Witmer L A, Rosenthal K L, Graham F L, Friedman H M, Yee A, Johnson D C
Department of Pathology, McMaster University, Hamilton, Ontario, Canada.
J Gen Virol. 1990 Feb;71 ( Pt 2):387-96. doi: 10.1099/0022-1317-71-2-387.
In previous work, we observed that H-2k-restricted herpes simplex virus (HSV)-specific cytotoxic T lymphocytes (CTLs) were effectively able to lyse transfected target cells expressing HSV glycoprotein C (gC), but not cells expressing gB, gD or gE. To confirm and extend our observations on the specificity of anti-HSV CTLs, recombinant adenovirus (Ad) vectors able to express HSV-1 gB or gC (AdgB2 or AdgC) were constructed. Syngeneic target cells infected with AdgB2 were efficiently lysed by primary H-2b and H-2d, but not by H-2k-restricted HSV-specific CTL. Limiting dilution studies indicated that 4 to 10% of H-2b-restricted HSV-specific CTLs recognize gB. H-2k, H-2b and H-2d-restricted anti-HSV-1 CTLs were unable to lyse AdgC-infected syngeneic target cells. To examine the apparent discrepancy between the previous results involving transfected H-2k cells expressing gC and the present results involving AdgC-infected cells, gC-expressing cell lines used in previous experiments were subcloned and retested in CTL assays. DC2 cells which were lysed by HSV-specific CTLs in the previous experiments remained sensitive to anti-HSV CTLs but two other clones derived from the same transfection were not lysed. Further, L cells transfected with the gC or gD gene coupled to the mouse mammary tumour virus promoter and capable of expressing high levels of the glycoproteins following dexamethasone induction were not lysed by H-2k-restricted anti-HSV CTLs. These results suggest that HSV-specific CTLs do not recognize gC, at least when it is expressed using an Ad vector and in most transfected cell lines, whereas a significant proportion of anti-viral CTLs recognize gB presented in some but not all murine haplotypes.
在之前的研究中，我们观察到受H-2k限制的单纯疱疹病毒（HSV）特异性细胞毒性T淋巴细胞（CTL）能够有效地裂解表达HSV糖蛋白C（gC）的转染靶细胞，但不能裂解表达gB、gD或gE的细胞。为了证实并扩展我们对抗HSV CTL特异性的观察，构建了能够表达HSV-1 gB或gC的重组腺病毒（Ad）载体（AdgB2或AdgC）。感染AdgB2的同基因靶细胞被原发性H-2b和H-2d高效裂解，但未被受H-2k限制的HSV特异性CTL裂解。有限稀释研究表明，4%至10%的受H-2b限制的HSV特异性CTL识别gB。受H-2k、H-2b和H-2d限制的抗HSV-1 CTL无法裂解感染AdgC的同基因靶细胞。为了研究之前涉及表达gC的转染H-2k细胞的结果与目前涉及感染AdgC的细胞的结果之间明显的差异，对之前实验中使用的表达gC的细胞系进行亚克隆，并在CTL试验中重新测试。在之前的实验中被HSV特异性CTL裂解的DC2细胞对抗HSV CTL仍然敏感，但来自同一转染的另外两个克隆未被裂解。此外，用与小鼠乳腺肿瘤病毒启动子偶联的gC或gD基因转染并在地塞米松诱导后能够表达高水平糖蛋白的L细胞未被受H-2k限制的抗HSV CTL裂解。这些结果表明，HSV特异性CTL不识别gC，至少当它使用Ad载体表达时以及在大多数转染细胞系中是这样，而相当一部分抗病毒CTL识别gB，在一些但不是所有小鼠单倍型中呈递。
