Experimental study of inhibition of ZGDHu-1 on T lymphocytes activation
Lian-Nü Qiu, Yonglie Zhou
Abstract
Lian-Nü Qiu, Yonglie Zhou
Abstract
Objective To investigate the effects of ZGDHu-1 on T lymphocytes activation in vitro to elucidate its immunosuppressive effects. Methods Lymphocytes isolated from healthy persons were stim-ulated with phytohanmagglutinin(PHA) and different experimental groups were set by cocultured for 24 h, 48 h with ZGDHu-1 or with ZGDHu-1 and Cyclosporin A(CSA). To assess the proliferation and apoptosis of T lymphocytes, we detected CD3~+ CD69~+ , CD3~+ CD25~+ , CD4~+ CD25~+ , CD8~+ CD25~+ and CD3~+ Fas~+, CD4~+ Fas~+ , CD8~+ Fas~+ with flow cytometry. The early apoptosis rate of lymphocytes was analyzed with flow cytometry. Culture supernatant IL-2 and TGF-β1 were detected with ELISA. Results ZGDHu-1 decreased PHA activative CD3~+ CD69~+, CD3~+ CD25~+, CD4~+ CD25~+ and CD3~+ Fas~+, CD4~+ Fas~+, Annexin V~+/ PI~- and inhibited IL-2 secretion and promoted TGF-β1 secretion respectively. ZGDHu-1 has synergistic effect with CSA to be more obvious. Conclusion ZGDHu-1 can inhibit T lymphocytes activation and de-creased apeptosis of T lymphocytes. ZGDHu-1 has synergistic effect with CSA to be obvious. Key words: ZGDHu-1; Cyclosporin A; T cells activation; Apoptosis; Flow cytometry
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Objective To investigate the effects of ZGDHu-1 on T lymphocytes activation in vitro to elucidate its immunosuppressive effects. Methods Lymphocytes isolated from healthy persons were stim-ulated with phytohanmagglutinin(PHA) and different experimental groups were set by cocultured for 24 h, 48 h with ZGDHu-1 or with ZGDHu-1 and Cyclosporin A(CSA). To assess the proliferation and apoptosis of T lymphocytes, we detected CD3~+ CD69~+ , CD3~+ CD25~+ , CD4~+ CD25~+ , CD8~+ CD25~+ and CD3~+ Fas~+, CD4~+ Fas~+ , CD8~+ Fas~+ with flow cytometry. The early apoptosis rate of lymphocytes was analyzed with flow cytometry. Culture supernatant IL-2 and TGF-β1 were detected with ELISA. Results ZGDHu-1 decreased PHA activative CD3~+ CD69~+, CD3~+ CD25~+, CD4~+ CD25~+ and CD3~+ Fas~+, CD4~+ Fas~+, Annexin V~+/ PI~- and inhibited IL-2 secretion and promoted TGF-β1 secretion respectively. ZGDHu-1 has synergistic effect with CSA to be more obvious. Conclusion ZGDHu-1 can inhibit T lymphocytes activation and de-creased apeptosis of T lymphocytes. ZGDHu-1 has synergistic effect with CSA to be obvious. Key words: ZGDHu-1; Cyclosporin A; T cells activation; Apoptosis; Flow cytometry
Key concepts: Flow cytometry, IL-2 receptor, CD3, Annexin, CD8, Apoptosis, Molecular biology, CD69