Study on polarized Th1/Th2 cell in vitro from adult human peripheral blood |
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Authors: | LI Zhen ZENG Yao-ying HE Xian-hui CAI Xiao-chang ZENG Shan DI Jing-fang |
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Institution: | 1. Department of Dermatology, the First Affiliated Hospital of Medical College, Jinan University, Guangzhou 510632. China;
2. Institute of Tissue Transplantation and Immunology, Jinan University, Guangzhou 510632. China |
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Abstract: | AIM: To investigate the details of CD4+ T cell polarized to Th1/Th2 in vitro. METHODS: After isolated the PBMCs and blood-plasma from adult human peripheral blood by Ficoll-Hypaque centrifugation, the PBMC culture procedure with or without the self-blood -plasma was applied to polarize T cells in vitro, these cells were polarized by PHA(20 mg/L),non-PHA respectively. The polarized rates of Th cell after 24 h,48 h,72 h were estimated respectively by flow cytometry following two-color immunofluorescent staining. RESULTS: CD4+T cell would polarize to Th1/Th2 two subsets after self-cytokines and PHA activation in vitro. The polarized rates of T cell after cultured for 24 h,48 h and 72 h were (13.28%±1.59%)/(12.70%±1.65%),(17.19%±1.03%)/(17.50%±1.30%),(19.49%±2.87%)/(18.58%±1.49%) respectively, but the polarized rates of T cell were very low if without self-blood-plasma. The difference between them was significant. The ratios of Th1/Th2 cells were about 1. CONCLUSION: CD4+T cell from adult human peripheral blood would polarize to Th1/Th2 two subsets in the presence of self-blood-plasma and PHA(20 mg/L) in vitro, and the cell number of Th1 and Th2 would be in balance. |
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Keywords: | T-lymphocytes Chemotactic factors Flow cytometry |
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