined by TMA Versant (Siemens) or CAP/CTM (Roche). Anti-HCV reactivity, indicated by S/CO ratio, was measured every 7.0 months (median, IQR 3.8-1 1.7) using the AxSYM HCV v3.0 assay (Abbott). Concentrations of ALT were measured every 1.5 months (median, IQR 0.7-3.6) and considered to be elevated when above the upper limit of the normal range (ULN), <40 U/L. To distinguish between relapse and reinfection with the same genotype, E2/HVR1 sequences were analyzed before and after treatment. Results. Among 63 MSM, 4 spontaneously cleared and 43 initiated HCV treatment during the acute phase of infection. This resulted in a sustained viro-logic response (SVR) in 31/43 (72%). At the first RNA positive date during primary HCV infection, ALT was elevated in 85% of cases, and anti-HCV was reactive in 35% of cases. During a median follow-up of 2 years after clearance of HCV, a significant decline in anti-HCV reactivity was observed. Peak and subsequent nadir anti-HCV S/CO ratios were 72.5 (median, IQR 50.2-110.6) and 3.62 (median, IQR 1.2-27.8), respectively. Following reinfection (N = 11), anti-HCV reactivity increased notably in all cases, to an S/CO ratio of 83.7 (median, IQR 76.4-146.7). ALT peak levels following reinfection were less pronounced, and were elevated at the first RNA positive date in 63% of cases. Conclusions. A relatively fast decline of anti-HCV reactivity was documented after clearance of HCV. Following reinfection, anti-HCV responses were comparable to responses after primary infection. ALT is a good marker for primary HCV infection, however this may not be the case for reinfection. Given the lower anti-HCV test frequency in this study, monitoring anti-HCV reactivity may be a cost-effective approach for diagnosis of HCV reinfection after treatment-induced or spontaneous clearance.

Disclosures:

Maria Prins - Speaking and Teaching: msd, roche

Richard Molenkamp - Independent Contractor: Roche Diagnostics

The following people have nothing to disclose: Joost W. Vanhommerig, Xiomara V. Thomas, Jan T. van der Meer, Sylvia M. Bruisten, Janke Schinkel

1442

Stagnation of histological improvement is a predictor of the risk of hepatocellular carcinoma after eradication of hepatitis C virus

Akihiro Tamori1, Shoji Kubo2, Sawako K. Uchida1, Atsushi Hagi-hara1, Etsushi Kawamura1, Hideki Fujii1, Shuji Iwai1, Hiroyasu Morikawa1, Masaru Enomoto1, Yoshiki Murakami1, Norifumi Kawada1;

1Hepatology, Osaka City University Graduate School of Medicine, Osaka, Japan; 2Department of Hepato-Biliary-Pancre-atic Surgery, Osaka City University, Osaka, Japan

Objectives: In the near future, a sustained viral response (SVR) will be achieved in more than 90% of patients with hepatitis C virus (HCV) by treatment with direct-acting antiviral agents. On the other hand, it is well known that hepatocellular carcinoma (HCC) develops after eradication of HCV. The aim of this study was to delineate the clinical characteristics of patients in whom HCC develops after attaining an SVR. Patients and methods: 129 patients with SVR (71 males, 58 females) followed-up for more than 4 years after interferon-based anti-HCV therapy were studied. Ultrasonography or contrasted computed tomography was performed every 6 months (observation period, 48-248 months). HCC developed in 1 8 patients (1 6 males, 2 females). We compared clinical and histological factors between the HCC group and non-HCC group. Liver biopsy was performed twice in 14 patients in whom HCC developed and 31 patients without HCC; the first biopsy was performed at the beginning of interferon therapy, and the second biopsy was done more than 3 years after the final interferon injection. Histological changes were also compared between the two groups. Results: HCC predominantly developed in males with SVR (p = 0.004). Age at interferon treatment, body mass index, presence of diabetes mellitus, and HCV genotype did not differ significantly between the HCC group and non-HCC group. However, the rate of anti-HBc positivity and the levels of alanine aminotransferase and alpha-fetoprotein before interferon therapy were significantly higher and the platelet count was significantly lower in the HCC group. Before anti-HCV therapy, the histological stage of fibrosis was respectively 1-2 and 3-4 in 87 patients and 13 patients in the non-HCC group, as compared with 10 and 7 patients in the HCC group. We assessed the improvement in fibrosis stage by comparing the findings of the first and second biopsies. The improvement in the fibrosis stage per year was 0.036 in the HCC group and 0.228 in the non-HCC group (p = 0.01). Conclusions: Risk factors for the development of HCC in patients with SVR were male sex, advanced fibrosis before interferon therapy, and stagnation of histological improvement after HCV eradication.

Histological findings between non-HCC group and HCC group

 Patietns without HCCPatients with HCCp-value
 n= 100n=17 
Histological staging 1/2/3/466/21/10/30/10/6/10.012
 n = 31n=14 
Improvement of fibrosis stage0.228 per year0.036 per year0.01

Disclosures:

Akihiro Tamori - Grant/Research Support: MSD

The following people have nothing to disclose: Shoji Kubo, Sawako K. Uchida, Atsushi Hagihara, Etsushi Kawamura, Hideki Fujii, Shuji Iwai, Hiroyasu Morikawa, Masaru Enomoto, Yoshiki Murakami, Norifumi Kawada

1443

Diagnosis and Treatment of HCV-related mixed cryo- globulinemia over the last decade at a Liver Referral Center

Yumi Ando1, Peter D. Gorevic2, Thomas D. Schiano3;

1Department of Medicine, Mount Sinai Medical Center, New York, NY;2Depart-ment of Medicine-Rheumatology, Mount Sinai Medical Center, New York, NY; 3The Recanati/Miller Transplantation Institute and the Division of Liver Diseases, Mount Sinai Medical Center, New York, NY

Background: HCV-related mixed cryoglobulinemia (MC) is an uncommon extrahepatic manifestation of HCV. Patients infected with HCV commonly have the asymptomatic presence of cryoglobulins and rheumatoid factor. Clinically significant MC is however less commonly seen. Typical manifestations of this disorder include cutaneous purpura, peripheral neuropathy, arthralgia, and glomerulonephritis (GN). In this study, in order to better characterize the illness, we sought to identify patients with HCV having clinically significant MC based on clinical history and pathologic findings.. Methods: We performed a retrospective chart review of patients with both HCV and clinically significant MC seen at Mount Sinai Medical Center from 2003-2013. These patients were identified using the Mount Sinai Data Warehouse software by mining all relevant patient care computer systems. The inclusion criteria were defined as those with HCV who tested positive for cryoglobulins and also had manifestations of vasculitis, GN, peripheral neuropathy, and/or arthropathy not explained by other underlying disorders. Results: 51 patients were identified as having HCV and clinically significant MC. Male to female ratio was 1:1 (49% M, 51% F) and the most common HCV genotype was type 1 (59%). Cirrhosis was present in 28 (55%). Of the manifestations of MC, cutaneous vasculitis was the most common (35%), followed by GN (25%). There was significant overlap among the manifestations and 14 (27%) had both cutaneous vasculitis and GN. Of those with GN, 13 (48%) progressed to ESRD requiring hemodialysis. 18 patients had manifestations severe enough to warrant plasmapheresis, and 11 patients were treated with rituximab. Of those who received rituximab, 10 (91%) had GN. Only 3 (30%) with GN treated with rituximab progressed to ESRD versus 10 (59%) with GN who did not receive rituximab. Overall, MELD progression (difference in MELD between most recent and initial visit) in patients treated with rituximab was on average 2.3 points versus 5.5 points in those who did not get rituximab. MELD progression on average was 5.3 for those who received some form of antiviral therapy and only 3.1 in those who were treatment naive. Conclusion: Our study suggests that rituximab may be effective in preventing progression of HCV-related MC, especially those with GN. This is especially important because renal insufficiency often precludes patients from treatment with standard antiviral regimens.

Disclosures:

Thomas D. Schiano-Advisory Committees or Review Panels: vertex, salix, merck, gilead, pfizer; Grant/Research Support: massbiologics, itherx

The following people have nothing to disclose: Yumi Ando, Peter D. Gorevic

1444

IP 10, IFI 27, MX1 and ISG 15 genes expression in liver specimens and IL-28B SNPs rs12979860, rs8099917, rs 12980275 in patients with chronic hepatitis C

Krzysztof Domagalski1 ,2, Malgorzata Pawlowska1, Andrzej Tre-tyn2, Waldemar Halota';

1Department of Infectious Diseases&Hepatology Collegium Medicum N.Copernicus University, Bydgoszcz, Poland; 2Department of Plant Physiology and Biotechnology, Faculty of Biology and Environmental Protection, Nicolaus Copernicus University, Torun, Poland

Aim: Assessment of gene IP10, IFI27, MX1 i ISG15 expression in liver specimens and determination of its relationship with genotype markers within the IL-28B SNP (rs12979860, rs8099917, rs12980275), including the results of histological evaluation of patients with chronic viral hepatitis C. Material and Methods: In liver specimens taken from 64 patients with chronic hepatitis C molecular analysis of DNA and RNA and histopathological examination were performed. Analysis of polymorphisms rs1 2979860, rs8099917 and rs12980275 was performed with PCR-RFLP techniques using restriction enzymes BstUI, BseMI, BseLI, respectively. To assess the relative expression level of IP10, IFI27, MX1 and ISG15 real-time PCR method was used. GAPDH was used as a reference gene. The study compared the gene expression in patients with favorable genotypes for a given marker adverse (CC vs CT-TT for rs1 2979860, TT vs TG-GG for rs8099917, and AA vs AG-GG for rs12980275). Histopathological evaluation of liver specimens was performed on the basis of the Scheuer's modified scale. Results: Analysis included 64 patients (median age 38, range 19-63) mainly men (81%). 49 (77%) patients had ALT activity > 40 U/l. Distribution of fibrosis: F0 (6%), F1 (50%), F2 (37%), F3 (7%), F4 (3%) and inflammatory activity: A0 (0%), A1 (19%), A2 (69%), A3 (9%) A4 (0%). The distribution of genotypes for marker rs1 2979860 was CC 40.6%, CT 45.3%, TT 14.1%, for rs8099917TT 59.4%, TG 32.8, GG 7.8% and for rs1 2980275 AA 39.1 %, AG 46.8%, GG 14.1 %. The presence of favorable prognostic genotype was statistically significantly associated with decreased relative expression of genes IFI27, MX1 i ISG15 in liver specimens compared with patients with unfavorable genotypes for each marker analyzed (p<0,001). IP 10 gene was also observed for the reduction of favorable genotypes of the analyzed markers, however the differences were not statistically significant (p>0.05). Regardless of the analyzed markers the largest differences were observed in expression of gene IFI27. The comparative analysis showed that rs12979860 differentiates the most the expression of the analyzed genes. No effect of liver disease advancement was found on the observed relationship between genetic variation and gene expression. Conclusions: A relationship was found between genotype markers rs1 2979860, rs8099917, rs1 2980275 and the level of expression of ISG genes, such as IFI27, MX1 i ISG15. Patients with favorable IL-28B genotypes are characterized by a lower ISG genes expression. The effect of variation within IL-28B on the results of treatment may be associated with changes in ISG genes expression profiles induced by interferon.

Disclosures:

The following people have nothing to disclose: Krzysztof Domagalski, Malgorzata Pawlowska, Andrzej Tretyn, Waldemar Halota

1445

The effect of antiviral therapy on progression of liver disease in Hepatitis C Virus-infected patients with malignancies

Parag Mahale1, Ethan D. Miller2, Boris Blechacz2, H. Franklin Her-long2, Marta Davila2, Harrys A. Torres1;

1Infectious Diseases, Infection Control and Employee Health, The University of Texas MD Anderson Cancer Center, Houston, TX; 2Gastroenterology, Hepatology & Nutrition, The University of Texas MD Anderson Cancer Center, Houston, TX

Background: Antiviral treatment (AVT) for hepatitis C virus (HCV) infection reduces the risk of liver disease progression. However, the effect of AVT on progression of liver disease in HCV-infected cancer patients (pts) is unknown and management guidelines are lacking for this population. We aimed to study the effect of AVT on liver disease progression in HCV-infected pts with cancer. Methods: Records of HCV-infected pts with any type of cancer seen at MD Anderson Cancer Center (2008-2011) were reviewed retrospectively. Baseline characteristics were compared between pts who did or did not receive AVT. The probability of developing cirrhosis and portal hypertension (PH) was estimated by Kaplan-Meier curves. The statistical significance of the difference between treated vs. non-treated pts was determined by log-rank test. Multivariable Cox proportional hazards regression model was used to determine the association between AVT and liver disease progression. Results: Out of 642 HCV-infected cancer pts seen during the study period, 348 (54%) received AVT. Most pts had solid tumors (n=462 or 72%), mainly hepatocellular carcinoma (n=177 or 38%), other gastrointestinal tumors (n=60 or 13%) and breast (n=41 or 9%) cancer. Non-Hodgkin lymphoma (n=98 or 57%) was the most common hematological malignancy. When baseline characteristics were compared between AVT treated vs non-treated pts, no statistically significant differences were observed in baseline cirrhosis (16% vs. 18%; p=.46), PH (8% vs. 8%, p=1.0), or cancer status (34% vs. 31%, p=.73). However, treated pts were older (mean age, 59.5 yrs vs. 56.5 yrs; p<.001), predominantly Caucasians (69% vs. 61%; p=.01), had more solid tumors (78% vs. 65%; p=.001), genotype 2/3 infection (36% vs. 24%; p=.03), lower baseline ALT levels (mean, IU/ml; 58.9 vs. 60.7; p=.08), and higher baseline INR (mean, 1.2 vs. 1.1; p=.04). Unadjusted Cox proportional hazards regression analyses showed that among those who were non-cirrhotic and non-PH at baseline, the rate of progression to cirrhosis (HR, 0.31; 95% CI, 0.18-0.52; p<.001) and PH (HR, 0.26; 95% CI, 0.13-0.5; p< .001) was lower in treated group, irrespective of the treatment outcome (SVR vs. non SVR). This lowered rate of progression to cirrhosis (HR, 0.38; 95% CI, 0.16-0.93; p=.03) and PH (HR, 0.19; 95% CI, 0.05-0.66; p=.009) persisted in multivariable Cox proportional hazards regression model. Conclusions: AVT reduces the risk of liver disease progression in HCV-infected pts with any type of malignancy and should be offered to suitable candidates as recommended for non-cancer pts.

Disclosures:

Harrys A. Torres - Advisory Committees or Review Panels: Merck, Vertex, Novartis, Astellas, Pfizer, Genentech; Grant/Research Support: Merck, Vertex

The following people have nothing to disclose: Parag Mahale, Ethan D. Miller, Boris Blechacz, H. Franklin Herlong, Marta Davila

1446

HCV Research UK - a UK national resource to support global research into HCV infection

John McLauchlan1, Will Irving2, John F. Dillon4, Graham R. Foster3, Sharon Hutchinson5;

1University of Glasgow, Glasgow, United Kingdom; 2University of Nottingham, Nottingham, United Kingdom; 3University of London, London, United Kingdom; 4University of Dundee, Dundee, United Kingdom;5University of Glasgow Caledonian, Glasgow, United Kingdom

Background: Hepatitis C virus (HCV) infection is a priority area for research and development to meet the clinical challenges posed by the scale of the infection in the UK and world-wide. A collaborative group of basic and clinical scientists within the UK set out to create a national cohort of HCV-infected patients, with a purpose-built clinical database and biorepository to act as a resource to underpin future research into all aspects of HCV infection. Aims: Our objective has been to create a multi-disciplinary consortium comprising clinicians and non-clinical scientists to encourage translational research into the factors that determine outcome of infection, treatment response and disease progression. Our specific aims are: (1) To establish a cohort of 1 0,000 patients with HCV infection from across the UK (2) To construct a bespoke clinical database and biorepository (3) To invite applications to utilise the HCV Research UK resource from all-comers including academics and industry, within and outside the UK Progress: We are recruiting patients from over 30 participating centres in the UK at a rate of around 100 per week. There are currently over 4000 patients enrolled (with written informed consent), and we are on target to complete recruitment before the end of 2014. The biorepository holds serum, plasma and DNA from all patients. Liver biopsy tissue surplus to diagnostic requirements is also available. In a subset of patients, PBMCs have been isolated and stored. Access to Data and Samples: Access to data and samples is managed by a Tissue and Data Access Committee who have the authority to grant ethical approval for research using the resource. Application for access to samples and data can be made on-line at www.hcvresearchuk.org. We invite applications to use this unique and valuable resource from all interested parties. Funding for the resource has been provided by the UK Medical Research Foundation.

Disclosures:

Graham R. Foster - Advisory Committees or Review Panels: GlaxoSmithKline, Novartis, Boehringer Ingelheim, Tibotec, Chughai, Gilead, Janssen, Idenix, GlaxoSmithKline, Novartis, Roche, Tibotec, Chughai, Gilead, Merck, Janssen, Idenix, BMS; Board Membership: Boehringer Ingelheim; Grant/Research Support: Chughai, Roche, Chughai; Speaking and Teaching: Roche, Gilead, Tibotec, Merck, BMS, Boehringer Ingelheim, Gilead, Janssen

Sharon Hutchinson - Speaking and Teaching: Janssen, Gilead, MSD, Roche

The following people have nothing to disclose: John McLauchlan, Will Irving, John F. Dillon

1447

HCV specific IgG+ memory B cells show strong clonal expansion after spontaneous clearance of an infection with HCV in a single source outbreak

Anne Olbrich1, Hedda Wardemann2, Thomas Berg1, Julia Benck-ert1;

1 University Hostpital Leipzig, Leipzig, Germany;2Max Planck Institute for Infection Biology, Berlin, Germany

In most cases an infection with the Hepatitis C virus (HCV) leads to the development of chronic hepatitis C, whereas spontaneous viral clearance occurs in a smaller percentage of acutely infected individuals. For the successful control of the viral replication in the early phase of infection the adaptive immune system, in particular the humoral response is essential. Several studies have shown that protective anti-HCV antibodies are able to prevent an infection with HCV; however, it is unknown whether differences in antibody repertoire mediate the ability to spontaneously overcome the infection. To characterize differences in antibody response to HCV and to describe potentially HCV neutralizing antibodies we isolated HCV-specific IgG+ memory B cells from peripheral blood of 3 patients with chronic hepatitis C and 3 patients with spontaneous viral clearance of a unique cohort that were infected in a single-source outbreak by an identical HCV-strain. The B cell receptor on the cell surface, representing the antigen specificity of each single B cell, was used to isolate single HCV-specific IgG+ memory B cells by Fluorescence Assisted Cell Sorting (FACS). A RT-PCR based approach was applied to monoclonally express antibodies from these single memory B cells in vitro. Sequence analysis of the amplified individual immunoglobulin chains showed higher numbers of somatic hypermutations as sign of affinity maturation as well as a biased VH repertoire in patients with spontaneous viral clearance. In 288 analysed antibodies of the spontaneous resolvers, we identified 9,7% of clonally expanded sequences with at least a pair of antibodies showing the identical V(D)J rearrangement in heavy and light chain and shared somatic hypermutations. Expression and reactivity testing of clonally expanded antibodies will provide insights to preferentially covered antibody binding sights and may as well offer new therapeutic options as infection prophylaxis by passive immunisation.

Disclosures:

Thomas Berg - Advisory Committees or Review Panels: Gilead, BMS, Roche, Tibotec, Vertex, Jannsen, Novartis, Abbott, Merck; Consulting: Gilead, BMS, Roche, Tibotec; Vertex, Janssen; Grant/Research Support: Gilead, BMS, Roche, Tibotec; Vertex, Jannssen, Schering Plough, Boehringer Ingelheim, Novartis; Speaking and Teaching: Gilead, BMS, Roche, Tibotec; Vertex, Janssen, Schering Plough, Novartis, Merck, Bayer

The following people have nothing to disclose: Anne Olbrich, Hedda Wardemann, Julia Benckert

1448

Exome capture and allinone sequencing to identify potential single nucleotide polymorphisms associated with spontaneous hepatitis C virus clearance

Huiying Rao1, Guangqing Sun2, Dong Jiang1, Fusheng He2, Jianghua Wang1, Yu Xu2, Heng-Hui Zhang1, Xuemei Li2, Ruifeng Yang1, Yinghao Wang2, Haiying Zhang1, Jian Wang2, Degui Sun3, Yingrui Li2, Ruyu Liu1, Jun Wang2, Lai Wei1;

1 Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Peking University People's Hospital, Peking University Hepatology Institute, Beijing, China; 2BGI-Shenzhen, Shenzhen, China; 3Guan County Center for Disease Control and Prevention, Guan, China

Correspondence to Lai Wei, weilai@pkuph.edu.cn; Jun Wang, wangj@genomics.cn Background and Aim: Host IL28B genetic variants have been found to be associated with spontaneous

1449

Changes in characteristics of hepatitis C (HCV) patients referred to liver centers in the United States (US) in the past decade

Nizar Talaat, Suna Yapali, Robert J. Fontana, Hari S. Conjeevaram, Frederick K. Askari, Anna S. Lok

Department of Internal Medicine, Division of Gastroenterology and Hepatology, University of Michigan, Ann Arbor, MI

Background/Aims: The peak of the HCV epidemic in the US occurred in the 1970s. Approval of 2 direct-acting antivirals (DAAs) in 2011 and anticipation of interferon (IFN)-free regimens have led more patients to seek treatment. We compared the characteristics of HCV patients newly referred to our liver clinics in 2011-2 (around the time of approval of the first DAA) with those seen in 1998-9 (just before the approval of pegylated IFN). Methods: Retrospective chart review of clinical and laboratory data was conducted on all HCV patients newly referred to our liver clinics in Era I (1998-9) and Era II (2011-2). Patients with HBV or HIV coinfection or had liver transplantation were excluded. Advanced disease was defined as cirrhosis (based on histology or APRI>2), hepatic decompensation or hCc. Results: A total of 1348 patients (538 in Era I and 810 in Era II) were included. Compared to Era I, patients in Era II were older, more likely to be Black, and had a longer interval between diagnosis and referral. GT1 predominated in both Eras; however, among the patients with GT1, GT1a was more common in Era II (74% vs .62%, P=0.002). A higher percent of patients in Era II were treatment experienced, but 77% had not received any treatment. Patients in Era II were more likely to have advanced disease at referral (62% vs.49%, p<0.001), with an 8-fold higher prevalence of HCC. Despite a longer interval between diagnosis and referral, percent of patients with advanced disease in Era II who had not received HCV treatment was similar to that in Era I (74% vs.81%, p=0.058). Comparison of patients in the two Eras stratified by age found that HCC prevalence in Era II was significantly higher in both patients <50 years (20.1% vs .2%) and those ≥50 years (22% vs.4.4%). Percent of patients who had received treatment in Era II increased in patients <50 years but not those ≥50 years. Conclusion: HCV patients newly referred to liver clinics in the US in recent years are older, more likely to be infected with GT1a, have more advanced liver disease and an 8-fold higher prevalence of HCC compared to a decade ago. Despite being diagnosed for a longer period of time and having access to care, three-quarters of these patients had not received any treatment. Reduction in HCV disease burden will require development of new HCV treatment targeted towards patients in the current Era as well as improvement in access to treatment and experienced providers.

Era I (1998-9) N=538Era II (2011-2) N=810P-value
  1. *Mean±SD

Men, % Age, years* ≥50 years, % Race, % White Black Other Years from diagnosis* Genotype 1 2 3 other Treatment status, % Advanced liver disease, % Compensated cirrhosis,% Decompensated cirrhosis, % HCC%64.9% 46±8 25.1% 85.5% 11.6% 2.9% 3.3±3.7 76.8% 9.6% 11.6% 2.0% 17.4% 49% 28.4% 17.7% 2.6%63.2% 54±11 77.3% 77.6% 17.1% 5.2% 7.4±7.3 79.6% 9.3% 8.7% 2 .4% 23.1% 62% 24.0% 16.4% 21.6%0.53 <0.001 <0.001 0.003 <0.001 0.534 0.013 <0.001 0.065 0.553 <0.001

Disclosure:

Robert J. Fontana - Consulting: GlaxoSmithKline, tibotec; Grant/Research Support: Gilead, vertex, Ocera

Anna S. Lok - Advisory Committees or Review Panels: Gilead, Immune Targeting System, MedImmune, Arrowhead, Bayer, GSK, Janssen, Novartis; Grant/Research Support: Abbott, BMS, Gilead, Merck, Roche

The following people have nothing to disclose: Nizar Talaat, Suna Yapali, Hari S. Conjeevaram, Frederick K. Askari

1450

Evaluation of the Anti-HCV Core High Sensitivity® ELISA Test among Anti-HCV Screening-negative Dialysis Patients at Risk of Occult HCV Infection

Juan A. Quiroga1, Guillermina Barril2 , 1, Dolores Arenas3, Mario Espinosa4, Nuria Garcίa Fernández5, Secundino Cigarran6, Jose Herrero7, Gloria del Peso8, Pilar Caro9, Rebeco Garcίa10, Yesico Amezquita11, Ana Blanco12' Pilar Martlnez13, José M. Alcázor14, Emilio González-Parra15, José C. Dίaz-Bailón16, Adoroción Martίn17, Inmaculada Castillo1, Javier Bartolomé 1, Vicente Correño1

1Fundoción Estudio Hepatitis Virales, Madrid, Spain; 2Nephrology H. U. La Princesa, Madrid, Spain; 3Nephrology, H. Perpetuo Socorro, Alicante, Spain; 4Nephrology, H. U. Reina Sofίa, Córdobo, Spain; 5Nephrology, Clinica Universitaria de Navarra, Pamplona, Spain; 6Nephrology, H. da Costa Burela, Burela, Spain; 7Nephrology, Hospital Clίnico San Carlos, Madrid, Spain; 8Nephrology, H. U. La Paz, Madrid, Spain; 9Nephrology Clίnica Ruber, Madrid, Spain; 10Nephrology, Complejo Hospitalario La Mancha-Centro' Alcázor de San Juan, Spain; 11Nephrology, Clίnico Fuensanta, Madrid, Spain; 12Dial Centro, Madrid, Spain; 13ICN El Pilar, Madrid, Spain; 14Nephrology, Hospital 12 de Octubre, Madrid, Spain; 15Nephrology, Fundoción Jiménez Dίaz, Madrid, Spain; 16FRIAT C. Los Olmos, Segovia, Spain; 17Nephrology, Hospital de Poniente, El Ejido, Spain

Background: Occult HCV infection was reported in anti-HCVand serum HCV RNA-negative hemodialysis (HD) patients but with HCV RNA detectable in PBMC (JASN 2008; 19: 2288-92). More sensitive anti-HCV assays are needed in such special populations. Aim: To evaluate reactivity to anti-HCV Core High Sensitivity® ELISA among dialysis patients at risk of occult HCV infection with abnormal liver enzymes. Patients and Methods: 210 chronic kidney disease patients undergoing dialysis (HD or peritoneal) with abnormal liver enzymes were studied, who resulted repeatedly negative to serum HCV RNA and anti-HCV. All samples were re-tested using one licensed screening assay (Innotest HCV AblV, Innogenetics) and remained anti-HCV-negative. HCV RNA was tested in PBMC and in ultracentrifuged serum (2 ml) by real-time PCR. The anti-HCV Core High Sensitivity® ELISA (DIATER Labs.) is a sensitive assay that detects antibodies to a conserved HCV core epitope region in sera of HCV-exposed persons. According to the supplier, a sample is defined as reactive if the absorbance value is equal to or higher than 1.2 times the cut-off value (referred to as absorbance index, AI). Results: HCV RNA was detectable in PBMC from 42/210 (20%) and in ultracentrifuged serum in 16 (7.6%) other patients; 7 (3.3%) patients resulted positive simultaneously to HCV RNA in PBMC and ultracentrifuged serum. Taken together, 65/210 (30.9%) were classified as having an occult HCV infection (HCV RNA detectable in PBMC and/or ultracentrifuged serum). Anti-HCV Core antibodies were detectable in 11/210 (5.2%) patients, including 4/65 (6.2%) with and 7/145 (4.8%) without HCV RNA detectable in PbMc and/or ultracentrifuged serum. In addition, 44 samples (taken after 1224 months) from 29 patients remaining anti-HCV-screening-negative were tested, including 1 patient with and 28 without baseline anti-HCV Core detectable: 1(3.6%) anti-HCV Corenegative patient at baseline seroconverted to anti-HCV Core. Thus, the anti-HCV Core High Sensitivity® ELISA showed increased antibody reactivity compared with the licensed screening assay (null detection). When the cut-off was lowered and set at Al=1.0, anti-HCV Core reactivity increased up to 8.6% (18/210) including 6/65 (9.2%) patients with virological markers of occult HCV infection. Conclusions: The anti-HCV Core High Sensitivity® ELISA shows an enhanced sensitivity among dialysis patients at risk of occult HCV infection compared with commercial anti-HCV screening assays. Anti-HCV Core testing shows diagnostic usefulness in the management of the dialysis setting because identifies potentially infectious cases without serological or virological markers of HCV infection.

Disclosures:

The following people have nothing to disclose: Juan A. Quiroga, Guillermina Barril, Dolores Arenas, Mario Espinosa, Nuria Garcia Fernandez, Secundino Cigarran, Jose Herrero, Gloria del Peso, Pilar Caro, Rebeca Garcia, Yesica Amezquita, Ana Blanco, Pilar Martinez, Jose M. Alcazar, Emilio González-Parra, Jose C. Dίaz-Bailón, Adoración Martin, Inmaculada Castillo, Javier Bartolomé, Vicente Carreno

1451

Association of tyrosine with insulin resistance in hepatitis C virus-related chronic liver disease

Takahiro Yamasaki, Takashi Oono, Junichi Zaitsu, Issei Saeki, Yoshio Marumoto, Isao Hidaka, Yohei Urata, Tsuyoshi Ishikawa, Taro Takami, Koichi Uchida, Shuji Terai, Isao Sakaida

Department of Gastroenterology and Hepatology, Yamaguchi University Graduate School of Medicine, Ube, Japan

Objective: Insulin resistance (IR) increases during the early stages of hepatitis C virus (HCV)-related chronic liver disease and is a sign of poor prognosis as well as a risk factor for hepatic fibrosis and hepatocellular carcinoma. In liver cirrhosis (LC) patients, the levels of branched-chain amino acids (BCAAs) decrease, whereas levels of aromatic amino acids such as tyrosine (Tyr) and phenylalanine increase. In addition, serum Tyr level has been founded to predict occurrence of diabetes mellitus. However, no clinical studies have examined the relationship between serum Tyr levels and IR in HCV-related chronic liver disease. We aimed to determine the factors affecting IR in HCVrelated chronic liver disease. Patients and Method: We retrospectively examined 71 patients with HCV-related chronic liver disease (chronic hepatitis, 31; LC, 40) and analyzed various parameters, including amino acids, as possible predictors of IR. IR was assessed using the homeostatic model assessment of IR (HOMA-IR). Amino acids were assayed as BCAAs, Tyr level, and the ratio of BCAAs to Tyr level (BTR). Results: There was a significant correlation between HOMA-IR and body mass index (r = 0.40); platelet count (r = -0.29); BTR (r = -0.46, P = 0.0001); prothrombin time (r = -0.36); and levels of hemoglobin (r = -0.26), total bilirubin (r = 0.38), total protein (r = 0.25), albumin (r = -0.53), total cholesterol (r = -0.32), fasting glucose (r = 0.35), and Tyr (r = 0.55, P < 0.0001). However, BCAAs were not significantly correlated with HOMA-IR (r =0.21, P = 0.082). In multivariate analysis, total cholesterol (odds ratio [OR], 6.511; [95% confidence interval (95% Cl), 1.554-27.284; P = 0.010]) and Tyr level (OR, 4.839; 95% Cl, 1.087-21.549; P = 0. 039) were identified as independent parameters contributing to a HOMA-IR of >2.5. Conclusions: Serum Tyr level may be a biomarker of IR in patients with HCVrelated chronic liver disease. It may be important to regulate serum Tyr to control IR in patients with HCV-related chronic liver disease.

Disclosures:

The following people have nothing to disclose: īakahiro Yamasaki, īakashi Oono, Junichi Zaitsu, Issei Saeki, Yoshio Marumoto, Isao Hidaka, Yohei Urata, Tsuyoshi Ishikawa, Taro Takami, Koichi Uchida, Shuji īerai, Isao Sakaida

1452

The Hepatic Expression of Vitamin D Receptor is Inversely Associated with the Severity of Liver Damage in Genotype 1 Chronic Hepatitis C Patients

Salvatore Petta1, Fabio S. Macaluso1, Calogero Cammà 1, Vito Di Marco1, Daniela Cabibi2, Stefania Grimaudo1, Maria Giovanna Minissale1, Rosaria Maria Pipitone1,Antonio Craxi1

1Cattedra ed U. O. C. di Gastroenterologia ed Epatologia, Palermo, Italy; 2Cattedra di Anatomia Patologica, University of Palermo, Italy, Palermo, Italy

Background and aims: Lower 25-Hydroxyvitamin D (25[OH]D) serum levels have been associated with the severity of liver fibrosis in genotype 1 chronic hepatitis C patients (G1CHC), and experimental evidences suggested a liver protective role of vitamin D via interaction with hepatic vitamin D receptor (VDR). We aimed to assess liver VDR protein expression and its association with the severity of liver damage. Methods: Ninety patients with biopsy-proven G1CHC (Scheuer score) and with available frozen liver tissue were consecutively evaluated. Liver VDR protein expression was assessed by western blot analysis. Results: Liver VDR protein expression by western blot progressively reduced from mild to moderate and further to severe necroinflamamntory activity (p<0.001), and from absent-mild, to moderate and further to severe liver fibrosis (p<0.001). By multivariate logistic regression analysis, severe necroinflamamtory activity was independently associated with high triglycerides (OR, 1.025; 95% CI, 1.006-1.044, p=0.008), and low liver VDR protein expression (OR, 0.053; 95%CI, 0.010-0.275; p<0.001), while severe fibrosis with older age (OR, 1.074; 95% CI, 1.011-1.140, p=0.02), low VDR liver protein expression (OR, 0.170; 95%CI, 0.038-0.765, p=0.02), moderate severe steatosis (OR, 3.272; 95%CI, 1.003-10.670; p=0.04), and liver necroinflammatory activity (OR, 2.309; 95%CI, 1.004-5.308; p=0.04). Conclusion: In a cohort of G1 CHC patients, the expression of hepatic VDR protein is inversely and independently associated with the severity of both liver fibrosis and inflammation, translating experimental evidences on human liver, and identifying a new potential therapeutic target for the management of liver damage in CHC.

Disclosures:

The following people have nothing to disclose: Salvatore Petta, Fabio S. Macaluso, Calogero Cammà, Vito Di Marco, Daniela Cabibi, Stefania Grimaudo,

Maria Giovanna Minissale, Rosaria Maria Pipitone, Antonio Craxi

1453

Seminal HCV RNA level may mirror dynamics of plasma HCV RNA in HIV-positive men with acute HCV infection

Daniel Bradshaw1, Francois Lamoury1, Beth Catlett2, Tanya L. Applegate1,John Mcallister2, Gregory J. Dore1, Gail Matthews1, Mark Danta3

1 Kirby Institute, University of New South Wales, Sydney, NSW, Australia; 2St Vincents Hospital, Sydney NSW, Australia; 3Faculty of Medicine, University of New South Wales, Sydney, NSW, Australia

Aims We hypothesise that sexual transmission of hepatitis C virus (HCV) in HIV-positive men who have sex with men may be fuelled by a high semen HCV RNA level in acute or recent HCV (AHCV) infection. Methods The M2000 Abbott RT-PCR was optimised for quantification of HCV RNA in semen with lower limit of detection of 60 IU/ml. Men with AHCV (duration ≤12 months) or chronic HCV (CHCV, >12 months) not currently on anti-HCV therapy were prospectively recruited in Sydney. Paired semen and EDTA plasma samples were assayed for HCV RNA. Results were analysed using Chi2, Mann-Whitney U and Kruskal-Wallis tests. Results Of 53 HCV RNA-positive men, median age 50 (IQR 44-54) years, 13 (24%) had AHCV/HIV, 19 (36%) CHCV/HIV coinfection and 21 (40%) CHCV monoinfection. For men with AHCV, median duration of infection was months with 5/13 (38%) < 3 months. Of 32 HIV-positive men, 27 (84%) were on antiretrovirals with undetectable plasma HIV RNA and median CD4 count 626 (IQR 462-783) cells/mm3. HCV genotypes were 1a (30,57%), 1b (4,7.5%), 3a (15,28%) and other (4,7.5%). Overall 23/53 (43%) men had detectable semen HCV. There was no significant difference between the proportion with detectable semen HCV for AHCV (6/13, 46%) versus CHCV (17/40, 43%) (p=0.82) or for HIVpositive (13/32, 41%) versus HIV-negative (10/21, 48%) men (p=0.62). Median plasma HCV RNA was 6.1 (IQR 5.5-6.5) log IU/ml with no significant difference between the three groups. When detected, median semen HCV RNA was 1.9 (IQR 1.5-2.8) log IU/ml. There was no correlation between magnitudes of HCV RNA in semen and plasma (r2=0.001). There was a trend to higher median plasma HCV RNA in men with detectable versus undetectable semen HCV RNA; 6.2 (IQR 5.8-6.6) log IU/ml versus 5.9 (IQR 5.3-6.3) log IU/ml respectively (p=0.08). However, for AHCV/HIV, median plasma HCV RNA was significantly higher for those with detectable versus undetectable semen HCV RNA; 6.2 (IQR 6.0-6.6) log IU/ml versus 4.8 (IQR 4.6-5.9) log IU/ml respectively (p=0.014). Conclusions HCV RNA was detected in 43% of semen samples with median level 4.2 log IU/ml less than plasma. For men with AHCV/HIV-coinfection, detectable HCV RNA in semen was more likely with a higher plasma HCV RNA, implying a possible relationship between viral dynamics in plasma and semen in the acute phase of HCV. If, as previously described, HlV-coinfected individuals in the early acute phase of HCV have a higher plasma HCV RNA and lower HCV clearance, this could lead to increased semen HCV RNA, facilitating sexual transmission.

Disclosures:

Gregory J. Dore - Board Membership: Bristol-Myers Squibb, Roche, Gilead, Merck, Janssen, Abbvie; Grant/Research Support: Janssen, Bristol-Myers Squibb, Vertex, Roche, Gilead, Merck, Abbvie; Speaking and Teaching: Roche, Merck, Janssen

Gail Matthews - Consulting: Viiv; Grant/Research Support: Gilead Sciences; Speaking and Teaching: BMS, MSD

The following people have nothing to disclose: Daniel Bradshaw, Francois Lamoury, Beth Catlett, Tanya L. Applegate, John Mcallister, Mark Danta

1454

Clinical utility of HCV core antigen quantification in patients with chronic hepatitis C

Stephane Chevaliez, Alexandre Soulier, Lila Poiteau, Jean-Michel Pawlotsky

Virology & INSERM Unit U955, Henri Mondor University Hospital, Creteil, France

Background: Hepatitis C virus (HCV) core antigen has been proposed as a surrogate marker of HCV replication. HCV core antigen detection and quantification is based on a chemiluminescent microparticle immunoassay (CMIA). This assay is automated, two thirds less expensive than HCV RNA level measurement by real-time PCR, not prone to sample carryover contamination, and easier to use than current HCV RNA tests to diagnose chronic hepatitis C, monitor antiviral therapy and be used for broad-scale screening. Objective: We assessed the clinical performance of the Architect HCV Ag assay (Abbott Diagnostics) for the quantification of HCV core antigen in a large number of subjects chronically infected with HCV genotypes 1 to 6. Patients & Methods: 399 patients, including 159 HCV-seronegative and 240 HCV-seropositive patients with quantifiable HCV RNA, were included. The HCV genotype distribution was the following: 62.6%, 6.5%, 12.1%, 17.3%, 0.5% and 1.0% for genotypes 1, 2, 3, 4, 5 and 6, respectively. HCV RNA levels were determined by means of real-time PCR assays including m2000 (Abbott Diagnostics) and Cobas AmpliPrep/Cobas TaqMan HCV test, version 2.0 (CAP/CTM v2.0, Roche Molecular Systems). The HCV core antigen levels were assessed with the Architect HCV Ag assay (Abbott Diagnostics). The results in this assay are expressed in fmol/L and its dynamic range of quantification is 3.0 - 20, 000 fmol/L. Results: Specificity was 100% (95%CI: 98.1%-100.0%). A positive significant correlation was found between HCV core antigen levels measured with the Architect assay and HCV RNA levels, regardless of the real-time PCR assay used (r = 0.90 and 0.92, p < 0.0001 for m2000 and CAP/CTM v2.0, respectively) and of the HCV genotype. HCV core antigen was detectable only if the HCV RNA level was higher than 3 Log10 IU/mL. Conclusion: The Architect HCV antigen assay, which detects and quantify HCV core antigen, is highly specific and easy to perform. It thus represents a valuable screening tool for active HCV infection. However, due to its lack of sensitivity, this assay cannot be used for response-guided antiviral therapy according to current clinical practice guidelines.

Disclosures:

Jean-Michel Pawlotsky - Consulting: Abbott, Achillion, Boehringer-Ingelheim, Bristol-Myers Squibb, Idenix, Gilead, Janssen, Madaus-Rottapharm, Merck, Novartis, Roche; Grant/Research Support: Gilead; Speaking and Teaching: BoehringerIngelheim, Bristol-Myers Squibb, Gilead, Madaus-Rottapharm, Merck, JanssenCilag, Novartis, Abbott

The following people have nothing to disclose: Stephane Chevaliez, Alexandre Soulier, Lila Poiteau

1455

Clinical outcomes of chronic hepatitis C patients related to baseline liver fibrosis stage - a hospital based linkage study

Yi Huang1 , 2, Bastiaan de Boer3, Leon Adams1 , 2, Gerry C. MacQuillon1 , 2, Max K. Bulsara4, Gary P. Jeffrey1 , 2

1School of Medicine and Pharmacology, University of Western Australia, Perth, WA, Australia; 2Department of Gastroenterology and Hepatology, Sir Charles Gairdner Hospital, Perth, WA, Australia; 3Department of Anatomical Pathology, PathWest, QEII Medical Centre, Perth, WA, Australia; 4lnstitute of Health and Rehabilitation Research, University of Notre Dame, Perth, WA, Australia

Background: Long term follow up studies that describe clinical outcomes of chronic hepatitis C (CHC) patients with none, mild or severe liver fibrosis are required to determine cost benefits of anti-viral therapies. This study evaluated long term adverse clinical outcomes for CHC patients stratified by all Metavir fibrosis stages. Methods: Clinical outcomes were determined using population based data linkage methodology for 984 CHC compensated patients who had a liver biopsy performed from to 2012. This included 833 with ongoing infection and 151 with a sustained virological response (SVR). Results: 198 (20.1%) of patients had F0,458 (46.5%) had F1,145 (14.7%) had F2,98 (10%) had F3 and 85 (8.6%) had f4 fibrosis. During 11,226 person-years of follow-up, 31(3.2%) patients developed hepatocellular carcinoma (HCC), 61 (6.2%) developed liver decompensation and 49 (5.0%) had liver transplantation or liver related death (LRD). In the 833 patients with ongoing CHC there was no significant difference in LRD for those with F0, F1 or F2 fibrosis with an 18 year survival probability >94%. Age adjusted hazard ratio (HR) of LRD for F3 compared to F0-F2 was 4.24(P=0.003), with no significant difference in the first 13 year follow up. The 15 year liver complication (HCC and liver decompensation) free survival for F0, F1 and F2 was 100%, 96% and 94% respectively. Age adjusted HR of liver complication free survival for F3 compared to F0-F2 was 3.22 (P=0.001), with no significant difference during the first seven years of follow up. F4 (cirrhosis) had significantly higher risk of LRD, liver decompensation and HCC development than F3 (p<0.001).151 patients had a SVR after HCV treatment and the mean time between biopsy and treatment was 1.8 years. Mean follow up after the SVR was 12 years. O this group 25 (12.6%) patients had F0,75 (16.4%) had F1, 21 (14.5%) had F2, 18 (18.4%) had F3 and 12 (14.1%) had F4. Compared with the group with ongoing HCV infection there was a significant benefit of SVR in F4 patients with HR of 0.15 (95% Cl, 0.02-1.17) for LRD and HR of 0.19 (95% Cl, 0.050.81) for liver complications. There was no improvement in end points for F0, F1, F2 or F3 patients. Conclusion: CHC patients with ongoing infection and F0, F1 or F2 had few liver complications after 15 years follow up. Those with F3 and F4 had significantly increased HR for LRD and liver complications. However for F3 patients the increase in LRD occurred after 13 years and for liver complications after seven years. After a SVR a significant improvement in LRD and liver complications was found only in F4 patients.

Disclosures:

Gary P. Jeffrey - Advisory Committees or Review Panels: MSD, Novartis

The following people have nothing to disclose: Yi Huang, Bastiaan de Boer, Leon Adams, Gerry C. MacQuillan, Max K. Bulsara

1456

Correlation between the levels of TGF beta 1, Foxp3 lymphocytes, bacterial traslocation indexes and liver fibrosis score in HIV, HCV, HIV and HCV coinfected patients and healthy controls

Paolo Sacchi1,Raffaele Bruno1, Serena M. Cima1, Marta Corbella1, Giuditta Comolli2, Antonella Chiesa2, Fausto Baldanti2, Catherine Klersy3, Stefana Novoti1, Mara Mariconti1, Claudio Boldi4

1 Infectious diseases, Foundation IRCCS Policlinico Son Matteo, Pavia, italy; 2Unit of Molecular Virology, Foundation IRCCS Policlinico Son MAtteo, Pavia, Italy; 3Epidemiology service, Foundation IRCCS Policlinico San MAtteo, Pavia, italy; 4Univerty of Milan, Milan, Italy

Background: Previous studies have pointed out that HIV infection accelerates the progression of chronic C hepatitis. Tregs are a subset of CD4 T cells expressing the forkhead-wingedhelix transcription factor (Foxp3). Alterations in Treg cells or in levels of the transforming growth factor beta 1(TGF-beta1), as well of bacterial traslocation markers might be involved in the accelerated course of liver fibrosis characteristically seen in HIV-HCV coinfected individuals. Methodology. A cross-sectional study was conducted on 80 subjects including HIV-monoinfected (n = 20), HCV monoinfected (n = 20) and HIV-HCV coinfected (n = 20) patients, and healthy controls (n = 20) older than 18 years. Foxp3 and TGF-beta 1 levels were measured in peripheral blood and were correlated to liver fibrosis, measured either by biochemical score (FIB 4) or by elastometry, and. We also analysed the correlation of Foxp3 and TGF-beta 1 levels with CD14 (soluble and surface), IL17 and bacterial DNA products (expression of bacterial translocation) Main Findings: Foxp3 % levels were significant higher in HIV+ and HIV-HCV coinfected subjects than in HCV+ and control group (p< 0.0005, Kruskal-Wallis test) There was no statistically significant difference for TGF-beta1 in the four group. FIB 4 values inversely correlated with TGF-beta1 (Rho correlation coefficient −0.38; p=0.0155). as well as with liver stiffness values (Rho correlation coefficient −0.31; p=0.0498. CD14 (soluble and surface) levels were significantly different between HIV+ vs the healthy controls, HIV+HCV+ vs the healthy controls, HCV+ vs HIV+ HCV+ (p< 0.0001, Kruskal-Wallis test). IL17 was significantly different between HCV+ vs the others 3 groups. Bacterial DNA was significantly different in HIV+ vs the others 3 groups Conclusions: Foxp3+ levels are higher in patients with HIV, but they do not influence liver fibrosis staging. TGF-b1 levels inversely correlate with fibrosis suggesting a protective effect. Our data show that the group of HIV- HCV+ has increased levels of bacterial DNA, CD14 (soluble and surface) and IL17 expression of a major translocation as compared with the others groups. The existent correlation between the translocation index and FIB4 suggest that fibrosis stage may depend on immunoactivation caused by bacterial translocation

Disclosures:

The followinq people have nothinq to disclose: Paolo Sacchi, Raffaele Bruno, Serena M. Cima, Marta Corbella, Giuditta Comolli, Antonella Chiesa, Fausto Baldanti, Catherine Klersy, Stefano Novati, Mara Mariconti, Claudio Baldi

1457

Significance of vitamin D receptor gene polymorphisms for risk of hepatocellular carcinoma in chronic hepatitis C

Chao-Hung Hung, Tsung-Hui Hu, Sheng-Nan Lu, Jlng-Houng Wong, Chien-Hung Chen, Chuan-Mo Lee

Division of Hepatogastroenterology, Department of Internal Medicine, Kaohsiung Chrang Gung Memorial Hospital, Kaohsiung, Taiwan

Background/Aims: Biological and epidemiological data suggest that vitamin D levels may influence cancer development. Several single nucleotide polymorphisms have been described in the vitamin D receptor (VDR) gene in association with cancer risk. We aimed to investigate the association of VDR polymorphisms with hepatocellular carcinoma (HCC) development in chronic hepatitis C patients. Methods: In a cross-sectional, hospital-based setting, 340 patients (201 chronic hepatitis, 47 cirrhosis and 92 HCC) and 100 healthy controls receiving VDR genotyping (bat-haplotype: Bsml rs1544410 C, Apal rs7975232 A and Taql rs731236 A) and interleukin (IL)−28B genotyping were enrolled. Results: Patients with HCC had a higher frequency of Apal CC genotype (P=0.018) and bAt[CCA]-hapiotype (P=0.019) as compared to control subjects. There were no differences in Bsml, Taql and IL28B polymorphisms between two groups. In patients with chronic hepatitis C, HCC subjects had a higher frequency of Apal CC genotype and bAt[CCA]-haplotype than those with chronic hepatitis (P=0.001 and 0.002, respectively) and cirrhosis (P=0.019 and 0.026, respectively). After adjusting age and sex, logistic regression analysis showed that Apal CC genotype (odds ratio: 3.02, 95% confident interval: 1.65-5.51) was independently associated with HCC development. Conclusion: VDR Apal polymorphism may play a role in the development of HCC among chronic hepatitis C patients. Further explorations of this finding and its implications are required.

Disclosures:

The followinq people have nothinq to disclose: Chao-Hung Hung, Tsung-Hui Hu, Sheng-Nan Lu, Jlng-Houng Wang, Chien-Hunq Chen, Chuan-Mo Lee

1458

High Spontaneous Clearance of Symptomatic Iatrogenic Acute Hepatitis C Genotype 4 Infection

Mohamed A. Hashem2 , 3, Hassan E. Zaghla1,Zainab Zakaria3, Walaa Ramadan3 , 4, Nabiel N. Mikhail3, Maha Sobhy3, Gehan G. Galal3, Iman Galal3, Sayed F. Abdelwahab3 , 5, Imam Waked1

1HepatoIogy, National Liver Institute, Menoufiya, Egypt; 2Epidemiology and Public Health, University of Maryland School of Medicine, Baltimore, MD; 3Egyptian Company for Blood Transfusion Services (EgyBlood), VACSERA, Giza, Egypt; 4South Egypt Cancer Institute, Assiut University, Assiut, Egypt; 5MicrobioIogy and Immunology, Faculty of Medicine, Minia University, Minia, Egypt

Background: Acute hepatitis C (AHC) resolves spontaneously in 15-40% of patients. Predicting spontaneous resolution is important to identify patients who will need antiviral therapy. Predictors of cure & spontaneous resolution in AHC caused by infection with HCV genotype 4 have not been prospectively studied. This study investigated spontaneous viral clearance in patients with iatrogenically acquired symptomatic AHC. Methods: 26 Patients with symptomatic AHC who had acquired the infection through an identified previous medical procedure were enrolled in a longitudinal observational study since June 2011. AHC was diagnosed in patients with symptomatic acute hepatitis (elevated transaminases >10 times upper limit of normal, with or without jaundice) being HCV-RNA positive, having no antibody to HCV on initial evaluation & developing antiHCV antibody during follow-up. All other causes of acute hepatitis were excluded. Patients were followed weekly in the first month & monthly for 5 further months, with a follow-up visit 6 months after the last RNA negative sample. IL28B SNPs at rs12979860 & HCV-genotype were tested at baseline, & HCVRNA was tested by RT-PCR during each visit. Patients who remained RNA positive at 24 weeks were treated with pegylated interferon & ribavirin for 24 weeks. Results: 17 Patients with iatrogenically acquired AHC (41% following recent surgery, 29% blood transfusion, 24% dental procedure, 5% cystoscopy) completed 6 months follow-up, to either spontaneous resolution or start of treatment. Mean age was 38.0±12 years, 71% were females, with a mean incubation period of 6 weeks (IQR: 2.25-7.00). Viral clearance, with undetectable HCV RNA for at least 24 weeks, occurred spontaneously in 13 (/6%). The average time to resolution was 1/.9±9.3 weeks (range 4-30 weeks). Four patients received therapy, with 2 achieving SVR & are still being treated. The remaining 9 AHC subjects are currently under follow up. No significant differences in rate of spontaneous viral clearance were observed in patients with different IL28B genotypes. All variables tested in the multivariate regression analysis did not achieve levels of significance. Therefore, predicting spontaneous viral clearance after iatrogenic AHC exposure was not possible in this studied population. Conclusion: Patients with symptomatic AHC caused by an iatrogenic exposure had a very high rate of spontaneous resolution in this group. The high spontaneous resolution rate did not allow detecting predictors for spontaneous resolution. This suggests that the clearance rate of symptomatic AHC may be higher than previously reported, especially in iatrogenically acquired genotype 4 infection.

Disclosures:

Imam Waked - Speaking and Teaching: Hoffman L Roche, Merck, Bayer, BMS

The following people have nothing to disclose: Mohamed A. Hashem, Hassan E. Zaqhia, Zainab Zakaria, Walaa Ramadan, Nabiel N. Mikhail, Maha Sobhy, Gehan G. Galal, Iman Galal, Sayed F. Abdelwahab

1459

The use of ALT as screening marker for chronic HCV Liver Disease

Jennifer E. Layden, William H. Adams, Eric R. Kallwitz, Steve Scaglione, Ramon A. Durazo-Arvizu, Guichan Cao, Thomas Layden, Scoff Cofler

Loyola University Medical Center, Maywood, IL

Background: While alanine aminotransferase (ALT) is often used to assess for liver inflammation, there is uncertainty regarding how ALT levels vary by gender, race, and age in persons without liver disease, as well as the utility of ALT as a screening tool. The purpose of this study, therefore, was to use the National Health and Nutrition Examination Survey (NHANES) dataset to identify co-variables that impact average values and determine appropriate cut-off values for screening purposes. Methods: The NHANES dataset was used to define a cohort with no known liver disease (N=3994) using the following exclusion criteria: HCV, HBV, HIV infected; alcohol intake >2 drinks/day for men, >1/day for women; transferrin saturation>45%, elevated serum ferritin, or met any of metabolic syndrome parameters: This cohort was used to estimate the impact of gender, race, and age on average ALT levels and determine stratified marginal means. A chronic HCV cohort was defined as subjects with a positive HCV Ab and detectable HCV viral levels (N=358). Receiver Operative Curve (ROC) analyses were conducted to estimate cut-off values for ALT that best predict chronic HCV infection, both for the whole cohort, and stratified by gender. Results: For the non-liver disease group, females had lower ALT values compared to males across all age groups, and non-hispanic blacks had lower values, especially in women. Marginal means are displayed below, with the non-hispanic white group serving as the reference race group. In AUC analyses, ALT was an excellent marker to identify individuals with HCV in comparison to persons without evidence of liver disease, with AUC's of 0.92 for the overall group, and 0.93 (p<0.001) when stratified by gender. The best ALT cut-off to predict active HCV infection was 21 for women, and 26 for men. Conclusions: Age, race, and particularly gender, impact ALT values in a non-liver disease cohort. ALT is a good screening tool to identify subjects with chronic HCV infection, with cut-off values that are substantially lower that normal reference range values. Such results should be validated for other forms of chronic liver disease.

Marginal Means By Race, Age, and Gender

Age 18-35Age 36-55Age>55
MenWomenMenWomenMenWomen
Mexican American26.6323.9531.7919.6620.8918.99
(1.63)(0.79)(1.93)(1-14)(1.12)(2.27)
Non-Hispanic23.4818.1226.0620.3522.6819.70
White(0.56)(0.33)(0.69)(0.54)(0.51)(0.66)
Non-Hispanic23.2715.5723.9020.6819.4015.95
Black(1.06)(0.34)(0.78)(1.85)(0.86)(1.05)

Disclosures:

Scott Cotler - Speaking and Teaching: Genentech, Vertex, Brystal Myers, Gilead

The following people have nothing to disclose: Jennifer E. Layden, William H. Adams, Eric R. Kallwitz, Steve Scaglione, Ramon A. Durazo-Arvizu, Guichan Cao, Thomas Layden

1460

IL28B gene polymorphysms and US liver fatty changes in patients who spontaneously cleared hepatitis C virus infection

Gloria Taliani1, Martina Spaziante1, Elisa Biliotti1, Marina Borro2, Donatella Palazzo1,Stefania Grieco1,Cristiana Franchi1,Giancarlo laiani1, Caterina Furlan1, Valentina Gallinaro1, Maurizio Simmaco2

1 Clinical Medicine, Sapienza University of Rome, Rome, Ifoly; 2Neuroscienze, Salute Mentale, Organi di Senso, Sapienza University of Rome, Rome, Italy

BACKGROUND: Recent clinical studies have shown that the presence of CC genotype in the rs12979860 region of IL28B gene is associated with an increase in the probability of spontaneous clearance of hepatitis C virus (HCV). Moreover, IL28B polymorphism seems to influence the probability of developing liver steatosis in chronic HCV patients. AIMS: The aims of our clinical study were 1)to verify the distribution of IL28B genotypes (CC, CT or TT) among subjects with spontaneous clearance of HCV infection and 2) to examine the correlation between IL28B polymorphism and hepatic steatosis among these subjects. METHODS AND PATIENTS: We enrolled 41 subjects with spontaneous resolution of HCV infection (detectable serum anti-HCV but undetectable HCV-RNA) and 134 healthy controls from the same geographical area. The IL28B single-nucleotide polymorphism (SNP) rs12979860 was genotyped by using a Pyrosequencing™ technique. The presence of steatosis was assessed by liver biopsy or ultrasound examination in the 41 study subjects. RESULTS: CC, CT and TTgenotypes of the SNP rs1979860 were found in 66%, 24% and 10% of the subjects who spontaneously cleared HCV and in 31%, 54% and 15% of controls, respectively (p=0.0003). Among the study subjects, females with CC-genotype were significantly more represented (p=0.02). Hepatic steatosis did not correlate with IL28B genotype (p=0,14) but only with a high body mass index (BMI) value (p=0.03). CONCLUSIONS: Female subjects carrying IL28B CC-genotype are significantly more represented among Italian patients who spontaneously cleared HCV infection. In addition, among these subjects, the presence of liver steatosis does not correlate with IL28B genotype but is solely related to the occurrence of high BMI. Thus, the association between IL28B polymorphism and steatosis in chronic HCV patients requires the presence of active HCV replication to occur, while in subjects who have cleared the infection, the mechanism(s) inducing liver steatosis are independent from IL28B profile.

Disclosures:

Gloria īaliani - Advisory Committees or Review Panels: Roche, BMS, Gilead, Merck, Janssen; Speaking and Teaching: Roche, BMS, Gilead, Merck, Janssen, Novartis

The following people have nothing to disclose: Martina Spaziante, Elisa Biliotti, Marina Borro, Donatella Palazzo, Stefania Grieco, Cristiana Franchi, Giancarlo Iaiani, Caterina Furlan, Valentina Gallinaro, Maurizio Simmaco

1461

Prediction of Disease Severity in Chronic Hepatitis C: Clinical Gestalt And Role of Transient Elastography

Naveen Gara4,1, Elizabeth C. Wright2, Nalini K. Sharma4'3, Christopher Koh4, David E. Kleiner5, Averell H. Sherker6, Jay H. Hoofnagle6, Marc G. Ghony4

1Georgetown University/ Washington Hospital Center, Washington, DC; 2Office of the Director, NIDDK, NIH, Bethesda, MD; 3Department of Gastroenterology, Kaiser Permanente, Bethesda, MD; 4Liver Diseases Branch, NIH, Bethesda, MD; 5Laboratory of Pathology, NIH, Bethesda, MD; 6Liver Diseases Research Branch, NIH, Bethesda, MD

BACKGROUND: Liver biopsy is often recommended to aid in the decision whether to recommend therapy in patients with chronic hepatitis C (CHC). We prospectively evaluated the accuracy of clinical assessment of liver fibrosis and cirrhosis with and without results of a non-invasive test of liver fibrosisultrasound transient elastography (TE). AIMS: To assess the accuracy of clinical gestalt for prediction of cirrhosis and the utility of TE in improving accuracy in patients with CHC. METHODS: Over an 18 month period, consecutive, consenting adult patients with CHC who were scheduled to undergo liver biopsy were recruited. Each subject was interviewed and examined independently by a junior and a senior hepatologist. Examiners had access to routine pre-biopsy lab tests and made an assessment of fibrosis severity as either: 0 (mild, Ishak 0-2), 1(moderate, Ishak 3-4) or 2 (severe, Ishak 5-6). TE was then performed with the examiners being blinded to each other's results. Based upon the TE results, the initial estimate of liver fibrosis could then be revised. At the end of the study, all clinical and lab data were shown to an expert hepatologist in a standardized format for assessment of liver fibrosis as 0, 1 or 2. After the initial assessment, the reviewer was given TE scores from both examiners to re-assess disease stage. Liver biopsies were scored by the same hepatopathologist using the Ishak staging system. Examiners were blinded to biopsy results until completion of the study. Weighted-Cohen's kappa was used as a measure of agreement between estimated and biopsy determined stage. RESULTS: 98 patients were enrolled in to the study. Mean age was 54 years, 56% were male, 56% were Caucasian and 27% African-American. Mean BMI was 27 and 72% were genotype 1.84 patients were included in the final analysis; 14 were excluded due to cancelled biopsy (n=6), failed TE exam (n=7) or both (n=1). By histologic stage 67% were Ishak 0-2, 20% Ishak 3-4 and 13% Ishak 5-6. On initial clinical assessment, the kappa coefficient between the junior hepatologist and biopsy stage was 0.48 which improved to 0.62 after TE. Results for the senior hepatologist were 0.61 before and 0.58 after TE. The initial kappa for the reviewing hepatologist was 0.53, which improved to 0.63 after TE. Diagnosis of cirrhosis was correct by clinical assessment 73-82% of cases and 91-100% after TE. TE correctly identified all cases of cirrhosis. Inter-operator correlation for TE was 0.85. CONCLUSION: Clinical assessment of cirrhosis was excellent but varied by the level of experience. TE is a useful adjunct for diagnosis of cirrhosis and less-experienced clinicians benefitted more from its use.

Disclosures:

The followinq people have nothinq to disclose: Naveen Gara, Elizabeth C. Wright, Nalini K. Sharma, Christopher Koh, David E. Kleiner, Averell H. Sherker, Jay H. Hoofnagle, Marc G. Ghany

1462

Hepatitis C Screening Rates at a Single Center after the Release of a CDC Recommendation to Screen All Adults Born Between 1945 and 1965

Chris Albers, Amir A. Qamar, Maureen A. Tellier, Fredric D. Gordon

Transplant & Hepatobiliary Diseases,Lohey Hospital and Medical Center, Burlington, MA

Purpose: There are an estimated 1.2 million Americans born between 1945 and 1965 infected with hepatitis C (HCV), but are unaware of their disease. In August 2012 the CDC recommended one-time testing for HCV in this cohort. Methods: HCV antibody (Ab) or PCR testing was evaluated in all patients born from 1945 to1965 seen in general medicine clinics in our tertiary healthcare system each month beginning April 2012. Data from August of 2012 was excluded. Patients with a prior HCV diagnosis were excluded. Patients were categorized into 2 groups: “Known/negative" was defined as a negative HCV test >14 days prior to the clinic visit (test data was available beginning January 1, 2005); “Unknown/assessed" was defined as no previous HCV testing, but testing was performed between 14 days prior to the visit and 30 days after the visit; “Unknown/not assessed" was defined as no previous HCV testing and no HCV testing performed <30 days following the visit. Results: In the 4 months preceding the CDC recommendation a mean of 6, 1/3 unique patient visits occurred each month.13.8% of the patients were known/negative.1.1% of the patients were unknown/assessed (see figure).4 patients were found to be HCV Ab positive but only 1 was PCR positive. In the months following the recommendation a mean of 7,444 unique patient visits occurred per month. The percentage of patients known/negative increased to 16.3% in the last month of data. Within 2 months of the recommendation the percentage of patients unknown/assessed peaked at 2.6% and subsequently decreased to 1.7% in the last month of data.9 patients were found to be HCV Ab positive and none were PCR positive. Conclusions: The release of the CDC recommendation has had little impact on HCV screening in primary care clinics. HCV status is unknown in more than 80% of patients in this cohort seen each day yet only between 1 and 2% of these patients are then screened for HCV.

image

Disclosures:

Fredric D. Gordon - Advisory Committees or Review Panels: Vertex, Gilead; Grant/Research Support: Vertex, Gilead; Speaking and Teaching: Merck

The following people have nothing to disclose: Chris Albers, Amir A. Qamar, Maureen A. Tellier

1463

The MICA but not the DEPDC5 polymorphism is associated with chronic hepatitis C-related hepatocellular carcinoma

Hoang Hai, Akihiro Tamori, Kanako Yoshida, Atsushi Hagihara, Etsushi Kawamura, Hideki Fujii, Sawako K. Uchida, Shuji Iwai, Hiroyasu Morikawa, Masaru Enomoto, Yoshiki Murakami, Thuy T. Le, Norifumi Kawada

Departments of Hepatology, Osaka City University Graduate School of Medicine, Osaka, Japan

Background: Chronic infection with hepatitis C virus (HCV) is closely related to hepatic fibrosis and hepatocellular carcinoma (HCC), but the clinical course of HCC development differs among patients. Recently, DEPDC5 rs1012068 and MICA rs2596542 genetic variations were identified to associate with HCV-related HCC by two independent genome-wide association studies in two different Japanese populations. However, in a Caucasian population, only the MICA single nucleotide polymorphism (SNP) was associated with HCC development. The aim of the present study was to determine whether these SNPs are predictive of HCC development in a unique Japanese population of chronic hepatitis C (CHC) patients. Methods: A total of 800 CHC patients (141 HCC cases and 659 non-HCC controls) from the Osaka area were enrolled in the study from May 2003-March 2013. Genotyping of DEPDC5 rs1012068 and MICA rs2596542 SNPs was performed using a ĪaqMan SNP genotyping and direct sequencing methods. Results: The major, heterozygous, and minor genotypes of the DEPDC5 SNP were found in 42, 93, 6 HCC patients and 173, 474, 12 non-HCC patients, respectively. We did not find a significant difference between DEPDC5 genotype and HCC development (P = 0.1235). This result is consistent with a previous study in a Caucasian population but differs from results in a Japanese population. However, the minor genotype of the MICA SNP was found in 18.44% (26/141) of HCC patients and 11.38% (75/659) of non-HCC patients, and was significantly associated with HCC development (P = 0.022; odds ratio =1.76). Interestingly, the ratio of HCC to non-HCC patients with the minor genotype of MICA rs2596542 was higher than that of non-minor genotypes in all 5-year age groups, and increased proportionally with age. Conclusions: The minor genotype of MICA rs2596542 correlates with an increased risk of HCC development, particularly in older patients.

Disclosures:

Akihiro Tamori - Grant/Research Support: MSD

The following people have nothing to disclose: Hoang Hai, Kanako Yoshida, Atsushi Hagihara, Etsushi Kawamura, Hideki Fujii, Sawako K. Uchida, Shuji Iwai, Hiroyasu Morikawa, Masaru Enomoto, Yoshiki Murakami, Thuy T. Le, Norifumi Kawada

1464

Simple way to use non-invasive tests for cirrhosis diagnosis

Paul Cales1, Jerome Boursier1, Frederic Oberti1, Isabelle FouchardHubert1, Sandrine Bertrais1, Vincent Leroy2

1 Hepatology Department, Centre Hospitalier Universitaire d' Angers, Angers Cedex 9, France; 2Hepatology, CHU, Grenoble, France

Introduction. Several trials, especially in chronic hepatitis C, rely cirrhosis diagnosis on a single cut-off of non-invasive test(s). False positives are generally thought to be fibrosis stage(s) close to cirrhosis. Yet, these statements are based on any recommendation. Therefore, we evaluated predictive values for cirrhosis of available non-invasive tests including a detailed fibrosis classification. Methods. All 1735 patients had chronic hepatitis C and liver biopsy with Metavir fibrosis (F) staging. We evaluated negative (NPV) and positive (PPV) predictive values of tests considering either only the F4 class or all the classes including F4 (e. g. F3/F4) called Fx/4. The highest value of NPV and PPV determined the choice of fibrosis class cut-off and non-invasive test. In population #1 including 1056 patients, we compared blood tests: Fibrotest, FibroMeter and CirrhoMeter. In population #2 including 679 patients, we compared previous blood tests, liver stiffness (Fibroscan) and their combination (CombiMeter). Other characteristics were evaluated: F distribution, morphometry, markers of liver function or portal hypertension. Results (table). Population #1: considering a cirrhosis trial, the optimal choice relies on the cut-off of CirrhoMeter F4 class since its PPV provides a high inclusion rate of cirrhosis (88%) vs. a rate of only 37% with Fibrotest (35% of pts being F2 or F1 or F0), but at the expense of a higher number of patients to screen. Considering trials excluding cirrhosis, the optimal choice relies on the cut-off of CirrhoMeter Fx/4 classes since its NPV provides a low inclusion rate of cirrhosis (1%) vs. a rate of 4% with Fibrotest, but at the expense of a higher number of patients to screen. Population #2: results validated the best PPV of CirrhoMeter F4 class (89%). They also validated an excellent NPV of Fx/4 classes in all single tests (NPV=97%) with, nevertheless, a small advantage for the test combination (NPV: 98%). Conclusion. A blood test designed for cirrhosis can affirm (88-89% prediction) or exclude (97-99% prediction) cirrhosis by using different cut-offs of a detailed fibrosis classification. This can be easily applied in trials whereas, in clinical practice, another examination might be required in the grey zone between the two cut-offs. Certain criteria induce the inappropriate inclusion of around 2/3 of patients.

Population: F classesFibrotestFibroMeterCirrhoMeterFibroscanCombiMeter
PPVNPVPPVNPVPPVNPVPPVNPVPPVNPV
#1: F437.195.379.290.488.092.6----
#l: Fx/435.195.633.098.032.598.8----
#2: F434.394.466.786.888.989.078.889.288.289.7
#2: Fx/432.894.531.096.831.596.747.196.946.898.2

Disclosures:

Paul Cales - Consulting: BioLiveScale

Frederic Oberti - Speaking and Teaching: LFB, gore

Isabelle Fouchard-Hubert - Speaking and Teaching: JANSSEN

Vincent Leroy - Board Membership: roche, merck, gilead, bms, roche, merck, gilead, bms, roche, merck, gilead, bms, roche, merck, gilead, bms; Consulting: jansen, jansen, jansen, jansen; Grant/Research Support: roche, gilead, bms, roche, gilead, bms, roche, gilead, bms, roche, gilead, bms; Speaking and Teaching: bms, merck, gilead, roche, bms, merck, gilead, roche, bms, merck, gilead, roche, bms, merck, gilead, roche

The following people have nothing to disclose: Jerome Boursier, Sandrine Bertrais

1465

The detection and correction of unreliable results of combined liver fibrosis tests increases accuracy up to 93%

Paul Cales1, Gilles Hunault1, Jerome Boursier1, Frederic Oberti1, Isobelle Fouchard-Hubert1,Jean-Pierre H. Zarski2

1Hepatology Department, Centre Hospitalier Universitaire d' Angers, Angers Cedex 9, France; 2Hepatology, CHU, Grenoble, France

Introduction. Liver fibrosis tests are becoming popular. They have still some limits: residual misclassification rate and indication errors in clinical practice. Our aim was to make accuracy higher and more robust in clinical practice by evaluating reliability and correcting unreliable results. Methods.729 patients with chronic hepatitis C were included in a prospective study. They had 6 blood tests, liver stiffness (Fibroscan) and liver biopsy. Metavir fibrosis staging was the reference for accuracy (correct classification rate) of fibrosis tests expressed in multin-omial fibrosis classes. The statistical method included the 3 following steps: 1/ Accuracy improvement: multivariate analysis of available fibrosis markers provided a new test designed for significant fibrosis and combining 8 markers (blood markers and liver stiffness) called CBM.2/ Reliability determination: iterative logistic regressions individualized patient subgroups (or reliable classes) with significantly different CBM accuracy by independent predictors among available CBM markers.3/ Unreliability correction: in CBM subgroups judged unreliable (accuracy <90%), CBM was replaced by the most accurate fibrosis test among those available with the 8 CBM markers. Results. Step 1 provided a CBM test with accuracy at 91.2% (p<0.001 vs each single test). Step 2 provided 8 reliability classes with the following increasing accuracy (prevalence): 0% (0.6%) to 33.3% (0.4%), 50.0% (0.9%), 71.4% (2.1%), 84.8% (13.7%), 90.0% (4.3%), 94.1% (73.0%) and 100% (4.9%), p<O. o01 by ANOVA. Accuracy in the 5 unreliable classes was 77.6% vs 94.2% in the 3 reliable classes (p<0.001). Step 3 of CBM replacement increased accuracy from 77.6% to 88.8% (p<0.001 by McNemar test) in the 17.7% of patients with previously unreliable classes. Thus, overall accuracy was increased from 91.2% (raw CBM accuracy) to 93.3% (corrected CBM accuracy) i. e. a 2.1 % gain. Finally, these “academic” results were translated into clinical practice through a simulation. Raw and corrected accuracy were respectively, whether the simulated prevalence of unreliable CBM results was multiplied by 2: 89.3% vs 92.6% (gain: 3.3%) or multiplied by 3: 84.9% vs 91.2% (gain: 6.3%). Conclusion. Thanks to reliability assessment and correction of unreliable results, the present combination of blood markers and elastometry improves accuracy, and guaranties a high accuracy in clinical practice conditions exposed to numerous unreliability causes, especially comorbidities.

Disclosures:

Paul Cales - Consulting: BioLiveScale

Frederic Oberti - Speaking and Teaching: LFB, gore

Isabelle Fouchard-Hubert - Speaking and Teaching: JANSSEN

Jean-Pierre H. Zarski - Advisory Committees or Review Panels: BMS, Gilead, Janssen Cilag, BMS, Gilead, Janssen Cilag; Consulting: Roche, Scherring Plough, Novartis, Roche, Scherring Plough, Novartis; Speaking and Teaching: Siemens

The following people have nothing to disclose: Gilles Hunault, Jerome Boursier

1466

Acoustic Radiation Force Impulse is better than Transient Elastography in assessing liver fibrosis in chronic hepatitis C using Collagen Proportionate Area as reference

Vincenza Calvaruso, Vito Di Marco, Fabrizio Bronte, Maria Grazia Bavetta, Francesco Rini, Nicola Alessi, Calogero Cammá, Antonio Craxi

Gastroenterologia & Epatologia, DIBIMIS, University of Palermo, Palermo, Italy

BACKGROUND; Liver stiffness, measured by Transient Elastography (TE) or by Acoustic Radiation Force Impulse ARFI, correlates to the stage of fibrosis at biopsy, but is also affected by necroinflammation. Since Collagen Proportionate Area (CPA) is a continuous histological variable measuring collagen but not necroinflammation, it could represent a better reference to assess the performance of TE and ARFI in the setting of noninvasive staging of fibrosis. METHODS: Ninety-three consecutive patients with chronic hepatitis C (CHC) were evaluated for histological fibrosis (METAVIR score), CPA measurement and biochemical features, and underwent TE and ARFI. RESULTS: TE was unreliable in six patients (6.4%), while ARFI measurement was recorded in all patients. By linear regression analysis both TE and ARFI significantly correlated with CPA (CPA-ARFI: R2 = 0.522 p < 0.001; CPA-TE: R2 = 0.454 p < 0.001). By univariate analysis AST, PLT, TE, ARFI, inflammation grade and CPA were related to MetAvIR stage > 2. At multivariate logistic regression analysis, only CPA (OR: 1.47, CI95%: 1.07-2.01, p=0.01 8), inflammation grade (OR: 5.42, CI95%1.06-27.70, p = 0.042) and ARFI (Oade and CPA were related to cirrhosis (METAVIR stage F4), but by multivariate analysis, only CPA (OR: 1.66, CI95%: 1.23-2.23, p=0.001) and ARFI (OR: 29.87, CI95%: 2.25-397.45, p=0.010) were independently associated with cirrhosis. Liver stiffness by TE was not independently associated with METAVIR stage ≥ F2 (OR: 1.14, CI95%: 0.83-1.57, p=0.416) but was marginally associated with cirrhosis (OR:.2.60, CI95%: 0.86-7.85, p=0.091). CONCLUSIONS: In patients with CHC, liver stiffness evaluations by TE and ARFI are related to CPA. However, ARFI imaging is more accurate than TE for the non-invasive staging of both significant and severe stages of liver fibrosis.

Disclosures:

The following people have nothing to disclose: Vincenza Calvaruso, Vito Di Marco, Fabrizio Bronte, Maria Grazia Bavetta, Francesca Rini, Nicola Alessi, Calogero Camma, Antonio Craxi

1467

Storage conditions of plasma samples do not significantly influence reported HCV RNA results in a low viremic setting

Benjamin Maasoumy1, Gavin A. Cloherty2, Birgit Bremer1, Michael P Manns1, Markus Cornberg1, Heiner Wedemeyer1

1Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany; 2Abbott Molecular, Des Plaines' IL

Background: In chronic hepatitis C virus (HCV) infection ontreatment HCV RNA measurement is one of the most important tools to monitor response to antiviral therapies. With the development of direct acting antivirals (DAAs) and the introduction of response guided therapy patients are eligible for truncated therapy or treatment futility based on detectability of very low levels of HCV RNA at defined time points. As a result, accuracy in assessing HCV viral load at the low end of the dynamic range for commercially available assays can have a tremendous impact on treatment decisions. However, there is a huge variability in plasma collection, storage and shipment to the laboratory. It is unknown to what extend different storage and transport conditions influence HCV RNA test results in samples with low level viremia. Methods: In order to mimic a low viremic setting we created plasma samples with targeted HCV RNA levels of 4-260 IU/ml by using human plasma and the WHO 2nd International HCV RNA standard. Aliquots of these samples were incubated at 4°C, 25°C (room temperature; RT) and 37°C for 2h, 6h, 12h, 24h, 72h, or up to 7d. In each aliquot HCV RNA was measured three times using the Abbott RealTime HCV viral load assay. Results: HCV RNA was stable at 4°C, 25°C and 37°C even after 7 days of incubation. In samples with low but still quantifiable HCV RNA, HCV viral loads measured after 3-7 days of storage at 4°C and 25°C were nearly equal to baseline levels (mean decline: −0.06log and −0.11log, respectively) and showed only a small decrease if incubated at 37°C (−0.2log). For those samples with an HCV RNA level below the limit of quantification or detection, HCV RNA was detected in 73% at baseline and in 67%, 58% and 79% of cases after storage at 4°C for 2-6h, 12-24h and 3-7d, respectively. In contrast, a small difference occurred after incubation at room temperature (58%, 67% and 58%) and at 37°C (50%, 56% and 67%). However, there was either exactly the same (25°C; RT) or even a higher (37°C) number of detected HCV RNA results after long incubation for 3-7d compared with only a short incubation period of 2-6h. Conclusions: Storage of HCV RNA plasma samples at 4°C, RT or 37°C for up to 7 days does not have a significant impact on the reported HCV RNA result. Thus according to our data it seems unlikely that treatment decisions are influenced by plasma storage and transport conditions. We are currently investigating HCV RNA stability in serum and full blood as well as in samples obtained from patients receiving antiviral treatment. Data will be presented at the meeting.

Disclosures:

Benjamin Maasoumy - Advisory Committees or Review Panels: Abbott Molecular; Speaking and Teaching: MSD, Roche Diagnostics, Roche Pharma

Gavin A. Cloherty - Employment: Abbott Molecular; Stock Shareholder: Abbott Laboratories

Michael P. Manns - Consulting: Roche, BMS, Gilead, Boehringer Ingelheim, Novartis, Idenix, Achillion, GSK, Merck/MSD, Janssen, Medgenics; Grant/Research Support: Merck/MSD, Roche, Gilead, Novartis, Boehringer Ingelheim, BMS; Speaking and Teaching: Merck/MSD, Roche, BMS, Gilead, Janssen, GSK, Novartis

Markus Cornberg - Advisory Committees or Review Panels: Merck (MSD Germamny), Roche, Gilead, Novartis; Grant/Research Support: Merck (MSD Germamny), Roche; Speaking and Teaching: Merck (MSD Germamny), Roche, Gilead, BMS, Novartis, Falk

Heiner Wedemeyer - Advisory Committees or Review Panels: Transgene, MSD, Roche, Gilead, Abbott, BMS, Falk; Grant/Research Support: MSD, Novartis, Gilead, Roche, Abbott; Speaking and Teaching: BMS, MSD, Novartis, ITF

The following people have nothing to disclose: Birgit Bremer

1468

Liver Transplantation for Patients with Pretransplant Undetectable Hepatitis C RNA: Can Eradication of Virus Guarantee Superior Outcome?

Shunji Nagai1, Gabriel Schnickel1,loannis Theodoropoulos1, David A. Bruno1, Marwan Kazimi1, Kimberly Ann Brown2, Atsushi Yoshida1,Marwan S. Abouljoud1

1TranspIant and Hepatobiliary Surgery, Henry Ford Hospital, Detroit, MI; 2Gastroenterology and Hepatology, Henry Ford Hospital, Detroit, MI

Background: Eradication of hepatitic C virus (HCV) prior to liver transplantation (LT) in cirrhotic patients is a reasonable strategy to prevent post-transplant HCV recurrence. Undetectable levels of HCV RNA can be found in LT candidates secondary to pharmacologic treatment or spontaneous clearance. An understanding of the post-transplant HCV disease progression and recurrence in this population is still limited. The aim of this study was to investigate the outcomes of LT patients that were HCV antibody positive, but had undetectable HCV RNA at the time of transplant. Methods: A total of 22 patients were found to have undetectable HCV RNA at transplant. Fifteen of these patients received pre-LT anti-viral therapy, of which 9 achieved a sustained virologic response, while spontaneous clearance of HCV was observed in 7 patients. Virologic and histologic recurrence, and overall survival were set as endpoints. Results: Post-LT virologic recurrence occurred in 36% (8 of 22). The rates of histological recurrence (stage 2 or greater fibrosis) at 1-year and 3-years were 29% and 44%, respectively, which was comparable to patients in the pre-transplant detectable HCV RNA group (n =162) (p = 0.31). Advanced fibrosis (stage 3 and 4) occurred in 3 of 22 patients, and no graft loss or mortality secondary to HCV recurrence was observed in the pre-LT undetectable HCV RNA group. Conclusion: The results of this study suggested that, although virologic recurrence was lower, undetectable HCV RNA prior to transplant did not necessarily led to an increase in recurrence free time or patient survival. Comparison of histological recurrence and of HCV and overall survival between the detectable and undetectable HCV RNA before liver transplantation.

(A) Endpoint as stage 2-4 fibrosis recurrence (F2-4) (p = 0.31).

(B)Overall survival (p =0.36).

image

Disclosures:

Kimberly Ann Brown - Advisory Committees or Review Panels: CLDF, Merck, Salix, Gilead, Vertex, Novartis, Genentech, Gilead, Janssen, Novartis, Salix; Consulting: Blue Cross Transplant Centers, Salix; Grant/Research Support: CLDF, Gilead, Exalenz, CDC, BMS, Bayer-Onyx, Ikaria, Hyperion, Merck; Speaking and Teaching: Salix, Merck, Genentech, Gilead, CLDF, Vertex

The following people have nothing to disclose: Shunji Nagai, Gabriel Schnickel, loannis Theodoropoulos, David A. Bruno, Marwan Kazimi, Atsushi Yoshida, Marwan S. Abouljoud

1469

Fluctuations in Serum HCV RNA Levels during the Natural Course of HCV Infection

Hidenori Toyoda, Takashi Kumada, Toshifumi Tada, Takanoni Ito

Gastroenterology, Ogaki Municipal Hospital, Ogaki, Japan

Purpose: Serum hepatitis C virus (HCV) RNA levels are reportedly associated with sustained virologic response to antiviral therapy and are an important predictor of treatment outcome. However, few studies have investigated whether HCV RNA levels fluctuate and whether fluctuations impact the natural course of chronic HCV infection. We investigated the range of HCV RNA fluctuations during the natural course of HCV infection and its clinical impact. Methods: Serum HCV RNA levels were serially measured using real-time PCR methods in 336 patients (138 males and 198 females, 243 genotype 1 infection and 93 genotype 2 infection) every 3 to 6 months for more than 3 years between December 2007 and December 2011. No patients had received antiviral therapy. Fluctuations in HCV RNA levels and its association with clinical features of chronic hepatitis were analyzed. Results: There were a total of 1392 HCV RNA measurements. The median number of measurements per patient was 12 (range, 6-17) with a median interval of 3.4 months (range, 2.6-7.4). Fluctuations during the observation period ranged from 0.3 log10 to 5.4 log10 (median, 0.9 Iog10). The fluctuation was less than 10-fold in 171 patients (50.9%), 10- to 100-fold in 131 patients (39.0%), and 100-fold or more in 34 patients (10.1%). No background factors were associated with greater fluctuation in HCV RNA levels, except for HCV genotype 2 infection, which was associated with a higher percentage of > 100-fold fluctuation (5.8% of genotype and 21.5% of genotype 2, p<0.0001). Average serum alanine aminotransferase (ALT) activity during the study period was higher and average platelet counts were lower in patients with >100-fold fluctuation than those with <100-fold fluctuation (ALĪ, 49.8±29.3 vs.37.1±18.3 lU/mL, p=0.0094; platelets, 135±61 x 103 vs.172±71 x 103, p=0.0031). Multivariate analysis revealed that factors associated with elevated average ALT activity (>35 lU/mL) were HCV genotype 1(odds ratio, 1.71; p=0.0445) and >100-fold fluctuation of HCV (odds ratio, 2.99; p=0.0061), and factors associated with decreased platelet counts (<15 /μL) were higher age (odds ratio, 18.8; p=0.0007) and ≥100-fold fluctuation in HCV (odds ratio, 3.14; p=0.0042). Greater fluctuation was also associated with higher ALT activity and lower platelet counts when comparing patients with and without ≥10-fold fluctuation in HCV. Conclusions: Greater fluctuation in serum HCV RNA levels during the natural course of HCV infection (especially ≥100fold) was associated with higher ALT activity and lower platelet counts and may cause rapid progression of chronic hepatitis C.

Disclosures:

īhe following people have nothing to disclose: Hidenori Toyoda, Takashi Kumada, Toshifumi Tada, Takanori Ito

1470

Gradual increase in platelets following sustained virological response among patients with HCV-induced advanced hepatic fibrosis

Adriaan J. van der Meer1, Raoel Maan1, Bart J. Veldt1, Jordan J. Feld2, Heiner Wedemeyer3, Jean-Francois Dufour4, Frank Lammert5, Andres Duarte-Rojo2, Michael P. Manns3, Stefan Zeuzem6, Wolf P. Hofmann6, Robert J. de Knegt1, Bettina E. Hansen1, Harry L. Janssen1,2

1Gastroenterology & Hepotology, Erasmus MC, Rofferdam, Netherlands; 2Liver Centre, Toronto Western Hospital, Toronto, ON, Canada; 3Deparfmenf of Gostroenterology, Hepafology, and Endocrinology, Medical School Hannover, Hannover, Germony; 4Hepatology, Deparfmenf of Clinical Research, University of Bern, Bern, Switzerland; 5Department of Medicine II, Saarland University Medical Center, Homburg, Germany; 6Medizinische Klinik 1, Klinikum der Johann Wolfgang GoetheUniversim Fronkfurf am Main, Germany

INTRODUCTION Changes in platelets (PLT) have been correlated with changes in hepatic fibrosis among patients with chronic hepatitis C virus (HCV) infection. We assessed the evolution of PLT following sustained virological response (SVR) in patients with HCV-induced advanced fibrosis. METHODS Available PLT counts (x10A9/L) from 24 weeks after the last treatment up to the latest counts prior to cirrhosis-related complications were collected in an international cohort of consecutive patients with chronic HCV infection and advanced hepatic fibrosis (Ishak 4-6) who started interferon-based therapy between 1990 and 2003. Repeated measurement analysis with a random intercept and slope per patient and an unstructured covariance matrix was used to analyze PLT over time, correcting for potential non-linearity. Data are presented as median (interquartile range). RESULTS In total 464 patients were included; 321(69%) patients were male, median age was 51 (44-57) years, and 353 (76%) had cirrhosis. SVR was attained by 187 (40%) patients. Pre-treatment PLT were 162 (132-205) in the group with SVR and 142 (100-191) in the group without SVR (p<0.001). Last PLT were measured 5.7 (2.1-7.6) years after SVR, at which time PLT had increased by 35 (7-62; p<0.001). In those with thrombocytopenia pre-treatment, PLT were >150 in 44 (62%) patients with SVR at last follow-up (p<0.001). In the group without SVR, the last PLT were measured after 4.4 (1.9-7.1) years and had decreased by 17 (−5-47, p<0.001). Repeated measurement analysis, including 3387 PLT measurements (interval: 0.45 [0.13-0.79] years), indicated a gradual increase in PLT following SVR and a decline in patients without SVR (p<0.001). CONCLUSION Among patients with HCV-induced advanced hepatic fibrosis the PLT gradually increase following SVR, suggesting liver histology improves with time after eradication of HCV.

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Disclosures:

Adriaan J. van der Meer - Speaking and Teaching: MSD Bart J. Veldt - Board Membership: GSK

Jordan J. Feld - Advisory Committees or Review Panels: Roche, Merck, Vertex, Gilead, Abbott, Tibotec, Theravance, Achillion; Speaking and Teaching: Merck, Roche, Abbott

Heiner Wedemeyer - Advisory Committees or Review Panels: Transgene, MSD, Roche, Gilead, Abbott, BMS, Falk; Grant/Research Support: MSD, Novartis, Gilead, Roche, Abbott; Speaking and Teaching: BMS, MSD, Novartis, IĪF

Jean-Francois Dufour - Advisory Committees or Review Panels: Bayer, BMS, Gilead, Jansse, Novartis, Roche

Michael P. Manns - Consulting: Roche, BMS, Gilead, Boehringer Ingelheim, Novartis, Idenix, Achillion, GSK, Merck/MSD, Janssen, Medgenics; Grant/Research Support: Merck/MSD, Roche, Gilead, Novartis, Boehringer Ingelheim, BMS; Speaking and Teaching: Merck/MSD, Roche, BMS, Gilead, Janssen, GSK, Novartis

Stefan Zeuzem - Consulting: Abbvie, Achillion Pharmaceuticals, Boehringer Ingelheim GmbH, Bristol-Myers Squibb Co., Gilead, Novartis Pharmaceuticals, Merck & Co., Idenix, Janssen, Roche Pharma AG, Vertex Pharmaceuticals, Presidio, Santaris, Inc

Robert J. de Knegt - Advisory Committees or Review Panels: MSD, Roche, Norgine, Janssen Cilag; Grant/Research Support: Gilead, MSD, Roche, Janssen Cilag, BMS; Speaking and Teaching: Gilead, MSD, Roche, Janssen Cilag

Harry L. Janssen - Consulting: Abbott, Bristol Myers Squibb, Debio, Gilead Sciences, Merck, Medtronic, Novartis, Roche, Santaris; Grant/Research Support: Anadys, Bristol Myers Squibb, Gilead Sciences, Innogenetics, Kirin, Merck, Medtronic, Novartis, Roche, Santaris

The following people have nothing to disclose: Raoel Maan, Frank Lammert, Andres Duarte-Rojo, Wolf P. Hofmann, Bettina E. Hansen

1471

Interferon lambda-4 genotyping to predict treatment outcome among hepatitis C patients (genotype 1 and non-1) undergoing antiviral therapy in clinical practice

Andrea Magri, Michela E. Burlone, Lisa Franzosi, Elisa Boccato, Simone Boccheffo, Rosalba Minisini, Mario Pirisi

DiMT, Universifa del Piemonte Orientale, Novara, Italy

Background and aim. On the region coding for the interferon lambda gene, a dinucleotide variant, rs67272382 (loss of T) and rs74593329 (T>G), results in a frameshift (ss469415590, TT>ΔG). In turn, TT>ΔG leads to the production of a protein, associated with poorer response to treatment in hepatitis C virus (HCV) genotype 1 patients recruited in clinical trials. Our purpose was to compare genotyping of ss469415590 with that of interleukin-28B rs12979860 as predictors of sustained virological response (SVR) in “real life”, genotype 1, 2, 3, 4 HCV patients undergoing antiviral therapy. Methods: 150 naīve HCV-infected patients (69 males, median age 53; HCV-1=75, HCV-2=50, HCV-3 = 17 and HCV-4=8; advanced fibrosis, defined as METAVIR score >F3, =64) undergoing pegylated interferon alpha and ribavirin were genotyped for both rs12979860 and ss469415590. SVR was defined as circulating HCV RNA below the limit of detectability (<25 IU/ml) six months after the end of therapy. DNA extracted from peripheral blood samples was genotyped for rs12979860 and ss469415590 by restriction fragment length polymorphism and, in 10% of samples, by sequencing. Results. There were 47 (31%) CC, 82 (55%) CT, and 21(14%) TT rs12979860 carriers; and 44 (29%) TT, 85 (57%) T/AG and 21(31%) AG/AG ss469415590 carriers. The two polymorphisms were strictly associated (weighted kappa = 0.91, 95%CI 0.85-0.97). Seventy-eight (52%) achieved SVR, 25/78 (32%) infected by HCV-and 53/78 (68%) infected with HCV-2, 3. rs12979860 CC and ss469415590 TT homozygotes were more likely to achieve a SVR either considering the entire population (32/47, 68%, p=0.015 and 31/44, 70%, p=0.012, respectively), or HCV-1 infected patients only (10/17, 59%, p=0.003 and 9/15, 60%, p=0.005, respectively). Patients with advanced fibrosis were less likely to achieve a SVR (17/64, 27% vs.61/86, 71%, p<0.001). At logistic regression, advanced fibrosis (OR 0.17, 95%CI 0.07-0.39, p<0.001), HCV genotype 2, 3 (OR 8.23, 95%CI 3.22-21.0, p<0001 and OR 3.60, 95%CI 1.04-12.5, p=0.043, respectively) and carriage of the ss469415590 TT genotype (OR 3.31, 95%CI 1.29-8.46, p=0.013) were associated with SVR, independently of age and gender. When only genotype 1 patients were considered, the two independent predictors of SVR were absence of advanced fibrosis (OR 5.65, 95%CI 1.62-19.75, p=0.007) and rs12979860 CC genotype (OR 8.61, 95%CI 2.25-33.0, p=0.002). Conclusions. In clinical practice, ss469415590 may represent the single best genetic pre-treatment predictor as far as the entire HCV population (genotype 1 and non-1) is considered, but does not replace rs12979860 to predict treatment outcome among HCV-1 infected patients.

Disclosures:

Mario Pirisi - Advisory Committees or Review Panels: Merck; Speaking and Teaching: Gilead, Bristol-Myers-Squibb

The following people have nothing to disclose: Andrea Maqri, Michela E. Burlone, Lisa Franzosi, Elisa Boccato, Simone Bocchetta, Rosalba Minisini

1472

HCV genotype 1a and 1b: similarities and differences in clinical features, therapeutic outcome and predictor of response

Filomena Morisco1, Angelo Andriulli2, Vito Si Marco3, Maurizio Margaglione4, Antonio Ippolito2, Giovanna Fattovich5, Antonina Smedile6,7, Maria R. Valvano2, Michele Milella8, Martina M. Felder9, Giovonni B. Gaeta10, Pietro Gatti11, Paolo Tundo12, Michele Barone13, Raffaele Cozzolongo14, Mario Angelico15, Giuseppe Mazzela16, Teresa Santantonio17, Rocco Granata1, Sii via Camera1, Nicola Caporaso1

1Department of Clinical Medi cine and Surgery, Federico II University of Naples, Naples, Italy; 2Division of Gastroenterology, Casa Sollievo Sofferenza Hospital, IRCCS, Son Giovanni Rotondo, Italy; 3Division of Gastroenterol ogy, University of Palermo, Palermo, Italy; 4Department of Clinical and Experimental Medicine, University of Foggia, Foggia, Italy; 5Division of Gastroenterology, University of Verona, Verona, Italy; 6Department of Medical Science, University of Torino, Torino, Italy; 7Department of Gastroenterology and Hepatology, Azienda Ospedaliera Citta della Solute e della Scienza, Torino, Italy; 8Ciinic of Infectious Diseases, University of Bari, Bari, Italy; 9Division of Gastroenterology, Central Hospital, Bolzano, Italy; 10Clinic of Infectious Diseases, University of Naples, Naples, Italy; 11DIMO Medical Oncology, University of Bari, Bari, Italy; 12Division of Infectious Diseases' S. Caterina Novella Hospital, Galatina, Italy; 13Division of Gastroenterology, University of Bari, Bari, Italy; 14Division of Gastroenterology, S. de Bellis Hospital, IRCCS, Castellana Grotte, Italy; 15Division of Gastroenterology, University of Tor Vergata, Roma, Italy; 16Division of Gastroenterology, University of Bologna, Bologna, Italy; 17Clinic of Infectious Diseases, University of Foggia, Foggia, ltaly

Aim: to evaluate the baseline similarities and differences between the two HCV-1 subtypes, 1a vs 1b, on pre-treatment of response to peg-interferon and 1184 with HCV genotype-1 infection were treated with PEG-IFN α-2a or α-2b in combination with daily ribavirin (1000-1200 mg/day). A total of 15 centers in Italy, selected by voluntary participation between January 2005 and December 2010, took part into the study. The study included 155 (13%) patients infected with subtype 1a (M/F 114/41, median age 47 yrs, range 18-78) and 1029 (87%) patients infected with subtype 1b (M/F 574/455, median age 57 yrs, range 18-84). Results: At multivariate analysis, the baseline characteristics differentiating patients with genotype 1a vs 1b were younger age (<50 yrs) and male sex, being more frequently observed in genotype 1a. Of note, the IL28B polymorphisms and the RVR resulted equally distributed between the two HCV 1 subtype. SVR was achieved by 38% of genotype 1b and by 45% of genotype 1a even in this difference of 7% is not statistically significant. At the multivariate analysis of pre-treatment and on-treatment predictors of SVR, three factors were independently associated in subtype 1a: female gender (OR = 2.829, Cl: 1.146-6.983), IL28B polymorphism (OR = 5.216, Cl: 1.765-15.410), and RVR (OR =5.066, Cl: 1.926-13.328); and three factors independently associated in subtype 1b: IL28B polymorphism (OR = 3.303, Cl: 2.256 −4.834), RVR (OR = 7.139, Cl: 4.721-10.796), and low baseline serum HCV-RNA concentration (< 400.000 IU/ mL)(OR = 2.123, Cl: 1.474-3.059). In subtype 1b the RVR status emerges as the most predictive characteristic of SVR, its strength outweighing the power of IL28 polymorphisms; on the contrary, in subtype 1a the two previous predictors appear to have an identical efficiency in predicting SVR. Conclusion: Our study conducted in a large nation-wide cohort of naīve patients with HCV subtype 1a and 1b infections shows that genotype 1a are more frequently observed in young male patients. The study also pinpoints some differential predictive features of SVR between the two subtypes, a finding which would imply that the two subtypes be separately evaluated in future therapeutic trials.

Disclosures:

Giovanni B. Gaeta - Advisory Committees or Review Panels: Janssen, Merk, BMS, Novartis, Gilead, Roche, Janssen, Merk, BMS, Novartis, Gilead, Roche, Janssen, Merk, BMS, Novartis, Gilead, Roche, Janssen, Merk, BMS, Novartis, Gilead, Roche

Mario Angelico - Advisory Committees or Review Panels: Gilead; Grant/Research Support: Roche; Speaking and Teaching: GSK, Roche, Gilead, Novartis

The following people have nothing to disclose: Filomena Morisco, Angelo Andriulli, Vito Di Marco, Maurizio Margaglione, Antonio Ippolito, Giovanna Fattovich, Antonina Smedile, Maria R. Valvano, Michele Milella, Martina M. Felder, Pietro Gatti, Paolo Tundo, Michele Barone, Raffaele Cozzolongo, Giuseppe Mazzella, Teresa Santantonio, Rocco Granata, Silvia Camera, Nicola Caporaso

1473

Hepatitis C Testing is Low in a Large Primary Care Center

Euriko Torrazza Perez, Mohamed G. Shoreibah, Brendan M. McGuire, Joseph R. Bloomer, Omor Massoud

Gastroenterology and Hepatology, University of Alabama at Birmingham, Birmingham, AL

BACKGROUND: Hepatitis C virus (HCV) is the most common blood-borne infection in the United States is with an estimated prevalence of 2.7-3.9 million persons (1.0-1.5%). Although, it is a curable disease, it is under-diagnosed and under-treated. Little is known about healthcare providers adherence to guidelines recommending testing populations at higher risk for HCV. PURPOSE: To examine testing practices among primary care physicians and determine which patient variables are associated with testing for hepatitis C virus antibody (anti-HCV) and HCV infection. METHODS: Participants for this cross-sectional study were from the Birth-Cohort Evaluation to Advance Screening and Testing for Hepatitis C (BEST-C-I) study. Electronic medical records for patients seen in the primary care clinics at the University of Alabama at Birmingham (Kirklin Clinic) between 1/1/2005 and 12/31/2010 were reviewed. A multivariable logistic regression analysis was conducted to calculate odds ratios (ORs) and 95% confidence intervals (CI) for predictors of being tested for hepatitis C and, in a separate model, to test positive for anti-HCV. RESULTS: Of 39, 240 participants, 1, 677 (4.3%) were tested for HCV and 72 (0.2%) were positive. Of 5, 355 participants with any documented risk factors, 1, 073 (20%) were tested for anti-HCV. Only 1% of the medical records had documentation regarding illicit drug use and/or blood transfusions. Participants born from 1945 through 1965 accounted for 72.2% of all anti-HCV+ persons. In a multivariable analysis, males (OR= 1.80, 95% Cl 1.62-2.00), HIV infected individuals (OR= 31.79, 95%CI 19.30-52.39), endstage renal disease patients (OR= 28.11, 95%CI 14.9952.69), hemophiliacs (OR= 5.72, 95% Cl 3.83-8.54) patients with elevated alanine transaminase (ALT) (OR= 10.21, 95%CI 8.87-11.76) were more likely to be tested for anti-HCV. Participants born between 1945 and 1965 were more likely (OR 9.73, 95% Cl 4.32-21.93) to be anti-HCV positive. CONCLUSION: Testing for anti-HCV was low in a large primary care center. Many persons at risk for HCV infection remained untested. Strategies for improving anti-HCV screening in primary care settings are recommended for patients at increased risk. The increased likelihood anti-HCV positivity among those born between 1945 and 1965 supports the new Centers for Disease Control and Prevention (CDC) recommendation of one time testing for all individuals in this cohort group.

Disclosures:

Brendan M. McGuire - Consulting: hepahope; Grant/Research Support: roche (genentech) laboratories, bayer healthcare, vital therapies, ikaria therapeutics, cumberland pharmaceuticals, vertex pharmaceuticals

The following people have nothing to disclose: Euriko Torrazza Perez, Mohamed G. Shoreibah, Joseph R. Bloomer, Omar Massoud

1474

Enhanced Liver Fibrosis (ELF) Panel as a Predictor of Liver Fibrosis in Chronic Hepatitis C Patients

Flavia F. Fernandes1 , 2, Alessondro Dellavance3, Luis Eduardo C. Andrade3, Frederico F. Compos1 , 2, Gustavo Pereiro2, Carlos Terra1, João Luiz Pereiro2, Fátimo A. Figueiredo1, Renata M. Perez1, Maria Lucia G. Ferraz3; 1Hospital Universitário Pedro Ernesto, Rio de JAneiro, Brazil; 2Hospital Federal de Bonsucesso, Rio de JAneiro, Brazil; 3Fleury LAborofory, São Poulo, Brazil

Fibrosis is the most important issue in the assessment of chronic hepatitis C (CHC). Liver biopsy (LB) remains the reference for staging fibrosis. Transient elastography (TE - Fibroscan®) is well-established as a non-invasive exam to evaluate fibrosis in CHC. ELF is a promising non-invasive serological marker panel which comprises hyaluronic acid (HA), tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) and aminoterminal propeptide of procollagen type III (PIIINP). To evaluate the performance of ELF in CHC patients, we evaluated 120 patients with CHC submitted to LB and TE. Exclusion criteria: HIV and HBV co-infection, daily alcohol intake of more than 40g, cholestasis, chronic kidney failure, right-sided heart failure, fibrogenic drugs use, less than 6 portal tracts or concomitant pathology in the liver biopsy. After LB, TE was performed and a blood sample collected in a three months' time. Serum was frozen at - 70°. ELF score was calculated using the algorithm: ELF = 2.278 + 0.851 ln(HA) + 0.751 ln(PIIINP) + 0.394 ln(TIMP-1). Cut-off points proposed by the manufacturer were applied: < 7.7 absent or mild fibrosis, > 7.7 and < 9.8 moderate fibrosis and > 9.8 severe fibrosis. TE results were expressed in kilopascals (kPa) and the median value of 10 acquisitions was considered for analysis. Only exams with a success rate of at least 60% and an IQR/M ratio of 30% were considered. LB was reviewed by one experienced pathologist. The study was approved by the local Ethics Committee. SPSS 17.0 (SPSS Inc., Chicago IL) was used for statistical analyses. Results: 34% of the patients were men, mean age 53 (SD ± 11.3) years old. The distribution of fibrosis stages according to METAVIR was: stage 0 - 2%, stage 52%, stage 2 - 30%, stage 3 - 9% and stage 4 - 7%. According to ELF cut-off points we had: three (2%) patients with absent or mild fibrosis (F0-1), seventy-four (61%) with moderate fibrosis (F2-3) and forty-four (37%) with cirrhosis (F4). ELF overestimated fibrosis in 76% of cases and underestimated in one case. The ELF accuracy (AUROC) for the significant fibrosis (F>2) was 0.81 (95% IC: 0.73-0.87), for advanced fibrosis (f≥3) was 0.82 (95% IC: 0.74-0.88) and cirrhosis was 0.78 (95% IC: 0.70-0.85). We found no statistical significant difference when comparing the AUROCs of ELF and TE for significant fibrosis (p 0.13), advanced fibrosis (p 0.08) and cirrhosis (p 0.11) Conclusion: ELF panel had a good performance as a non-invasive marker. However, new cut-off points need to be established to improve the discrimination of different stages of fibrosis in CHC patients.

Disclosures:

The following people have nothing to disclose: Flavia F. Fernandes, Alessandra Dellavance, Luis Eduardo C. Andrade, Frederico F. Campos, Gustavo Pereira, Carlos Terra, Joao Luiz Pereira, Fatima A. Figueiredo, Renata M. Perez, Maria Lucia G. Ferraz

1475

Hepatitis C Virus Genotype 2 may not be detected by the Cobas AmpliPrep/Cobas TaqMan HCV Test, version 1.0

Tsunamasa Watanabe1,Takako Inoue1, Yasushi Tanoue2, Hisato Maekawa2, Etsuko lio1, Kayoko Matsunami1, Noboru Shinkai1, Makoto Yoshiba2, Yasuhito Tanaka1

1 Nagoya City University, Nagoya, Japan; 2Sempo Takanawa Hospital, Tokyo, Japan

Hepatitis C viremia is diagnosed by detecting hepatitis C virus (HCV) RNA using real-time polymerase chain reaction (PCR)based assays. Two highly sensitive commercial assays for HCV RNA quantification are available in many countries: the Roche Cobas AmpliPrep/Cobas TaqMan HCV assay (CAP/CTM HCV) and the Abbott RealTime HCV assay (ART HCV). Despite its good performance with most HCV strains, the CAP/CTM HCV test, version 1.0 (v1.0) fails to detect the certain genotype 2 strains with single nucleotide polymorphisms at positions 145 and 165 in the 5' untranslated region (5' UTR). We report two Japanese patients with HCV genotype 2a in whom HCV RNA was undetectable by CAP/CTM HCV v1.0, although viremia was confirmed by the ART HCV test (4.2 and 4.0 Log10 IU of HCV RNA/mL) and Architect HCV Core Antigen assay. This failure could be related to two or three substitutions in the putative binding site for the TaqMan probe. The substitutions are at positions 145, as described for HCV genotype 4, and at the other positions, which have not been reported previously. Underestimation of HCV genotype 2 RNA by CAP/CTM HCV v1.0 has been reported previously but failure to detect HCV genotype 2a RNA is critical because this is the second most common HCV genotype. Recently, a second version of the assay, CAP/CTM HCV v2.0, with redesigned primers and an additional probe, has been released in Western Europe and the USA and the problem of underestimating the quantity of certain genotype 4 HCV strains has been reported to have been solved. CAP/CTM HCV v2.0 could detect HCV genotype 2a RNA in the two samples missed by the v1.0 assay but clinicians who use the CAP/CTM HCV v1.0 assay routinely should be aware of the potential for false negative results.

2 patients infected with HCV in whom the CAP/CTM HCV v1.0 assay failed to detect HCV RNA

CAP/CTM HCV vl. O (Log IU/mL)HCV Core Ag (fmol/L)ART (Log IU/mL)Genotype
case 1target not detected97.14.22a
case 2target not detected12.64.02a

Disclosures:

Yasuhito Tanaka - Advisory Committees or Review Panels: Nippon Boehringer Ingelheim Co., Ltd.; Grant/Research Support: Chugai Pharmaceutical CO., LTD., MSD, Mitsubishi Tanabe Pharma Corporation, Dainippon Sumitomo Pharma Co., Ltd., DAIICHI SANKYO COMPANY, LIMITED, Bristol-Myers Squibb

The following people have nothing to disclose: Tsunamasa Watanabe, Takako Inoue, Yasushi Tanoue, Hisato Maekawa, Etsuko lio, Kayoko Matsunami, Noboru Shinkai, Makoto Yoshiba

1476

Point of Care Hepatitis C Screening During Endoscopy

Raja Taunk, Daniel I. Zapata, Jennifer Stone, Victoria Menashy, Ilan Weisberg

Lenox Hill Hospital, New York City, NY

Objective: Chronic hepatitis C viral (HCV) infection remains a largely undiagnosed chronic disease process. It is estimated that 75% of patients infected with hepatitis C are unaware they have it. The Centers for Disease Control recently released guidelines advising birth cohort screening for people born between 1945-1965. We performed a novel prospective trial to evaluate the efficacy, efficiency, and patient acceptance of point of care (POC) hepatitis C testing during endoscopy and colonoscopy Methods: At a private practice endoscopy center in Manhattan, 254 consecutive patients born between 19451-965 who were scheduled for colonoscopy or upper endoscopy for any reason were offered point of care testing for hepatitis C. OraQuick Rapid HCV antibody test was utilized to provide point of care HCV screening with results available in 20-40 minutes. All patients completed a questionnaire prior to endoscopy regarding risk factors and exposures. Patients who tested positive for HCV were contacted for further testing and possible therapy. Results: HCV screening was offered to 254 patients and 220 agreed to be tested. Thirty-four patients declined testing (14%). The majority (62%) who declined were women. Two patients were found to be hepatitis C positive (1%); however both were already known to have hepatitis C. No new diagnoses of chronic hepatitis were made. Traditional risk factors for HCV transmission were prevalent, including illicit drug use (21.4%), acupuncture (19.5%), STD's (12.3%), tattoos (8.2%), and IVDU (2.3%). There was a Caucasian predominance to the cohort of 62%. Point of care testing and counseling did not affect the work-flow or efficiency of the endoscopy center. Prior to initiation of this study, average patient time after the nurse's interview to patient discharge was 1 hour 52 minutes. During the enrollment and HCV testing period the patient time to proceed through endoscopy was largely unchanged at 1 hour 47 minutes. Conclusions This prospective study demonstrates that it is possible to test patients for viral hepatitis during routine colonoscopy and endoscopy in an efficient and time effective manner. Earlier studies documented a 1% new diagnosis rate, however despite prevalent traditional HCV risk factors in our study cohort we had no new diagnoses at this interval analysis. Point of care testing allows for high rates of patient acceptance. Also, testing did not delay workflow at the endoscopy center. Given recent CDC recommendations for birth cohort screening for HCV, gastroenterologists should strongly consider PoC testing in non-traditional patient interactions such as endoscopy.

Disclosures:

The following people have nothing to disclose: Raja Taunk, Daniel I. Zapata, Jennifer Stone, Victoria Menashy, llan Weisberg

1477

Comparison of the Enhanced Liver Fibrosis (ELF) score and transient elastography in haemophilia patients with chronic hepatitis C

Greet Boland, Lisa Manen van, Evelien Mauser-Bunschoten, Dietje Fransen-van de Putte, Karel J. van Erpecum

University medical center Utrecht, Utrecht, Netherlands

Introduction: Progressive liver damage due to hepatitis C virus infection is a major cause of co-morbidity in haemophilia patients. The need for accurate diagnostic tools for assessing the extent of liver fibrosis is high in these patients. The aim of this study was to investigate the additional value of the enhanced liver fibrosis (ELF) test combining hyaluronic acid, procollagen-III-amino terminal peptide and tissue inhibitor of metalloproteinase-1 compared to transient elastography (TE). Methods: We evaluated the ELF score in 58 HCV-infected haemophilia patients on the ADVIA Centaur XP (Siemens). TE was performed by using the FibroScan (Echosens, Paris, France). In 15 patients (25.9%), TE measurement did not meet manufacturers' conditions for a reliable result. ROC curves were made and cut-off values calculated. Results: The ELF-test results were compared with the outcome of TE. AUROC for severe fibrosis (>F3) was 0.876 (95% C. l.0.757-0.995). AUROC for significant fibrosis (>F2) was 0.732 (95% C. I. 0.597-0.866). The cut-off value of 10.3 of the ELF-score based on ROC curve predicted severe fibrosis with a sensitivity of 66.7%, a specificity of 95.3%, NPV of 89.1% and PPV of 83.3%. For exclusion of fibrosis, a cut-off value of 7.7 yielded an NPV of 88.9%. For diagnosing significant fibrosis, a cut-off value of 10.2 yielded a PPV of 85.7%. Conclusion: The ELF test is a good discriminator for severe fibrosis and can exclude fibrosis with a high certainty in haemophilia patients with hepatitis C.

Disclosures:

Karel J. van Erpecum - Grant/Research Support: Bristol Meyers Squibb, MSD

The following people have nothing to disclose: Greet Boland, Lisa Manen van, Evelien Mauser-Bunschoten, Dietje Fransen-van de Putte

1478

The prevalence of resistance mutations to HCV Direct acting antiviral agents in Chinese treatment-naϊve patients

Cai Qing-Xian, Liu Ying, Zhao Zhixin

The third affiliated hospifol of Sun yat-sen University, Guangzhou, China

Background: The emergence of direct acting antiviral agents(DAAs) had brought about great changes to the treatment of chronic hepatitis C. However, gene polymorphism of HCV and high viral variability would naturally cause resistance to the DAAs. In this study, we tried to detect natural polymorphisms and illustrate the prevalence of such mutations in Chinese treatment-naīve patients. Methods: A total of 184 treatment-naīve chronic hepatitis C patients from the third affiliated hospital of Sun Yat-sen University were enrolled. HCV genotypes were determined by direct sequencing and phylogenetic tree analysis based on HCV core and NS5B conserved regions sequence. Several nested PCR assays with genotypespecific primers were performed to amplify the HCV viral regions of NS3, NS5A and NS5B. Results: The genotyping result showed that 1 84 patients were classified into 3 categories: genotype 1b, 2a and 6a at frequencies of 40.2%(74/184), 8.2%(15/184), 51.6%(95/184). We also successfully amplified 88.04%(162/184) in NS3, 86.96%(160/184) in NS5A as well as 84.24%(155/184) in NS5B. For NS3 sequences, a total of 266 amino acid substitutions were detected in 125 (77.16%) patients. Major resistant-mutation A156S was found in 18.33% of patients with HCV 1b and 64.28% of patients with HCV 2a, while Q80K and V170I variability were detected in 95.45% and 100% of HCV 6a. None of the 162 individuals had the substitution V55A and R155K/T/Q. The proportion of these 3 resistance mutations (Q80K, A156S, V170I) in different groups were obviously different(p<0.05). For NS5A sequences, resistant-mutations Q30R was detected in 116 cases of HCV 1b and 6a, while L31M was found 12 of HCV 2a and 4 of HCV 6a, H58P was discovered in 42.5%(68/160) patients with the above genotypes, Y93C was showed in 9 individuals only with genotype 2a. For NS5B sequences, C316N were detected among all HCV 1b patients, while s282T was found in 20.73% (17/82) of HCV 6a. Conclusions: Naturally occurring dominant resistance mutations to HCV DAAs do pre-exist in Chinese treatmentnaīve patients. HCV 2a and 6a have more major resistant-mutation than HCV type 1b.

Disclosures:

The following people have nothing to disclose: Cai Qing-Xian, Liu Ying, Zhao Zhixin

1479

Feasibility and Accuracy of Elastpq® Shear Wave Elastography Technique and Doppler Indices in the Noninvasive Assessement of Liver Fibrosis: A Preliminary Experience

Matteo Garcovich, Maria Assunta Zocco, Laura Riccardi, Davide Roccarina, Brigida E. Annicchiarico, Maria Elena Ainora, Gianluigi Caracciolo, Francesca Romana Ponziani, Antonio Grieco, Massimo Siciliano, Maurizio Pompili, Antonio Gasbarrini

Catholic University of Sacred Heart, Rome, Italy

Introduction&Aims: Real-time shear wave elastography (RTE) is a novel non-invasive technique that assesses liver fibrosis by measuring liver stiffness (in kPa). The purpose of this study was to determine the efficacy and the feasibility for the assessment of hepatic fibrosis as compared with the histological grade of fibrosis in patients undergoing liver biopsy. Methods: Consecutive patients scheduled for liver biopsy were studied by using the iU22 ultrasound system (Philips Medical Systems) with a convex probe and ElastPQ technique (performed both on the right and left lobe of the liver). In addition, Doppler indices inclusive of resistive and pulsatility index at various sites, hepatic vein (HV) and portal venous blood velocity and flows (including damping index) were evaluated. The correlations between these quantitative parameters and the pathological findings (Metavir score) were analyzed using Spearman rank correlation coefficients and receiver operating characteristic curve analyses were performed to calculate area under the curve (AUC) for F>2, F>3, and F=4. Results: We enrolled 45 patients (28 males and 17 females) who underwent ultrasoundguided liver biopsy for viral or non-viral chronic hepatitis (HCV −60%; NASH - 33%). Liver stiffness measurement performed on the right lobe were reliable in all cases, while in 11% of patients left lobe elastography was not obtainable or unreliable. Median values were 4.11 (range 3.23-4.41) kPa and 3.67 (2.51-6.73) kPa for F0-F1, 7.1(4.28-12.9) kPa and 8.38 (5.85-12.3) kPa for F2-F3, 13.58 (9.9-20.79) and 19.98 (10.31-31, 34) kPa for F4 in the right and left lobe, respectively. AUCs calculated for the right lobe were 0.91 (0.85-0.92; 95%CI) for F>2, 0.88 (0.73-0.90; 95%CI) for F>3 and 0.96 (0.91-0.98; 95%CI) for F=4. As concerning Doppler measurements, only damping index correlated slightly with the grade of fibrosis, while adding Doppler indices to liver stiffness increased no further the diagnostic accuracy of RTE. Conclusion: RTE with ElastPQ appears to be a useful tool for non-invasive evaluation of fibrosis in patients with viral and non-viral chronic hepatitis, although these findings need to be confirmed in larger studies.

Disclosures:

The following people have nothing to disclose: Matteo Garcovich, Maria Assunta Zocco, Laura Riccardi, Davide Roccarina, Brigida E. Annicchiarico, Maria Elena Ainora, Gianluigi Caracciolo, Francesca Romana Ponziani, Antonio Grieco, Massimo Siciliano, Maurizio Pompili, Antonio Gasbarrini

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