Regulation by glutathione of the effect of lymphokines on differentiation of primary activated lymphocytes. Influence of glutathione on cytotoxic activity of CD3-AK-.
By activating murine lymphocytes with anti-CD3 antibodies for 1 to 2 days, we generated a subset of activated killer cells, namely CD3-AK-. CD3-AK- mediated the slow lysis (20-h 125I-UdR release assay) of allogeneic P815 but had little effect on syngeneic HFL/b cells. Addition of IL-2 (murine or human) or an IL-2 inducer such as PMA in the assay medium induced the cytolytic activity of CD3-AK- on HFL/b. The activating effect of murine IL-2 and PMA on CD3-AK- was decreased by anti-murine IL-2 mAb. Although anti-murine IL-4 mAb alone did not show any effect, it enhanced the inhibitory effect of anti-IL-2 mAb, suggesting that IL-2 and IL-4 may have a synergistic effect on the cytolytic activity of CD3-AK-. Incubation of CD3-AK- with L-buthionine-(SR)-sulfoximine (BSO), an inhibitor of de novo glutathione (GSH) synthesis, decreased cellular GSH levels and inhibited the cytolytic activity of CD3-AK-, in a concentration-dependent manner. This inhibitory effect of BSO was not primarily due to a general cytotoxic effect and was positively correlated with the requirement for IL-2 for the CD3-AK(-)-mediated killing of the target cells. Incubation of CD3-AK- with GSH or 2-ME, which increased the level of cellular GSH, reversed the inhibitory effect of BSO. These results suggest that cellular GSH may regulate the effect of lymphokine(s) such as IL-2 and thus affect the differentiation of activated primary cytotoxic lymphocytes.
Liang
S M
SM
Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892.
Liang
C M
CM
Hargrove
M E
ME
Ting
C C
CC
D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D002454:Cell Differentiation; D003602:Cytotoxicity, Immunologic / Q000502:physiology; D005260:Female; D005978:Glutathione / Q000502:physiology*; D007376:Interleukin-2 / Q000502:physiology*; D015847:Interleukin-4 / Q000502:physiology*; D007694:Killer Cells, Natural / Q000166:cytology*; D008213:Lymphocyte Activation; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D011948:Receptors, Antigen, T-Cell
D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D007376:Interleukin-2; D011948:Receptors, Antigen, T-Cell; D015847:Interleukin-4; D005978:Glutathione