Sugamata M, Ewalt L C, Perry L L, Lodmell D L
National Institutes of Health, National Institute of Allergy and Infectious Diseases, Rocky Mountain Laboratories, Hamilton, Montana 59840.
J Virol Methods. 1990 Jul;29(1):1-11. doi: 10.1016/0166-0934(90)90002-w.
Cells persistently infected with Evelyn-Rokitnicki-Abelseth (ERA) rabies virus were established. The cells were used as stimulator and target cells to compare H-2 restricted cytotoxic T lymphocyte (CTL) responses specific for rabies virus in A/WySnJ (H-2a), C57BL/6J (H-2b), BALB/cByJ (H-2d), A.SW/SnJ (H-2s) and SJL/J (H-2s) mice. Using a 51chromium release assay, it was determined that an effector/target (E/T) ratio of 5:1 was necessary to demonstrate specific lysis of ERA virus persistently infected mouse neuroblastoma (MNB) (H-2a), EL-4 (H-2b) and P815 (H-2d) cells. Effectors at an E:T ratio of only 0.05:1 specifically lysed an SV-40 transformed SJL/J mouse fibroblast (SSSV) (H-2s) target monolayer. The CTL destruction of the SSSV monolayer was observed visually following Giemsa staining. This is the first instance in which a detailed method for detection of murine anti-rabies virus CTLs has been reported. Furthermore, it is the first time target cells persistently infected with rabies virus were used as stimulator cells to amplify CTLs in vitro and as target cells in the CTL assay. It also is the initial report in which rabies specific CTLs were characterized in H-2d and H-2s rabies virus infected mice.
建立了持续感染伊夫林 - 罗基特尼基 - 阿贝尔塞思（ERA）狂犬病病毒的细胞。这些细胞用作刺激细胞和靶细胞，以比较A/WySnJ（H-2a）、C57BL/6J（H-2b）、BALB/cByJ（H-2d）、A.SW/SnJ（H-2s）和SJL/J（H-2s）小鼠中针对狂犬病病毒的H-2限制性细胞毒性T淋巴细胞（CTL）反应。使用51铬释放试验确定，效应细胞与靶细胞（E/T）比例为5:1对于证明ERA病毒持续感染的小鼠神经母细胞瘤（MNB）（H-2a）、EL-4（H-2b）和P815（H-2d）细胞的特异性裂解是必要的。仅以0.05:1的E:T比例的效应细胞能特异性裂解SV-40转化的SJL/J小鼠成纤维细胞（SSSV）（H-2s）靶单层。吉姆萨染色后通过肉眼观察到SSSV单层的CTL破坏。这是首次报道检测小鼠抗狂犬病病毒CTL的详细方法。此外，这也是首次将持续感染狂犬病病毒的靶细胞用作刺激细胞在体外扩增CTL，并在CTL试验中用作靶细胞。这也是首次在H-2d和H-2s狂犬病病毒感染的小鼠中鉴定狂犬病特异性CTL的报告。
