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Lymphotropic HCV strain can infect human primary naïve CD4+ cells and affect their proliferation and IFN-γ secretion activity

  • Original Article—Liver, Pancreas, and Biliary Tract
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Abstract

Background

Lymphotropic hepatitis C virus (HCV) infection of B and T cells might play an important role in the pathogenesis of hepatitis C. Recently, we showed that a lymphotropic HCV (SB strain) could infect established T-cell lines and B-cell lines. However, whether HCV replication interferes with cell proliferation and function in primary T lymphocytes is still unclear.

Aim

The aim of this study was to analyze whether HCV replication in primary T lymphocytes affected their development, proliferation, and Th1 commitment.

Methods

SB strain cell culture supernatant (2 × 104 copies/ml HCV) was used to infect several kinds of primary lymphocyte subsets. Mock, UV-irradiated SB-HCV, JFH-1 strain, and JFH-1 NS5B mutant, which could not replicate in T cells, were included as negative controls. Carboxyfluorescein succinimidyl ester (CFSE) and CD45RA double staining was used to evaluate the proliferative activity of CD4+CD45RA+CD45RO− naïve CD4+ cells. Interferon (IFN)-γ and interleukin (IL)-10 secretion assays magnetic cell sorting (MACS) were carried out.

Results

Negative strand HCV RNA was detected in CD4+, CD14+, and CD19+ cells. Among CD4+ cells, CD4+CD45RA+RO− cells (naïve CD4+ cells) were most susceptible to replication of the SB strain. The levels of CFSE and CD45RA expression gradually declined during cell division in uninfected cells, while HCV-infected naïve CD4+ cells expressed higher levels of CFSE and CD45RA than Mock or UV-SB infected naïve CD4+ cells. Moreover, the production of IFN-γ was significantly suppressed in SB-infected naïve CD4+ cells.

Conclusions

Lymphotropic HCV replication suppressed proliferation and development, including that towards Th1 commitment, in human primary naïve CD4+ cells.

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Acknowledgments

This work was supported in part by a Grant-in-Aid from the Ministry of Education, Culture, Sport, Science, and Technology of Japan (#21790642 to Y.K.), and by Health and Labour Sciences Research Grants for Research on Measures for Intractable Diseases (from the Ministry of Health, Labour and Welfare of Japan (to Y.U.). We are grateful to Dr. Michael M.C. Lai for providing the SB-HCV strain, and to Dr. Takaji Wakita for providing pJFH-1 and pJFH-1/GND.

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Authors and Affiliations

  1. Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aobaku, Sendai, 980-8574, Japan

    Yasuteru Kondo, Yoshiyuki Ueno, Eiji Kakazu, Masaaki Shiina, Keiichi Tamai, Jun Inoue, Yuta Wakui, Koji Fukushima, Noriyuki Obara, Osamu Kimura & Tooru Shimosegawa

  2. Department of Molecular Microbiology and Immunology, University of Southern California Keck School of Medicine, 2011 Zonal Avenue, Los Angeles, CA, 90033, USA

    Keigo Machida

  3. School of Health Science, Tohoku University, 1-1 Seiryo, Aobaku, Sendai, 980-8574, Japan

    Koju Kobayashi

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  1. Yasuteru Kondo
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  2. Yoshiyuki Ueno
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Corresponding author

Correspondence to Yoshiyuki Ueno.

Electronic supplementary material

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535_2010_297_MOESM1_ESM.ppt

Suppl. Fig. 1 HCV infection of primary naïve CD4+ cells. Representative gels shows the semiquantitative detection of negative strand HCV-RNA in CD45RA+RO− naïve CD4+ cells 2 days and 7 days after infection. (PPT 128 kb)

535_2010_297_MOESM2_ESM.ppt

Suppl. Fig. 2 Infectivity of SB-HCV in the naïve CD4+ cells. The percentage of NS5A expressing cells was determined by counting the numbers of positive stained cells in 400 cells. As a negative control, cells treated with UV-irradiated HCV were used. (PPT 1069 kb)

535_2010_297_MOESM3_ESM.ppt

Suppl. Fig. 3 The expression levels of HCV individual proteins and transfection efficiencies. The expression levels of HCV individual proteins were shown in these histograms. The numbers in these histograms indicate the percentage of proteins-positive cells. (PPT 488 kb)

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Kondo, Y., Ueno, Y., Kakazu, E. et al. Lymphotropic HCV strain can infect human primary naïve CD4+ cells and affect their proliferation and IFN-γ secretion activity. J Gastroenterol 46, 232–241 (2011). https://doi.org/10.1007/s00535-010-0297-2

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  • DOI: https://doi.org/10.1007/s00535-010-0297-2

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