Ito M, Watanabe M, Kamiya H, Sakurai M
Department of Pediatrics, Mie University School of Medicine, MieJapan.
J Virol Methods. 1996 Jan;56(1):77-84. doi: 10.1016/0166-0934(95)01954-5.
Cell-mediated cytotoxicity against cytomegalovirus (CMV)-infected fibroblasts (FS-4 cells) was investigated by a non-radioactive assay, and by DNA fragmentation ELISA and LDH release assay and the assays were compared to the standard chromium release assay. Fragmentation of DNA and LDH activity were detected in the supernatant of CMV-infected FS-4 cells cultured with non-adherent peripheral blood mononuclear cells (PBMC). The DNA fragmentation ELISA was most sensitive to cytotoxicity against CMV-infected FS-4 cells and showed excellent correlation with the standard chromium release assay. DNA fragmentation of CMV-infected FS-4 cells by non-adherent PBMC was reduced markedly by treatment with anti-leu 11b plus complement. Thus, the present DNA fragmentation ELISA is non-radioactive, highly sensitive and a useful method for detecting natural killer cell-mediated cytotoxicity against CMV-infected fibroblasts.
通过一种非放射性检测方法研究了针对巨细胞病毒（CMV）感染的成纤维细胞（FS-4细胞）的细胞介导细胞毒性，并与DNA片段化ELISA和乳酸脱氢酶（LDH）释放检测进行了比较，且这些检测与标准的铬释放检测进行了对比。在用非贴壁外周血单核细胞（PBMC）培养的CMV感染的FS-4细胞的上清液中检测到了DNA片段化和LDH活性。DNA片段化ELISA对针对CMV感染的FS-4细胞的细胞毒性最为敏感，并且与标准的铬释放检测显示出极好的相关性。用抗Leu 11b加补体处理后，非贴壁PBMC对CMV感染的FS-4细胞的DNA片段化明显减少。因此，目前的DNA片段化ELISA是非放射性的、高度敏感的，并且是一种检测自然杀伤细胞介导的针对CMV感染成纤维细胞的细胞毒性的有用方法。
