Wedemeyer H, Mizukoshi E, Davis A R, Bennink J R, Rehermann B
Liver Diseases Section, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20982, USA.
J Virol. 2001 Dec;75(23):11392-400. doi: 10.1128/JVI.75.23.11392-11400.2001.
The cellular immune response contributes to viral clearance as well as to liver injury in acute and chronic hepatitis C virus (HCV) infection. An immunodominant determinant frequently recognized by liver-infiltrating and circulating CD8(+) T cells of HCV-infected patients is the HCV(NS3-1073) peptide CVNGVCWTV. Using a sensitive in vitro technique with HCV peptides and multiple cytokines, we were able to expand cytotoxic T cells specific for this determinant not only from the blood of 11 of 20 HCV-infected patients (55%) but also from the blood of 9 of 15 HCV-negative blood donors (60%), while a second HCV NS3 determinant was recognized only by HCV-infected patients and not by seronegative controls. The T-cell response of these healthy blood donors was mediated by memory T cells, which cross-reacted with a novel T-cell determinant of the A/PR/8/34 influenza A virus (IV) that is endogenously processed from the neuraminidase (NA) protein. Both the HCV NS3 and the IV NA peptide displayed a high degree of sequence homology, bound to the HLA-A2 molecule with high affinity, and were recognized by cytotoxic T lymphocytes with similar affinity (10(-8) M). Using the HLA-A2-transgenic mouse model, we then demonstrated directly that HCV-specific T cells could be induced in vivo by IV infection. Splenocytes harvested from IV-infected mice at the peak of the primary response (day 7 effector cells) or following complete recovery (day 21 memory cells) recognized the HCV NS3 peptide, lysed peptide-pulsed target cells, and produced gamma interferon. These results exemplify that host responses to an infectious agent are influenced by cross-reactive memory cells induced by past exposure to heterologous viruses, which could have important consequences for vaccine development.
细胞免疫反应在急性和慢性丙型肝炎病毒(HCV)感染中有助于病毒清除以及肝损伤。HCV感染患者肝脏浸润和循环CD8(+) T细胞频繁识别的一个免疫显性决定簇是HCV(NS3 - 1073)肽CVNGVCWTV。我们使用一种针对HCV肽和多种细胞因子的灵敏体外技术,不仅能够从20例HCV感染患者中的11例(55%)血液中扩增出针对该决定簇的细胞毒性T细胞,还能从15例HCV阴性献血者中的9例(60%)血液中扩增出此类细胞毒性T细胞,而第二个HCV NS3决定簇仅被HCV感染患者识别,血清阴性对照则不能识别。这些健康献血者的T细胞反应由记忆T细胞介导,这些记忆T细胞与从神经氨酸酶(NA)蛋白内源性加工而来的A/PR/8/34甲型流感病毒(IV)的一个新T细胞决定簇发生交叉反应。HCV NS3肽和IV NA肽均显示出高度的序列同源性,以高亲和力结合到HLA - A2分子上,并被细胞毒性T淋巴细胞以相似的亲和力(10(-8) M)识别。然后,我们使用HLA - A2转基因小鼠模型直接证明,IV感染可在体内诱导产生HCV特异性T细胞。在初次反应高峰期(第7天效应细胞)或完全恢复后(第21天记忆细胞)从IV感染小鼠收获的脾细胞识别HCV NS3肽,裂解肽脉冲靶细胞,并产生γ干扰素。这些结果表明,宿主对一种感染因子的反应受到既往接触异源病毒诱导产生的交叉反应性记忆细胞的影响,这可能对疫苗开发产生重要影响。