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Serum amyloid A has antiviral activity against hepatitis C virus by inhibiting virus entry in a cell culture system†
Article first published online: 28 NOV 2006
Copyright © 2006 American Association for the Study of Liver Diseases
Volume 44, Issue 6, pages 1626–1634, December 2006
How to Cite
Lavie, M., Voisset, C., Vu-Dac, N., Zurawski, V., Duverlie, G., Wychowski, C. and Dubuisson, J. (2006), Serum amyloid A has antiviral activity against hepatitis C virus by inhibiting virus entry in a cell culture system. Hepatology, 44: 1626–1634. doi: 10.1002/hep.21406
Potential conflict of interest: Nothing to report.
- Issue published online: 28 NOV 2006
- Article first published online: 28 NOV 2006
- Manuscript Accepted: 6 SEP 2006
- Manuscript Received: 14 MAR 2006
- Agence Nationale de Recherche sur le SIDA et les Hépatites Virales (ANRS) and ARC
- Howard Hughes Medical Institute
Supplementary material for this article can be found on the H EPATOLOGY website ( http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html ).
|jws-hep.21406.fig1.pdf||31K||Supplementary Fig. 1. ApoA-I and ApoE do not affect HCVpp and HCVcc infectivities.(A) ApoA-I and ApoE do not affect HCVpp infectivity. Huh-7 cells were infected with HCVpp in presence or absence of ApoA-I or ApoE (50 μg/mL in LPDS medium). After virus adsorption, cells were washed and further incubated for 2 days. Cells were then lyzed and processed to measure the luciferase activities. The results are presented as percentages of infectivity of pseudotyped particles incubated in the presence of ApoA-I or ApoE relative to infectivity of untreated pseudotyped particles and represent the average ? standard deviation of three independent experiments. (B and C) ApoA-I and ApoE do not affect HCVcc infectivity. Huh-7 cells were infected with HCVcc in the presence of ApoA-I or ApoE during the 2-hour period of virus adsorption. At 2 days post-infection, cells were lyzed and HCVcc infectivity was analyzed as in Fig. 1.|
|jws-hep.21406.fig2.pdf||37K||Supplementary Fig. 2. The antiviral activity of SAA is not due to its action on target cells.(A) Preincubation of cells with SAA at 37°C does not affect HCVcc infectivity. Huh-7 cells were preincubated at 37°C in the presence or absence of SAA (15 μg/mL). After 3 hours of incubation, cells were washed and incubated with HCVcc for 30 min. In a parallel experiment, Huh-7 cells were incubated with HCVcc for 30 minutes in the presence of SAA. After virus adsorption, cells were washed and further incubated at 37°C. At 2 days post-infection, cells were lysed and HCVcc infectivity was analyzed as in Fig. 1. (B) SAA does not compete with HCVcc for binding to Huh-7 cells at 4°C. Huh-7 cells were preincubated for 90 min at 4°C in LPDS medium with or without SAA (15 μg/mL). Cells were then washed and incubated for 90 min at 4°C with HCVcc in the presence or absence of SAA (15 μg/mL). After virus adsorption at 4°C, cells were washed and further incubated at 37°C. At 2 days post-infection, cells were lyzed and HCVcc infectivity was analyzed as in Fig. 1.|
|jws-hep.21406.fig3.pdf||21K||Supplementary Fig. 3. SAA does not affect the binding of soluble E2 on CHO-SR-BI cells.The E2/SR-BI binding assay has been described by Scarselli et al. 2002 (8). Briefly, CHO cells were transfected with phCMV-SR-BI (kindly provided by B. Bartosch, INSERM U758, Lyon, France). After 48 hours of incubation, cells were detached with PBS 2 mM EDTA and washed twice with PBS before being incubated either with E2 or control concentrated supernatant or E2 preincubated with SAA 0.65 mg/mL for 2 hours at 37°C. An additional control was made by incubating E2 with CHO cells transfected with an empty vector. After 3 hours of incubation at room temperature, cells were washed and incubated with dithiobis-sulfosuccinimidyl-propionate (DTSSP, Pierce) crosslinker at 2 mM in PBS for 30 min at room temperature. The reaction was stopped by addition of 50 mM Tris-HCl pH 7.5. Cells were lysed in PBS 1% Triton X-100 in the presence of protease inhibitors. E2-receptor complexes were then immunoprecipitated with Mab H53 (anti-E2) and revealed by western blotting with anti-E2 (3/11) and anti-SR-BI (ClaI, BD Bioscience) antibodies.|
|jws-hep.21406.tbl1.pdf||13K||Supplementary Table 1. SAA is detected in some sera of HCV infected patients (n=94).|
|jws-hep.21406.tbl2.pdf||9K||Supplementary Table 2. SAA is detected in some sera of HBV infected patients (n=28).|
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