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《Current Immunology》 2011-01
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The costimulatory effect of mouse B7-H3 gene in the activation of T cells

SUN Jing1,2,YAN Ru-hong1,3,GU Wen-chao1,FU Feng-qing1,ZHANG Guang-bo1,CHEN Xu-qin1,ZHOU Ying-hui1,ZHANG Xue-guang1(1.Institute of Biotechnology,Soochow University,2.Suzhou Health Technology College,3.Clinical Diagnosis Laboratory of the Second Affiliated Hospital,Soochow University,Suzhou 215007,China)  
The biological function of mouse B7-H3 was still not identified yet.This paper was aimed to generate an engineered CHO cell line harboring mouse B7-H3 and to investigate its biological function and effects on T lymphocyte activation.The genes coding mouse B7-H3 were obtained from mouse liver cDNA by RT-PCR and then the target gene fragment was inserted into eukaryotic vector pIRES2-EGFP after being digested with EcoR Ⅰ and Bgl Ⅱ to construct the recombinant vector pIRES2-EGFP/B7-H3.This recombinant vector was transfected into CHO cells with LipfectAMINETM 2000 and followed by G418 selection.RT-PCR,Western blotting and flow cytometry(FCM) were used to detect the expression of B7-H3.Effects of the transfected cells on T cells proliferation and cytokine production in vitro was studied by methods of CCK8 and ELISA.In these ways,a stable cell line constitutively expressing the mouse B7-H3 was established successfully,and the mouse B7-H3 could obviously promote the proliferation of T cells in and secretion of IL-2 and IFN-γ in vitro.This effect could be partly blocked by mAb.It is evident that the engineered mouse B7-H3/CHO cell line provides a gain of function cell model for the further understanding of the biological role of B7-H3.
【Fund】: 国家自然科学基金资助项目(31040022、30771958);; 苏州市社会发展基金资助项目(ss08016)
【CateGory Index】: R392.12
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