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《Current Immunology》 2014-01
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Construction and functional analysis of a mouse secretory TIGIT lentiviral vector

CHEN Xi;GAO Yi;HUANG Min;AIHAITI Aziguli;CHEN Zhong-hua;YV Xue-feng;HE Wen-tao;Department of Endocrinology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology;Institute of Organ Transplantation,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology;Department of Laboratory Medicine,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology;  
Our aims are to construct a lentiviral vector Lenti-TIG-Fc which encodes the extracellular domain of mouse TIGIT and the human IgG-Fc fusion gene,and to verify the functions of the fusion protein TIG-Fc and lay a foundation for the further study in immune regulation.The extracellular domain of mouse TIGIT and human IgG-Fc fusion gene was cloned into plasmid pLOX.Correctly connected cloning TIG-Fc-pLOX was identified and then co-transfected into HEK 293T cells together with pCMVAR8.2 and pMD.G,The supernatant containing lentivirus Lenti-TIG-Fc was harvested.293T cells infected hy the Lenti-TIG-Fc were co-cultured with LPS-induced mature dendritic cells(DCs) from BALB/c mice.IL-10 level secreted by DCs was detected by ELISA.The results showed that TIG—Fc fusion gene was successfully cloned and inserted to lentivirus plasmids.293T cells infected by the Lenti-TIG-Fc consistently secret TIG-Fc fusion protein and could significantly promote secretion of IL-10 by mature DCs.
【Fund】: 国家自然科学基金(81102260);; 中央高校基本科研业务费资助(HUST2012QN190)
【CateGory Index】: R392
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