Quantitative analysis of the 51Cr release cytotoxicity assay for cytotoxic lymphocytes

Abstract

Using 51Cr-labelled P-815 mastocytoma cells as target cells and CS7BL/6 spleen cells sensitized against DBA/2 antigens as effector cells, it is shown that the variation in the observed specific 51Cr release over a broad range of experimental conditions can be explained on the basis of a simple physical model of the interaction process. The model assumes that a target cell can be destroyed only after contact with an effector cell, contact takes place on a random basis, one contact is sufficient, and that one effector cell can kill several targets with unchanged efficiency. The fraction of target cells destroyed ( f) depends only on the incubation time ( t), the number of effector cells ( n) and a constant interaction probability (δ). Thus f = 1 − e − nδt . However, the experimental measurement, the fraction of 51Cr specifically released into the supernatant during the assay, may not be the same as the fraction of target cells destroyed because it takes considerable time for the releasable 51Cr to be released from a damaged target cell. This can be overcome experimentally by following the standard 37 °C incubation with a further incubation at 45 °C during which there are no new lytic events but all previously damaged target cells release the remainder of their releasable 51Cr. The model enables one to obtain accurate measurements of relative effector cell frequency over a broad range of experimental conditions.