Standardization of a micro-cytotoxicity assay for human natural killer cell lytic activity.
Cytotoxicity assays are widely used to evaluate the functional activity of NK and T cells against tumour target cells and the release of radioactive sodium chromate from labelled target cells is still the most commonly used marker of target lysis in culture supernatants. We describe here the standardization of a micro-cytotoxicity test in which the number of cytolytic effector and tumour target cells have been decreased by a factor of 10. The release obtained by 500 tumour target cells was compared with the release obtained by 5000 target cells in the standard cytotoxicity assay for target:effector cell ratios from 1:1 to 1:100. Both gamma and beta emissions of the 51Cr isotope were evaluated to determine the assay release. The results obtained by the micro-cytotoxicity assay (500 target cells) were comparable to those of the standard assay (5000 target cells) and 51Cr release evaluation using the gamma counter was the most sensitive method of determining lytic activity using 500 tumour target cells. beta counter evaluation using solid phase scintillation was found to be a reproducible alternative method, even if the lytic curves cannot be compared with those obtained using the traditional method.
Laboratorio di Immunologia e Genetica, Istituto di Ricerca Codivilla Putti I.O.R., Bologna, Italy.
E
E
Mariani
M C
MC
Monaco
S
S
Sgobbi
J F
JF
de Zwart
A R
AR
Mariani
A
A
Facchini
D002857:Chromium / Q000032:analysis; D002860:Chromium Radioisotopes; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods* / Q000592:standards; D004305:Dose-Response Relationship, Drug; D006801:Humans; D016219:Immunotherapy, Adoptive; D007694:Killer Cells, Natural / Q000276:immunology*; D007951:Leukemia, Myeloid / Q000378:metabolism / Q000628:therapy; D012588:Scintillation Counting; D014407:Tumor Cells, Cultured / Q000528:radiation effects
D002860:Chromium Radioisotopes; D002857:Chromium