Adaptive Immunity


A novel approach to ameliorate liver inflammation and disease progression in a murine model of chronic liver disease

Nadine Hermanns1, Francisco Javier Cubero1, Antje Mohs1, Kim Ohl2, Malika Al Masaoudi1, Christian Liedtke1, Klaus Tenbrock2, Christian Trautwein1;

1 Department of Medicine III, RWTH University Hospital, Aachen, Germany;2Department of Pediatrics, Division of Allergology and Immunology, RWTH University Hospital, Aachen, Germany

Background & Aim: The Interleukin-17 (IL-17)-mediated immune response has been shown to play a crucial role in inflammation-associated disease. Actually, serum IL-17 levels have been incorporated in the clinic as a marker of severity of liver injury. Since, the transcription factor cAMP-responsive element modulator (CREM)-α contributes to increased IL-17 expression observed in patients with liver disease, we reasoned whether overexpression of CREM-α has an impact on the initiation and progression of liver fibrosis and hepatocellular carcinoma (HCC). Methods: Transgenic mice overexpressing CREM-α in T-Cells (controlled by CD2 promotor) were crossed with hepato-cyte-specific Nemo knockout (NemoΔhepa) mice, a model of chronic liver injury. Here, the phenotype was characterized during the progression of acute and chronic liver injury. Results: In 8 week old NemoΔhepa/CREM-α compared with NemoΔhepa mice, we found significantly reduced serum AST and ALT levels. These findings were associated with lower absolute numbers of infiltrating CD11 b+ and F4/80 cells in NemoΔhepa /CREM-α livers. In addition, we found significantly elevated mRNA expression levels of cytokines IL-10 and IL-4 in both T-cells and liver tissue in NemoΔhepa/CREM-α compared with NemoΔhepa and WT mice suggesting that the CREM-α transgene in T-cells influences liver inflammation towards a protective environment. Liver histology and sirius red staining revealed that fibrosis was significantly reduced in NemoΔhepa/CREM-α compared to NemoΔhepa livers in 13 weeks old animals. This was further confirmed by studying extracellular matrix deposition showing significantly reduced Collagen IA1 and fiber deposition in NemoΔhepa/CREM-α livers, which was accompanied by decreased desmin-associated activation of Hepatic Stellate cells. In 52 weeks old NemoΔhepa/CREM-α livers, a significantly reduced liver-body-weight ratio and significantly less nodules in comparison to

NemoΔhepa mice were evident. Additionally, c-myc mRNA levels and protein levels of glutamine synthetase revealed lower cancer related-metabolism in NemoΔhepa/CREM-α livers. Conclusion: Our results demonstrate that overexpression of CREM-a in T-cells in NemoΔhepa mice attenuates disease progression as shown by reduced liver fibrosis and growth of HCC. This finding is the result of changing inflammation in these livers towards a protective milieu by enhancing the expression of distinct cytokines (IL-4, IL-10) and by reducing immune cell infiltration and IL-17 production. The present findings suggest a new molecular approach to reduce disease progression in chronic liver diseases.


Christian Trautwein - Grant/Research Support: BMS, Novartis, BMS, Novartis; Speaking and Teaching: Roche, BMS, Roche, BMS

The following people have nothing to disclose: Nadine Hermanns, Francisco Javier Cubero, Antje Mohs, Kim Ohl, Malika Al Masaoudi, Christian Liedtke, Klaus Tenbrock


Antibody-Secreting Cells with a Phenotype of Ki-67low, CD138high, CD31high and CD38high Secrete Non-Specific IgM during Primary Hepatitis A Virus Infection

Hyun Woong Lee1, Seokchan Hong2, Dong-Yeop Chang2, Hyung J. Kim1, Eui-Cheol Shin2;

1 Department of Internal Medicine, Chung-Ang University College of Medicine, Seoul, Republic of Korea; 2Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon, Republic of Korea

Background/Aims: We investigated the nature of ASCs here by direct ex vivo assays in patients with acute hepatitis A (AHA) which is caused by the primary infection of hepatitis A virus (HAV). Methods : The study included 39 patients diagnosed with AHA infection who were hospitalized at Chung-Ang University Hospital. All patients were seropositive for anti-HAV IgM, and all had clinical features of acute hepatitis. Peripheral blood samples at the acute stage were collected on the day of admission from all of the 39 patients. Follow-up sampling was performed at the subacute stage (5–14 days) or at the convalescent stage (35–150 days). Results : A robust plasmablast response was detected in peripheral blood during the acute stage and was dominated by IgM secretion. It was demonstrated that a substantial portion of the response was non-virus-specific in the study of the plasmablasts and the secreted IgM. We detected HAV-specific plasmablasts by staining with fluorochrome-tagged VP1 protein and compared them with non-HAV-specific plasmablasts. Non-HAV-specific plasmablasts have the phenotype of Ki-67low/CD138high/CD31high/CD38high as compared with HAV-specific plasmablasts, demonstrating that non-HAV-specific plasmablasts have a bone marrow (BM) plasma cell-like phenotype while HAV-specific plasmablasts have a typical phenotype of circulating plasmablasts. Conclusions : These data suggest that non-HAV-specific plasmablasts are mobilized ASCs from the BM niches of plasma cells, whereas HAV-specific plasmablasts are newly generated ASCs. In this study, we demonstrated that pre-existing BM plasma cells are released to circulation during AHA and contribute to the non-virus-specific ASC response and IgM secretion.


Phenotypes of HAV-specific and non-HAV-specific ASCs in the peripheral blood


The following people have nothing to disclose: Hyun Woong Lee, Seokchan Hong, Dong-Yeop Chang, Hyung J. Kim, Eui-Cheol Shin


Cellular immune responses for human homologue of Prp24p-derived peptides in patients with Hepatocellular Carcinoma

Kiichiro Kaji, Eishiro Mizukoshi, Tatsuya Yamashita, Kuniaki Arai, Hajime Sunagozaka, Kazumi Fushimi, Hidetoshi Nakagawa, Kazutoshi Yamada, Masaaki Kitahara, Shuichi Kaneko;

Gastroen-terology, Graduate School of Medicine, Kanazawa University, Kanazawa, Japan

Background/Aim: Human homologue of Prp24p is an RNA-binding nuclear protein and also known as squamous cell carcinoma antigen recognized by T cells (SART3). It is expressed in many malignant tumor cell lines and function as tumor rejection antigens (TRA). In addition, peptides containing SART3 epi-topes are capable of generating cytotoxic T cells (CTLs), and therefore, have been used for immunotherapy to treat several kinds of cancers. In this study, we examined human homologue of Prp24p expression in various hepatoma cell lines and HCC tissues of patients, and analyzed immune responses to this molecule using peripheral blood mononuclear cells (PBMCs) and tumor-infiltrating lymphocytes (TILs) to investigate the usefulness of this molecule as an immunotherapeutic target in hepatocellular carcinoma (HCC). Methods: The expression of human homologue of Prp24p in hepatoma cell lines and HCC tissues was confirmed by immunofluorescence and immunohistochem-ical analysis. Two peptides derived from human homologue of Prp24p were synthesized (SART3–109 and SART3–315). CTL responses were investigated by interferon gamma enzyme-linked immunospot (ELISPOT) and CTL assays using PBMCs and TILs in 9 healthy donors and 49 patients with HCC. The safety of immunotherapy using human homologue of Prp24p-derived peptide was investigated by s.c. vaccinations of the peptide (SART3–109) to 12 patients with HCC (trial registration: UMIN000005677). Results: Immunofluorescence and immuno-histochemical analysis showed human homologue of Prp24p to be expressed in 7 HCC cell lines and in HCC tissue including alpha-fetoprotein (AFP)-negative individuals. Human homologue of Prp24p-specific CTLs were generated by stimulating PBMCs of HCC patients with the peptide derived from this molecule, and they showed cytotoxicity against HCC cells expressing the protein. The frequency of CTLs specific for SART3–109 and SART3–315 investigated by ELISPOT assay was 5.5±11.4 and 6.1 ±8.8 /3x105 PBMCs in HCC patients, respectively. The infiltration of SART3–109-specific interferon-gamma-pro-ducing CTLs into the tumor site was confirmed. In the vaccination study, no severe adverse events were observed and the peptide-specific CTLs were induced in 4 of 12 patients tested. Conclusions: Human homologue of Prp24p is an immunotherapeutic candidate and the peptides derived from this antigen could be useful for HCC immunotherapy.


Shuichi Kaneko - Grant/Research Support: MDS, Co., Inc, Chugai Pharma., Co., Inc, Toray Co., Inc, Daiichi Sankyo., Co., Inc, Dainippon Sumitomo, Co., Inc, Aji-nomoto Co., Inc, MDS, Co., Inc, Chugai Pharma., Co., Inc, Toray Co., Inc, Daiichi Sankyo., Co., Inc, Dainippon Sumitomo, Co., Inc, Ajinomoto Co., Inc, Bayer Japan

The following people have nothing to disclose: Kiichiro Kaji, Eishiro Mizukoshi, Tatsuya Yamashita, Kuniaki Arai, Hajime Sunagozaka, Kazumi Fushimi, Hidetoshi Nakagawa, Kazutoshi Yamada, Masaaki Kitahara


Gene-Expression Profiling to Predict Outcome of Infection in Chimpanzees Vaccinated Against Hepatitis C Virus (HCV)

Hongying Duan1, Iryna Zubkova1, Youkyung Choi2, Frances Wells1, Esther Chang3, Kathleen F. Pirollo3, Kris Krawczynski2, Robert Lanford4, Stephen Feinstone1, Marian E. Major1;

1CBER, FDA, Bethesda, MD; 2Division of Viral Hepatitis, CDC, Atlanta, GA; 3Dept of Oncology, Georgetown University, Washington, DC, DC; 4Dept of Virology and Immunology, texas Biomedical Research Institute, San Antonio, TX

Any successful HCV vaccine must induce effective immune responses that are able to control viral replication and lead to rapid clearance. It has been previously established that chimpanzees that clear primary HCV infections develop memory CD4+ and CD8+ T-cell responses that can rapidly clear secondary infections. To further characterize the intrahepatic immune response profile important for control and clearance of HCV infections, we used a Taqman Low Density immune panel consisting of 96 human immune response genes to analyze the profile of immune-related gene regulation in liver biopsy tissue before and after HCV challenge in 4 vaccinated and 3 rechal-lenged chimpanzees. All 3 rechallenged animals cleared HCV within 4 weeks while different outcomes were observed in the vaccinated animals: clearance; transient control or persistence. Following challenge, all the animals that cleared the virus showed up-regulation of a greater number of the immune related genes at a higher frequency compared with the animals that developed a persistent infection. After clearance of the virus, the expression of these genes in the immune panel decreased to baseline, prechallenge levels. IFN-gamma, CXCL10 and CXCL11 were up-regulated in all animals after challenge regardless of the outcome of the infection. Up-regulation of CD4, CD34, CD40 (T cell differentiation, activation and signaling related genes) and C3 (innate immunity related gene) were correlated with the clearance of virus in vaccinated and rechallenged animals. CCL19 (cytokine related gene) and CSF1 were shown to be associated with viral clearance in control, vaccinated and rechallenged animals. Up-regulation of CD80 (T cell receptor), SKI( cytokine) and BCL2 (apoptosis related gene) were shown to be associated with viral clearance only in the rechallenged animals. This study indicates that the intrahepatic immune responses involved in the clearance of HCV are different between animals in which the immune system has been primed by vaccination and rechallenged animals where the immune system has been primed by natural infection with HCV. Low density arrays may be a useful method to select immune response markers to predict the outcome of HCV infection or the success of a vaccine.


Esther Chang - Consulting: SynerGene Therapeutics, Inc.

Kathleen F. Pirollo - Grant/Research Support: SynerGene Therapeutics, Inc

Stephen Feinstone - Independent Contractor: Dynavax

The following people have nothing to disclose: Hongying Duan, Iryna Zubkova, Youkyung Choi, Frances Wells, Kris Krawczynski, Robert Lanford, Marian E. Major


An appropriate amount of Sorafenib could suppress the myeloid derived suppressor cells (MDSC) and Tregs in HCC patients

Yasuteru Kondo, Tomoaki Iwata, Osamu Kimura, Masashi Ninomiya, Tatsuki Morosawa, Eiji Kakazu, Takayuki Kogure, Tooru Shimosegawa;

Gastroenterology, Tohoku University Hospital, Sendai, Japan

[Background] It has been reported that MDSC and Tregs were major suppressors of the immune response against Hepatocel-lular carcinoma (HCC). Sorafenib, an oral multi-kinase inhibitor, has been approved for the treatment of HCC. Sorafenib could inhibit the MAPK and VEGF signaling. VEGF signaling might affect MDSC development as well as angio-genesis. [Aim] The aim of this study is to analyze whether sorafenib could suppress MDSC and Tregs development in HCC patients ex vivo and in vitro. [Methods] ex vivo analysis: Thirty-five HCC patients who received sorafenib were enrolled in this study. Sorafenib exhibits inter-individual pharmacokinetic variability based on the activity of CYP3A4. Therefore, we quantitated the sorafenib and sorafenib N-oxide in serum by an optimized HPLC-UV led method. The linear range of detection was 0.03–30 μg/ml. Peripheral blood mononuclear cells (PBMCs) were used for the analysis of MDSCs, Tregs and Th1. PBMCs were stained with CD3, CD4, CD25, CD127, CCR5, CXCR3, CD11 b, CD14, CD16, CD33, PD-L1, and HLA-DR antibody and analyzed by FACS canto-II. IL10 or IFN-γ secreting cells were analyzed by cytokine secreting assay. The mRNA expression of PBMCs was analyzed by deep sequence analysis (Transcriptome analysis) and realtime-PCR analysis (GM-CSF, IFN-γ,IL10, TGF-β1, arginase 1, iNOS, PD-L1). in vitro analysis: Isolated PBMCs were used to analyze the induction of MDSC and Tregs by the soluble factor induced from various hepatoma cell lines (Hep3B, Li3, PLC etc.) in a 0.4μm pore tran-swell system. NOG mice were used for the transplantation of HCC with MDSC. [Results] ex-vivo: The frequency of MDSC in HCC patients was significantly higher than those in healthy subjects. The frequencies of PD-L1 high MDSCs and Tregs were significantly decreased after 8 weeks sorafenib treatment (p<0.01). On the other hand, the frequency of Th1 cells and the ability of IFN-γ secretion in T cells were significantly increased after 8 weeks sorafenib treatment (p<0.01). The expression of GM-CSF mRNA was significantly decreased after 8 weeks sorafenib treatment (p<0.05). The concentration of sorafenib at 8 weeks sorafenib treatment and the area under the blood concentration time could be significantly correlated with the change of MDSC and Tregs frequencies(p<0.001). in vitro: Co-culture of hepatoma cell lines with PBMCs could induce MDSC and Tregs. However, 2μg/ml sorafenib not 0.5μg/ml sorafenib could suppress the induction of MDSC. [Conclusion] An appropriate amount of sorafenib could suppress the MDSC and Tregs in HCC patients and the induction of MDSC and Tregs in the in vitro model of HCC microenvironment.


The following people have nothing to disclose: Yasuteru Kondo, Tomoaki Iwata, Osamu Kimura, Masashi Ninomiya, Tatsuki Morosawa, Eiji Kakazu, Takayuki Kogure, Tooru Shimosegawa


Therapeutic Efficacy and Safety of OK432-Stimulated Monocyte-Derived Dendritic Cell Injection into Hepato-cellular Carcinoma after Radiofrequency Ablation

Masaaki Kitahara, Eishiro Mizukoshi, Kiichiro Kaji, Kazutoshi Yamada, Hidetoshi Nakagawa, Hajime Sunagozaka, Kuniaki Arai, Tatsuya Yamashita, Shuichi Kaneko;

Kanazawa Univercity, Kanazawa, Japan

BACKGROUND: Dendritic cell (DC)-based immunotherapies are believed to contribute to the eradication of the residual and recurrent tumor cells including hepatocellular carcinoma (HCC). We have developed the combined therapy of transcatheter hepatic arterial embolization (TAE) with infusion of OK432, a Streptococcus-derived anticancer immunotherapeutic agent, stimulated monocyte-derived DCs (MoDCs) for HCC, and indicated that patients treated with TAE and OK432-stimulated DC transfer had prolonged recurrence-free survival compared with the historical controls that had been treated with TAE alone (Clin.Exp.Immunol. 163:165,2011). Based on the results, the present study was designed to assess the safety, bioactivity and clinical response of OK432-stimulated MoDC infusion into HCC following radiofrequency ablation (RFA), which is more radical and curative treatment. METHODS: MoDCs were derived from peripheral blood monocytes of hepatitis C-related HCC patients (n=30) in the presence of 50ng/ml IL-4 and 100ng/ml GM-CSF for five days. The cells were cultured for two additional days in the medium and stimulated with 0.1 KE/ml OK432. On day 7, DCs were harvested for injection, 5x106 cells suspended in 5ml normal saline containing 1% autologous plasma, and injected into HCC with a needle percutaneously after RFA. Adverse events were monitored clinically and biochemically after DC infusion. RESULTS: DCs [HLA-DR(+)CD86(+)CD14(-)] derived from HCC patients contained large proportions of myeloid subsets [CD11 c(+)CD123(-)] when analyzed by flow cytometry. OK432 stimulation developed DCs expressing high levels of CD80, 83, 86 and CCR7, producing Th1-type cytokines (IL-12 and IFNγ) quantitated by Bioplex assay and displaying high stimulatory capacity in allo-genic mixed leukocyte reaction (MLR) (P < 0.01) compared to immature DC. There were no grades III or IV National Cancer Institute Common Toxicity Criteria adverse events and also no clinical or serological evidence of hepatic failure or autoimmune response in any patients. Kaplan-Meier analysis indicated that patients treated with RFA and OK432-stimulated DC transfer had tended to prolonged recurrence-free survival (360 days after the treatment) compared with historical controls that had been treated with RFA alone. CONCLUSIONS: Concurrent treatment with OK432-stimulated DC infusion can be performed safely at the same time as RFA in patients with HCC. This study suggests that its usage with locoregional treatments may enhance anti-tumor response against HCC.


Shuichi Kaneko - Grant/Research Support: MDS, Co., Inc, Chugai Pharma., Co., Inc, Toray Co., Inc, Daiichi Sankyo., Co., Inc, Dainippon Sumitomo, Co., Inc, Aji-nomoto Co., Inc, MDS, Co., Inc, Chugai Pharma., Co., Inc, Toray Co., Inc, Daiichi Sankyo., Co., Inc, Dainippon Sumitomo, Co., Inc, Ajinomoto Co., Inc, Bayer Japan

The following people have nothing to disclose: Masaaki Kitahara, Eishiro Mizukoshi, Kiichiro Kaji, Kazutoshi Yamada, Hidetoshi Nakagawa, Hajime Sunagozaka, Kuniaki Arai, Tatsuya Yamashita


Computational insights into the role of USP18 in type I interferon refractoriness

Chitra Raju Nayak1, Vera A. Cherepanov4, JordanJ. Feld2,3, Anton Zilman1;

1 Department of Physics, University of Toronto, Toronto, ON, Canada; 2Toronto Western Hospital, Toronto, ON, Canada; 3McLaughlin-Rotman Centre for Global Health, University of Toronto, Toronto, ON, Canada; 4MaRS DISCOVERY TWR-TMDT, Toronto, ON, Canada

Background: Type I interferons are used effectively in the treatment of Hepatitis C by activating a cascade of interferon-stimu-lated genes with antiviral properties. The signalling cascade involves the binding of IFN to the 2 subunits of the IFN receptor, IFNAR1 (R1) and IFNAR2 (R2), to form a ternary complex. The kinases - Jak's and Tyk's - bound to the cytoplasmic domains of receptor subunits become phosphorylated, which further phosphorylates STAT( p-STAT). Dimers of p-STAT migrate to the nucleus to initiate the transcription of a large number of genes. Type I interferons exhibit a reduced response (refractoriness) to prolonged or multiple doses of IFN. It has been shown that despite binding to the same receptor, IFN-α is more refractory than IFN-β and USP18 plays a role in the refractory state. Methods: We have used a mathematical modeling approach to better understand the determinants of the refractory state, which may be key to improving IFN responsiveness in patients treated with IFN-based therapy .The association and dissociation of the IFN's to the receptor subunits and the phosphorylation of STAT is simulated using the Gillespie stochastic simulation algorithm. The three dimensional and two dimensional association and dissociation rates of IFN α and β are informed by published data. The unavailable rates are evaluated from the principle of detailed balance that requires certain relations to be obeyed by the reaction rates in equilibrium/steady state. The results obtained by numerical simulations are verified by analytic solutions. In order to investigate the refractory behavior, we allow Jak or USP18 to bind to the R2 subunit in our model. However only R2 bound to Jak can activate STAT and thus contribute to downstream signalling. Results: Our model reproduced the experimentally observed results that IFN β, which binds strongly to both the subunits and forms more ternary complexes than α, shows less refractoriness.. USP18 binding to R2 caused the number of active complexes formed by IFNα and IFNβ to drop in an identical way. However, the relative abundance of the IFNAR subunits and differing affinities of IFN α and β for the receptor can explain the differential refractoriness of the type I IFNs. Conclusion: Our model predicts that the main determinant of the refractory state is the number of active IFN receptors on treated cells, which in turn depends on the affinity of the IFN for its receptor. Varying the ratio of IFNAR subunits and the affinity of the IFN for the receptor has the potential to make cells less refractory and thus more responsive to IFN treatment. Our results agree well with the experimental data.


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

The following people have nothing to disclose: Chitra Raju Nayak, Vera A. Cherepanov, Anton Zilman


Kinetics of lymphocyte activation and senescence markers in chronic HCV-infected patients on PegIFNα/Rib-avirin/Telaprevir

Celine Cognet1, Juliette Foucher2, Pascale Trimoulet3, Julien Vergniol2, Wassil Merrouche2, Jean francois Moreau1, Jean Luc Taupin1, Linda Wittkop4, Victor de Ledinghen2, Isabelle Pellegrin 1;

1 Immunology, CHU Bordeaux, Bordeaux, France; 2Hepato gas-troenterology, CHU Pessac, Pessac, France; 3Virology, CHU Bordeaux, Bordeaux, France; 4Medical Information Service, CHU Bordeaux, Bordeaux, France

INTRODUCTION: Immune activation described in chronic viral infections (CMV, HIV) or in healthy elderly population is associated with an increase in morbidity and mortality in the context of immune senescence, defining the concept of inflamm-aging. No data was reported on characterization of this immune status in HCV-infected patients with or without HCV replication. Our aim was to describe the immunological markers of activation, exhaustion and senescence in a population of chronic HCV-infected patients initiating a Peg-IFNα/RBV/Telaprevir therapy. METHODS: Patients with a prior PegIFNα/RBV non response who had initiated (D0) a successful PegIFNα/RBV/Telaprevir therapy (undetectable HCV-RNAfrom Week (W) 12 to W72) were included in the study. The immuno logical markers were measured at D0 and W12 and compared to 20 healthy donors (HD): CD4+ and CD8+ activation (DR+), maturation (TN: CD45RA+CD27+, TCM: CD45RA-CD27+, TEM: CD45RA-CD27-, TEMRA: CD45RA+CD27-), senescent subsets (CD57+CD28-), regulatory T cells

(CD4+CD25high,CD127low) and PD1 on DR+LT cells as surrogate marker of exhaustion. RESULTS: 37 patients (M=24) were analyzed: median age=54 years, median duration of infection=32 years; median HCV-RNA=11419000IU/mL; 46% IL28rs8099917TT; transient elastography median value=10kPa. As expected, total and subsets lymphocyte counts dropped on PegIFNα/RBV/Telaprevir therapy. There were no significant differences for the LTCD4+ and LTCD8+ senescence, maturation, CD4/CD8 ratio between patients before treatment and HD. LTCD4+DR+ and LTCD8+DR+ were higher in patients (p=10–3 and 0.06 respectively) than in HD, although all values were within the normal ranges. Between D0 and W12 of tritherapy, a significant decrease in percentages of CD4+ and CD8+ senescent (p=10–4) and memory cells (TEM, p=10–2; TEMRA, p=10–4) was observed with no variation in the CD4+DR+ and CD8+DR+ subsets. Percentages of Treg (p=10–3) and CD4+ DR+PD1 + and CD8+ DR+PD1 + (p=10–3) increased significantly in the same period of time. CONCLUSION: High level of HCV exposure for more than 30 years is associated neither with a detectable immune activation in the peripheral blood compartment nor with significant senescent status. We speculated that the decrease in memory and senescent lymphocyte subsets on successful IFN-based therapy may be due to viral replication suppression and/or to variations in lymphocytes homing. Nonetheless, the intrahepatic immune response does exist and may be under the regulation of the increase in Treg and in PD1 expression on activated T cells. This observed immune paradox may be of interest for the deciphering of new therapeutic strategies.


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

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: Celine Cognet, Pascale Trimoulet, Julien Vergniol, Wassil Merrouche, Jean francois Moreau, Jean Luc Taupin, Linda Wittkop, Isabelle Pellegrin


Modulation of suPAR by chronic liver inflammation

Athina Chounta, Vassiliki Tzanetakou, Christakis G. Ellinas, Fani Pliarchopoulou, Thomas Tsaganos, Antigoni Kotsaki, Virginia Mplani, Evangelos Giamarellos-Bourboulis;

4th Department of Internal Medicine, University of Athens, “ATTIKON” Hospital, Athens, Greece

Objective: Urokinase plasminogen activator receptor (uPAR) is located on neutrophil cell membranes. The soluble form suPAR is increased in chronic infection by the human immunodeficiency virus and it is predictive of outcome. This prompted us to study the kinetics of suPAR in chronic liver inflammation where no data exist. Methods: suPAR was measured by an enzyme immunoassay in the serum of 28 healthy volunteers and of 275 patients with chronic liver inflammation defined as any more than 2-fold increase of serum aminotransferases for more than six months. Results: Median suPAR were (p values refer to comparisons with healthy controls): 3.51 ng/ml for controls; 6.89 ng/ml for 99 patients with chronic hepatitis B (p< 0.0001); 7.57 ng/ml for 103 patients with chronic hepatitis C (p< 0.0001); 4.71 ng/ml for 29 patients with autoimmune hepatitis (p: 0.004); 3.36 ng/ml for 42 patients with non alcoholic fatty liver disease (NAFLD) (p: 0.606); and 6.89 ng/ml for 3 patients with alcoholic steatohepatitis (p< 0.0001). Using quar-tile distribution, 60 patients with stage of fibrosis between 4 and 6 (Ishak) and belonging to the upper quartile of distribution were classified with advanced fibrosis. Median suPAR was 6.39 ng/ml in less advanced fibrosis and 8.51 ng/ml in advanced fibrosis respectively (p< 0.0001). After ROC analysis, suPAR greater than 8.98 ng/ml had 90.6% specificity to indicate patients at advanced fibrosis (odds ratio: 8.50, 95% CI: 4.23–17.07). A positive correlation was found between serum suPAR and the viral load (rs: +0.271, p: 0.008) and the grade of inflammation (rs: +0.384, p< 0.0001) of HBV patients. No respective correlations were found on HCV patients. Conclusions: suPAR is increased in chronic liver inflammation particularly in fibrosis; Although it can be used as an index of advanced fibrosis, kinetics are largely affected in HBV.


The following people have nothing to disclose: Athina Chounta, Vassiliki Tzanetakou, Christakis G. Ellinas, Fani Pliarchopoulou, Thomas Tsaganos, Antigoni Kotsaki, Virginia Mplani, Evangelos Giamarellos-Bourboulis