A non-radioisotopic human natural killer cell assay using rhodamine-123 fluorescent dye as labelling probe

A non-radioisotopic method for assessment of human natural killer (NK) cell activity in peripheral blood mononuclear cells (PBMC) was established by labelling K562 erythroleukemia target cells with a fluorescent dye, rhodamine-123 (Rh-123). The labelling and assay conditions were determined for minimizing spontaneous release (SR). In order to investigate whether NK activity assessed by measuring Rh-123 release agrees with the activity determined by a 51Cr release assay, the NK activity of PBMC was measured simultaneously by both assay methods. Statistical analysis demonstrates that NK activities determined by Rh-123 release correlate well with those measured by 51Cr release. The Rh-123 release assay under the conditions determined was found to be applicable to measurement of the enhanced NK activity resulting from pretreatment of effector leukocytes with interferon-alpha. It is concluded that the Rh-123 release assay with use of K-562 labelled target cells is practical for the assessment of human NK activity in laboratories where use of radioisotopes is not permitted or undesirable.