Establishment of cell line transfected with human BTLA gene and preliminary study on its biological function

    WANG Xueª²feng, CHEN Yongª²jing, WANG Qin, YANG Keª²fang, DAI Qun, WANG Fengª²ming, ZHANG Xueª²guang*

    Institute of Medical Biotechnology, Soochow University, Key Laboratory of Stem Cell of Jiangsu Province, Suzhou 215007, China

    [Abstract] AIM: To establish 293T cells expressing human BTLA gene, a novel attenuator for B and T lymphocyte. METHODS: The gene encoding human BTLA was amplified by RTª²PCR from PHA activated T cells, and then the target gene fragment was inserted into retrovirus vector pEGZª²Term after being digested with EcoR I and BamH I. The recombinant vector was transfected into 293T cells with LipfectAMINETM 2000, and the cells was further selected with Zeocin. Effect of 293T/BTLA cells on T cells proliferation and activation in vitro was been studied by methods of MTT and cytometry. RESULTS: The stable expression of human BTLA on the transfected cell line was identified by flow cytometry analysis. In vitro, 293T/BTLA cells had an inhibitory effect on the proliferation of T cells stimulated by antiª²CD3 mAb and downª²regulated the expression of activation marker CD25 on the surface of T cells and decreased the secretion of IFNª²¦Ã and ILª²10. CONCLUSION: A cell line 293T/BTLA stably expressing the human BTLA protein has been obtained and it can partially inhibit the proliferation and activation of T cells in vitro. ......