Non-radioactive assay of natural killer cell-mediated cytotoxicity against cytomegalovirus-infected fibroblasts by DNA fragmentation ELISA.
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.
Ito
M
M
Department of Pediatrics, Mie University School of Medicine, MieJapan.
Watanabe
M
M
Kamiya
H
H
Sakurai
M
M
D000328:Adult; D002460:Cell Line; D002857:Chromium / Q000378:metabolism; D003587:Cytomegalovirus / Q000276:immunology*; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D003602:Cytotoxicity, Immunologic; D004247:DNA / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005347:Fibroblasts / Q000276:immunology / Q000821:virology*; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology*
D002857:Chromium; D004247:DNA