Abstract
CD134 (OX40) is a new member of the tumor necrosis factor receptor (TNFR) superfamily. The influence of CD134 on perforin-mediated cytolysis remains obscure. In this study, human peripheral blood mononuclear cells (PBMC) were induced with phytohemagglutinin (50 microg/mL), by using anti-CD134 blocking monoclonal antibody (MAb; 1, 5, and 10 microg/mL) for 6, 12, 24, and 48 h. Perforin-mediated cytolysis against human erythrocytes of PBMC was detected. The expression of perforin mRNA and protein was also measured by reverse transcription-polymerase chain reaction and flow cytometry. Our data showed that the perforin-mediated cytolysis of PBMC was downregulated by various concentrations of anti-CD134 blocking MAb for different times and reached a minimum at 24 h at any concentration. Anti-CD134 blocking MAb induced downregulation of perforin-mediated cytolysis of PBMC in a dose-dependent manner in the range of 1-5 microg/mL. Perforin- mediated cytolysis of PBMC reached a plateau when the concentration of anti-CD134 blocking MAb exceeded 5 microg/mL. Anti-CD134 blocking MAb also induced the inhibition of the expression of perforin protein and mRNA in PBMC. In conclusion, anti-CD134 blocking MAb inhibits perforin-mediated cytolysis of PBMC by, at least in some part, decreasing the production of preforin, which may be useful in treating several autoimmune diseases.
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