TY - JOUR
T1 - Inhibition of cell-mediated cytotoxicity by 2-cyclohexene-1-one
T2 - Evidence for a role for glutathione and/or glutathione-protein interactions in cytolysis
AU - MacDermott, R. P.
AU - Bertovich, M. J.
AU - Bragdon, M. J.
AU - Nash, G. S.
AU - Leusch, M. S.
AU - Wedner, H. J.
PY - 1986
Y1 - 1986
N2 - In order to explore the role of glutathione in cell-mediated cytotoxicity, we have examined the effect of the sulphydryl-reactive and glutathione-depleting agent 2-cyclohexene-1-one on antibody-dependent cellular cytotoxicity, spontaneous cell-mediated cytotoxicity, and cell-mediated lympholysis by human peripheral blood mononuclear cells. 2-Cyclohexene-1-one significantly inhibited (P < 0.001) both antibody-dependent and spontaneous cell-mediated cytotoxicity using three different cell-line targets, at three different killer:target cell ratios (10:1, 25:1 and 50:1). Using K-562 cell-line targets, spontaneous cell-mediated cytotoxicity was inhibited by 2-cyclohexene-1-one with an ID50 of 0.71 x 10-4 M - 1.48 x 10-4 M, while antibody-dependent cellular cytotoxicity was less sensitive to inhibition, and required slightly higher concentrations of 1.48 x 10-4 M - 3.98 x 10-4 M to achieve 50% inhibition. Similar results were seen with human colon tumor cell-line and Chang liver cell-line cells as targets. Maximal inhibition occurred when 2-cyclohexene-1-one was added to the cytotoxicity assay 60 min prior to, at the start of, or within the first 60 min of a 4-hr assay; inhibition of cytotoxicity occurred with pretreatment of effector cells; and no inhibition of cytotoxicity was observed with pretreatment of target cells. Both the allogeneic mixed leucocyte reaction and cell-mediated lympholysis were also significantly inhibited (P < 0.001) by 2-cyclohexene-1-one. These studies demonstrate that 2-cyclohexene-1-one is an effective inhibitor of cell-mediated cytotoxicity and suggest that glutathione, specific glutathione-protein interactions, or protein-bound sulphydryl groups are involved in allowing cells to carry out cytolysis.
AB - In order to explore the role of glutathione in cell-mediated cytotoxicity, we have examined the effect of the sulphydryl-reactive and glutathione-depleting agent 2-cyclohexene-1-one on antibody-dependent cellular cytotoxicity, spontaneous cell-mediated cytotoxicity, and cell-mediated lympholysis by human peripheral blood mononuclear cells. 2-Cyclohexene-1-one significantly inhibited (P < 0.001) both antibody-dependent and spontaneous cell-mediated cytotoxicity using three different cell-line targets, at three different killer:target cell ratios (10:1, 25:1 and 50:1). Using K-562 cell-line targets, spontaneous cell-mediated cytotoxicity was inhibited by 2-cyclohexene-1-one with an ID50 of 0.71 x 10-4 M - 1.48 x 10-4 M, while antibody-dependent cellular cytotoxicity was less sensitive to inhibition, and required slightly higher concentrations of 1.48 x 10-4 M - 3.98 x 10-4 M to achieve 50% inhibition. Similar results were seen with human colon tumor cell-line and Chang liver cell-line cells as targets. Maximal inhibition occurred when 2-cyclohexene-1-one was added to the cytotoxicity assay 60 min prior to, at the start of, or within the first 60 min of a 4-hr assay; inhibition of cytotoxicity occurred with pretreatment of effector cells; and no inhibition of cytotoxicity was observed with pretreatment of target cells. Both the allogeneic mixed leucocyte reaction and cell-mediated lympholysis were also significantly inhibited (P < 0.001) by 2-cyclohexene-1-one. These studies demonstrate that 2-cyclohexene-1-one is an effective inhibitor of cell-mediated cytotoxicity and suggest that glutathione, specific glutathione-protein interactions, or protein-bound sulphydryl groups are involved in allowing cells to carry out cytolysis.
UR - http://www.scopus.com/inward/record.url?scp=0022588931&partnerID=8YFLogxK
M3 - Article
C2 - 2937720
AN - SCOPUS:0022588931
SN - 0019-2805
VL - 57
SP - 521
EP - 526
JO - Immunology
JF - Immunology
IS - 4
ER -