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


OBJECTIVE
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 × 10(4) 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.
Kondo
Yasuteru
Y
Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aobaku, Sendai 980-8574, Japan.
Ueno
Yoshiyuki
Y
Kakazu
Eiji
E
Kobayashi
Koju
K
Shiina
Masaaki
M
Tamai
Keiichi
K
Machida
Keigo
K
Inoue
Jun
J
Wakui
Yuta
Y
Fukushima
Koji
K
Obara
Noriyuki
N
Kimura
Osamu
O
Shimosegawa
Tooru
T
D017209:Apoptosis; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D002455:Cell Division; D049109:Cell Proliferation; D002478:Cells, Cultured; D005452:Fluoresceins / Q000378:metabolism; D016174:Hepacivirus / Q000254:growth & development*; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism*; D016753:Interleukin-10 / Q000378:metabolism; D017493:Leukocyte Common Antigens / Q000378:metabolism; D013388:Succinimides / Q000378:metabolism; D014162:Transfection; D014758:Viral Core Proteins / Q000378:metabolism
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D013388:Succinimides; D014758:Viral Core Proteins; D016753:Interleukin-10; D007371:Interferon-gamma; D017493:Leukocyte Common Antigens