Open Access Research

Characterization of antigenic variants of hepatitis C virus in immune evasion

Jane H Wang123*, Matthew J Pianko2, Xiaogang Ke1, Alex Herskovic2, Ronald Hershow4, Scott J Cotler1, Weijin Chen5, Zheng W Chen3 and Lijun Rong3*

Author Affiliations

1 Section of Hepatology, Department of Medicine, University of Illinois at Chicago, Illinois, USA

2 Division of Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, the Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA

3 Department of Microbiology and Immunology, University of Illinois at Chicago, Illinois, USA

4 Division of Epidemiology and Biostatistics, School of Public Health, University of Illinois at Chicago, Illinois, USA

5 Changchun Institute of Biological Products, China National Biotec Group Int. Changchun, China

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Virology Journal 2011, 8:377  doi:10.1186/1743-422X-8-377

Published: 29 July 2011

Additional files

Additional file 1:

Table S1. The information of HCV-infected patients. The information of HCV-infected patients.

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Additional file 2:

Figure S1. An example of CD4+CD25+ cells achieved by culturing HCV-infected PBMCs with variant peptide pool VP1 at presence (B) or absence (A) of anti-TGF-βantibodies; and CD4+CD25+ cells achieved by culturing HCV-infected PBMCs with (D) or without (C) TGF-β+cell deletion. Either anti-TGF-β antibodies or deletion of TGF-β+cells significantly reduced CD4+CD25+ cells. Two representatives of four experiments are shown.

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Figure S2. CD4+CD25+ and CD4+CD25- Tregs of a HCV- healthy individual. Experiments, controls, and data analysis were the same as described in Figure 4. A: Percentages of CD25+ and CD25- gated CD4+ cells. B: Percentages of CD25+ Th3 and Tr1 cells. C: Percentages of CD25- Th3 and Tr1 cells. A single representative of three experiments is shown.

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Figure S3. Treg phenotypes induced by peptide NS3358-375 and variant pools VP1 and VP2 at about 1.5 years post HCV infection. Cell culture and purification of CD4+ cells and controls were the same as described in Figure 4. Intracellular staining of purified CD4+ cells was performed using fluorescent-labeled antibodies to human CD25, IFN-γ, TGF-β and IL-10. A: Percentages of CD25-IL-10+ Tr1 cells. B: Percentages of CD25+IL-10+ Tr1 cells. A single representative of three experiments is shown.

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Figure S4. Treg phenotypes from a patient infected with HCV for over 20 years. Cell culture, purification of CD4+ cells, controls, intracellular staining and flow cytometry analysis were the same as described in Figure 2. Note that VP1 peptides induced higher CD25-Il-10+ Tr1 cells than wild type peptide NS3358-375, VP2 peptides and medium alone.

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Additional file 6:

Figure S5. Serum levels of IL-10 and TGF-β. C: HCV- healthy controls; P: HCV+ patients.

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Additional file 7:

Figure S6. HCV variants induced incomplete T cell activation and/or up-regulated CD45RBlow Tregs. PBMCs were cultured with peptide NS3358-375 (5 μM) and variant pools (1 μM/each peptide) for 66 hours followed by positive selection of CD4+ T cells with anti-human CD4 antibodies and magnetic beads. The CD4+ cells obtained were then stained with fluorescent-labeled antibodies to human CD134 (OX40), and CD45RB and analyzed using a three-color flow cytometer. PBMCs cultured with medium alone were used as control. The effect of peptide NS3358-375 and variant pools VP1 and VP2 on expression of CD134 (A) and CD45RB (B) on CD4 T cells is demonstrated. A representative of two experiments is shown.

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