Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo, Aobaku, Sendai 980-8574, Japan.
J Gastroenterol. 2011 Feb;46(2):232-41. doi: 10.1007/s00535-010-0297-2. Epub 2010 Aug 17.
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.
The aim of this study was to analyze whether HCV replication in primary T lymphocytes affected their development, proliferation, and Th1 commitment.
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.
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.
Lymphotropic HCV replication suppressed proliferation and development, including that towards Th1 commitment, in human primary naïve CD4(+) cells.
B 细胞和 T 细胞的淋巴亲嗜性丙型肝炎病毒（HCV）感染可能在丙型肝炎的发病机制中发挥重要作用。最近，我们发现一种淋巴亲嗜性 HCV（SB 株）可以感染已建立的 T 细胞系和 B 细胞系。然而，HCV 复制是否会干扰原代 T 淋巴细胞的增殖和功能仍不清楚。
本研究旨在分析 HCV 在原代 T 淋巴细胞中的复制是否会影响其发育、增殖和 Th1 分化。
使用 SB 株细胞培养上清液（2×104 拷贝/ml HCV）感染几种原代淋巴细胞亚群。假型、紫外线照射 SB-HCV、不能在 T 细胞中复制的 JFH-1 株和 JFH-1 NS5B 突变株被用作阴性对照。使用羧基荧光素琥珀酰亚胺酯（CFSE）和 CD45RA 双重染色来评估 CD4+CD45RA+CD45RO-幼稚 CD4+细胞的增殖活性。进行干扰素（IFN）-γ和白细胞介素（IL）-10 分泌测定和磁细胞分选（MACS）。
在 CD4+、CD14+和 CD19+细胞中检测到负链 HCV RNA。在 CD4+细胞中，CD4+CD45RA+RO-细胞（幼稚 CD4+细胞）最易被 SB 株复制。未感染细胞中，细胞分裂过程中 CFSE 和 CD45RA 的表达逐渐下降，而 HCV 感染的幼稚 CD4+细胞表达的 CFSE 和 CD45RA 高于假型或紫外线照射 SB 感染的幼稚 CD4+细胞。此外，SB 感染的幼稚 CD4+细胞 IFN-γ的产生明显受到抑制。
淋巴亲嗜性 HCV 复制抑制了人类原代幼稚 CD4+细胞的增殖和发育，包括向 Th1 分化的过程。
