Title: Effect of CD86 on the differentiation of CD8~+T cell
Abstract: Objective To investigate the effect of CD86 (B7-2) on the differentiation of CD8 +T cell in vitro. Methods CD86 gene was cut out from plasmid pCDM8 by restriction endonuclease Xho Ⅰ. It was inserted into eukaryotic expression vector pCDNA3 and identified by restriction enzyme BamH Ⅰ. Further, the recombinant plasmid vector pCDNA3-CD86 was transferred into human hepatocellular carcinoma cell line HMC7721 with cationic liposome. After selected by 600 μg/mL G418, the G418-resistant clone expressing CD86 stably and highly was isolated by flow cytometry (FCM). Finally, human peripheral blood mononuclear cells (PBMCs) were separated from peripheral blood of healthy volunteer, and co-cultured with target cells at E/T ratio of 20. 48 h later, CD3 +T cells were assayed for the secretion of the intracellular cytokines IL-4 and IFN-γ using FCM. Results The recombinant plasmid vector pCDNA3-CD86 was constructed successfully. The CD86 expression rate in HMC7731-CD86 cell was 30.8%, but the rate in HMC7721 cell was 0.98%. The expression rate of IL-4 and IFN-γ were 1.92% and 24.4% in the CD3 +T cells of healthy volunteer. After PBMCs were co-cultured with target cells for 48 h, whether target cells were stimulated by IFN-α or not, the positive rate of IL-4/IFN-γ on CD3 +T cells was more than 1 in the cell line HMC7721-CD86 groups. In contrast, the positive rate of IL-4/IFN-γ was less than 1 in the cell line HMC7721 groups. Conclusions In cell culture, CD86 can induce CD8 +T cell activation and regulates Tc2 phenotype differentiation.
Publication Year: 2003
Publication Date: 2003-01-01
Language: en
Type: article
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