Allogeneic lymphocyte cytotoxicity (ALC) in rats: establishment of an in vitro assay, and direct evidence that cells with natural killer (NK) activity are involved in ALC.

The evidence that NK cells can recognize and kill allogeneic lymphocytes has hitherto been based mainly on experiments in intact animals. Here we report results from an in vitro assay, showing allogeneic lymphocyte cytotoxicity in cell suspensions enriched for NK activity against tumour cells by Percoll gradient centrifugation of nylon-wool non-adherent cells. The addition of phytohaemagglutinin (PHA) to the NK-target cell cultures greatly enhanced the cytotoxic response against K562 and allogeneic, but not syngeneic, lymphocytes. The effector cells of ALC are present in the spleen of both euthymic and athymic nude rats, and to a lesser extent in the blood. ALC is augmented by interferon pretreatment of the effector cells, and by depleting the effector cell suspensions of all T cells and helper T cells with the monoclonal antibody MRC Ox19 and W3/25, respectively. Conversely, the activity was nearly abolished by depleting the cell suspensions of MRC Ox8+ cells reacting with rat cytotoxic T cells and NK cells. Furthermore, removal of residual B cells (Ox12+ cells) from the effector cells or attempts to block any putative antibody-dependent cellular cytotoxic mechanism in vitro with the monoclonal antibody Ox12 did not inhibit the NK activity against allogeneic lymphocytes nor against tumour cells. ALC in vitro did not discriminate between T and B or large and small lymphocyte targets. These characteristics of the ALC effector cells substantiate that they are present within the thymus-independent population of cells with NK activity, and are dependent on neither B cells nor immunoglobulin for their recognition and destruction of the target.