Effect of antibody to CD134 on perforin-mediated cytolysis in human peripheral blood mononuclear cells.
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.
Li
Ming
M
Department of Rheumatology and Immunology, Shandong Provincial Hospital, Shandong University, Jinan, P.R. China.
Zhang
Yuanchao
Y
D019138:Antibodies, Blocking / Q000502:physiology*; D003602:Cytotoxicity, Immunologic; D015536:Down-Regulation / Q000276:immunology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D053262:Receptors, OX40; D018124:Receptors, Tumor Necrosis Factor / Q000276:immunology*; D016192:Resting Phase, Cell Cycle / Q000276:immunology
D019138:Antibodies, Blocking; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D053262:Receptors, OX40; D018124:Receptors, Tumor Necrosis Factor; C505716:TNFRSF4 protein, human; D054353:Perforin