Posavad C M, Rosenthal K L
Department of Pathology, McMaster University Health Sciences Centre, Hamilton, Ontario, Canada.
J Virol. 1992 Nov;66(11):6264-72. doi: 10.1128/JVI.66.11.6264-6272.1992.
We examined the ability of human anti-herpes simplex virus (HSV) cytotoxic T lymphocytes (CTL) to lyse autologous human fibroblasts infected with HSV. In contrast to HSV-infected human Epstein-Barr virus-transformed B cells (LCL), which were lysed by HLA-restricted anti-HSV CTL, autologous fibroblasts infected with HSV were resistant to lysis. This resistance was not due to a lack of infectivity or production of HSV proteins since greater than 90% of the cells were infected and expressed abundant levels of viral proteins. HSV-infected human fibroblasts were also tested for susceptibility to lysis by alloantigen-specific CTL. Although allogeneic LCL and uninfected allogeneic fibroblasts were killed, human fibroblasts infected with HSV demonstrated a time-dependent resistance to lysis by alloantigen-specific CTL. HSV-infected human fibroblasts were not resistant to all forms of cell-mediated cytotoxicity since they were sensitive to antibody-dependent cellular cytotoxicity. Although one may suspect that the resistance of HSV-infected human fibroblasts to anti-HSV CTL and alloantigen-specific CTL-mediated lysis was due to a lack of major histocompatibility complex expression, Confer et al. (Proc. Natl. Acad. Sci. USA 87:3609-3613, 1990) previously demonstrated that incubation of human natural killer and lymphokine-activated killer cells with monolayers of human fibroblasts infected with HSV "disarmed" the killers in that they were unable to lyse sensitive target cells. We extend their results and show that incubation of anti-HSV CTL or alloantigen-specific CTL with uninfected fibroblasts did not affect their lytic activity, whereas CTL incubated with HSV-infected fibroblasts for 2 to 6 h rendered the CTL incapable of lysing their normally sensitive target cells. Indeed, human fibroblasts infected for merely 2 h with HSV were able to profoundly inhibit the cytotoxic activity of alloantigen-specific CTL. Thus, HSV-infected human fibroblasts are not inherently resistant to lysis by anti-HSV CTL or alloantigen-specific CTL, but rather contact of CTL with HSV-infected fibroblasts resulted in inactivation of the CTL. The inactivation of CTL appears to be HSV specific since incubation of alloantigen-specific CTL in sandwich assays with fibroblasts infected with HSV type 1 (HSV-1) or HSV-2 resulted in inactivation, whereas incubation of CTL with fibroblasts infected with adenovirus or vaccinia virus had no effect. Further, although incubation of alloantigen-specific CTL in sandwich assays with HSV-infected fibroblasts resulted in inhibition of CTL activity, exposure of CTL in Transwell cultures to cell-free supernatant from HSV-infected fibroblasts did not mediate this inhibitory effect.(ABSTRACT TRUNCATED AT 400 WORDS)
我们检测了人类抗单纯疱疹病毒（HSV）细胞毒性T淋巴细胞（CTL）对感染HSV的自体人成纤维细胞进行裂解的能力。与被HLA限制的抗HSV CTL裂解的感染HSV的人类EB病毒转化B细胞（LCL）不同，感染HSV的自体成纤维细胞对裂解具有抗性。这种抗性并非由于缺乏感染性或HSV蛋白的产生，因为超过90%的细胞被感染并表达了大量的病毒蛋白。我们还检测了感染HSV的人类成纤维细胞对同种异体抗原特异性CTL裂解的敏感性。虽然同种异体LCL和未感染的同种异体成纤维细胞被杀死，但感染HSV的人类成纤维细胞对同种异体抗原特异性CTL的裂解表现出时间依赖性抗性。感染HSV的人类成纤维细胞对所有形式的细胞介导细胞毒性并不具有抗性，因为它们对抗体依赖性细胞毒性敏感。尽管有人可能怀疑感染HSV的人类成纤维细胞对抗HSV CTL和同种异体抗原特异性CTL介导的裂解具有抗性归因于主要组织相容性复合体表达的缺乏，但Confer等人（Proc. Natl Acad. Sci. USA 87:3609 - 361 3，1990）先前证明，将天然杀伤细胞和淋巴因子激活的杀伤细胞与人感染HSV的成纤维细胞单层孵育会使杀伤细胞 “失活”，因为它们无法裂解敏感靶细胞。我们在此结果之上进一步发现抗HSCTL或同种异体抗原特异性CTL与未感染的成纤维细胞孵育不会影响其裂解活性，而与感染HSV的成纤维细胞孵育2至6小时会使CTL无法裂解其正常敏感靶细胞。事实上仅感染HSV 2小时的人类成纤维细胞就能显著抑制同种异体抗原特异性CTL的细胞毒性活性。因此，感染HSV的人类成纤维细胞并非固有地对抗HS CTL或同种异体抗原特异性CTL介导的裂解具有抗性，而是CTL与感染HSV的成纤维细胞接触导致CTL失活。CTL的失活似乎具有HSV特异性，因为在夹心试验中，将同种异体抗原特异性CTL与感染1型HSV（HS‑1）或HSV‑2的成纤维细胞孵育会导致失活，而将CTL与感染腺病毒或痘苗病毒的成纤维细胞孵育则没有影响。此外，虽然在夹心试验中将同种异体抗原特异性CTL与感染HSV的成纤维细胞孵育会导致CTL活性受到抑制，但在Transwell培养中，将CTL暴露于感染HSV的成纤维细胞的无细胞上清液中并不会介导这种抑制作用。（摘要截短至400字）
