Estimation by limiting dilution analysis of human IL 2-secreting T cells: detection of IL 2 produced by single lymphokine-secreting T cells.

摘要: We present here a culture method for the estimation, in human blood, of number lymphocytes that can respond to mitogen by producing interleukin 2 (IL 2). T cells are cultured at limiting dilutions with PHA or Con A presence Epstein Barr virus-transformed lymphoblastoid (EB-LCL), and supernatants tested 3 days later IL content cell proliferation assay. The distribution negative wells follows expected Poisson "single-hit" relationship, suggesting assay is sensitive single type. On average, 16.3% peripheral blood mononuclear produce such clonal cultures (mean 12 determinations; SD = 5.6%). Surprisingly, irradiation (up 2000 rad) titrated responder population diminishes estimated frequencies less than 50%. ability detect levels containing only single, nonproliferating lymphocyte allows us estimate amount generated an individual effector during 3-day interval after stimulation. average responding, irradiated generates 0.92 pg (median) within days. presented provides straightforward way provide independent estimates responding lymphokine production per variety clinical situations.