Induction of lymphokine synthesis in peripheral blood mononuclear cells with phorbol ester and calcium ionophore allows precise measurement of individual variations in capacity to produce IL 2.
Absolute capacity for IL2 production by human peripheral blood mononuclear cells (PBMC) was studied using 12-O-tetradecanoylphorbol 13-acetate (TPA) and calcium ionophore A23187, which act synergistically, to induce lymphokine synthesis. Culture parameters were optimized for the TPA/A23187 stimulation such that maximal IL2 titers were produced with a high degree of reproducibility. Thus, using a synthetic medium, TPA/A23187 at 20/50ng/ml respectively, and a cell concentration of 2.5 X 10(6)/ml, IL2 titers in the cultures increased linearly over a period of 96h, reaching values at least 15-fold higher than with lectin stimulation. This allowed for determinations of IL2 synthetic capacity in individual blood samples. Large fluctuations in normal IL2 production (range 1775-10654 BRMP U/ml at 48h) were observed among 23 normal persons. A statistically significant lower IL2 productive capacity was observed in the age group above 40 as compared to those under 40. The lower rates of IL2 synthesis in a group of patients with Hodgkin's disease was seen only among those who had undergone immunosuppressive therapy; newly diagnosed cases fell within the normal range.
D000001:Calcimycin / Q000494:pharmacology; D006689:Hodgkin Disease / Q000276:immunology; D006801:Humans; D066298:In Vitro Techniques; D007376:Interleukin-2 / Q000096:biosynthesis*; D007962:Leukocytes / Q000276:immunology*; D008222:Lymphokines / Q000096:biosynthesis*; D010835:Phytohemagglutinins / Q000494:pharmacology; D012016:Reference Values; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D007376:Interleukin-2; D008222:Lymphokines; D010835:Phytohemagglutinins; D000001:Calcimycin; D013755:Tetradecanoylphorbol Acetate