对鼠巨细胞病毒的细胞毒性反应：产生MCMV特异性靶细胞的条件。

The cytotoxic response to murine cytomegalovirus: requirements for the generation of MCMV-specific target cells.

作者信息

Sinickas V G, Ashman R B, Blanden R V

出版信息

Immunol Cell Biol. 1987 Apr;65 ( Pt 2):173-82. doi: 10.1038/icb.1987.19.

DOI:10.1038/icb.1987.19

Abstract

The preparation of target cells susceptible to lysis by murine cytomegalovirus (MCMV)-immune T cells in vitro was investigated and found to be dependent upon target cell type, culture conditions, virus adsorption protocol and virus preparation. Optimally sensitive MCMV-infected targets were obtained by preculture of mouse embryo fibroblasts (MEF) in 3% vol/vol concanavalin-A-stimulated spleen cell supernatant (CSS)-supplemented medium, adsorption of salivary gland stock MCMV under 800 g centrifugation and at least 4 h further incubation at 37 degrees before addition of T cells. In contrast, salivary gland stock MCMV did not cause thioglycollate-induced peritoneal exudate cells to be susceptible to MCMV-specific T cell-mediated lysis, whereas tissue culture-passaged stock MCMV was successful. The preparation of MCMV-infected target cells is discussed in terms of the need for H-2 and viral antigen expression for T cell recognition.

摘要

对体外培养的易被鼠巨细胞病毒（MCMV）免疫T细胞裂解的靶细胞制备方法进行了研究，发现其取决于靶细胞类型、培养条件、病毒吸附方案和病毒制剂。通过在含3%（体积/体积）伴刀豆球蛋白A刺激的脾细胞上清液（CSS）的培养基中预培养小鼠胚胎成纤维细胞（MEF），在800g离心条件下吸附唾液腺来源的MCMV原种，并在37℃下进一步孵育至少4小时后再加入T细胞，可获得对MCMV感染最敏感的靶细胞。相比之下，唾液腺来源的MCMV原种不会使巯基乙酸盐诱导的腹腔渗出细胞易被MCMV特异性T细胞介导的裂解，而组织培养传代的原种MCMV则成功诱导了裂解。本文根据T细胞识别所需的H-2和病毒抗原表达情况，对MCMV感染靶细胞的制备进行了讨论。