Antigen-binding characteristics of AbCD71 and its inhibitory effect on PHA-induced lymphoproliferation  

作　　者：Ping LEI Yong HE Qing YE Hui-fen ZHU Xiao-mei YUAN Jing LIE Wei XING Sha WO Wei DAI Xin SHEN Guo-bin WANG Guan-xin SHEN 

机构地区：[1]Departments of Immunology and Pharmacology,Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China [2]Departments of Nuclear Medicine , Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China [3]Department of Pathobiology and Physiology, Medicine and Life Science College of Jianghan University, Wuhan 430056, China [4]Departments Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

出　　处：《Acta Pharmacologica Sinica》2007年第10期1659-1664,共6页中国药理学报（英文版）

基　　金：Project supported by grants from the Hi-Tech Research and Development Program of China(№ 2006AA02Z158);Hubei Key Sci &Tech R&D Programme(№ 2006AA301A05)

摘　　要：Aim： To investigate the anti-lymphoproliferative effect of the prepared recombinant chimeric human/murine anti-cluster of differentiation（CD） 71 monoclonal antibody （AbCD71）, which is composed of mouse-derived, antigen-binding variable regions and human-derived constant regions. Methods： After plasmids construction and transfection, the expression of AbCD71 in the transfectoma supernatant was determined by the sandwich ELISA. Indirect immunofluorescence assay was used to measure the antigen-binding characteristic and the percent CD71- expressed peripheral blood mononuclear cells （PBMC）. The antibodies were pufflied from the ascites via diethylaminoethyl（DEAE）-Sephadex A-50 chromatogra- phy and then identified by SDS-PAGE. At last, inhibitory effect of AbCD71 on PHA-induced PBMC proliferation was calculated by methyl thiazolyl tetrazolium （MTT） assay. Results： Constant domain of heavy chain（CH） and light chain（CL）Of AbCD71 were in the human Cy family and human Cκ family, respectively. AbCD71 could compete with its original murine mAb to bind with CD71-positive human leukemia cell line CEM cells. AbCD71 could inhibit the peripheral blood mononuclear cell proliferation induced by phytohemagglutinin（PHA） in vitro in a dosedependent manner, especially at time-points 0 and 12 h after induction. There was no statistical difference when compared with original murine mAb. Conclusion： The AbCD71 is a promising immunosuppressant. Our approach to blocking CD71 with the chimeric human/murine mAb provides a novel strategy for prolonging allograft survival.Aim： To investigate the anti-lymphoproliferative effect of the prepared recombinant chimeric human/murine anti-cluster of differentiation（CD） 71 monoclonal antibody （AbCD71）, which is composed of mouse-derived, antigen-binding variable regions and human-derived constant regions. Methods： After plasmids construction and transfection, the expression of AbCD71 in the transfectoma supernatant was determined by the sandwich ELISA. Indirect immunofluorescence assay was used to measure the antigen-binding characteristic and the percent CD71- expressed peripheral blood mononuclear cells （PBMC）. The antibodies were pufflied from the ascites via diethylaminoethyl（DEAE）-Sephadex A-50 chromatogra- phy and then identified by SDS-PAGE. At last, inhibitory effect of AbCD71 on PHA-induced PBMC proliferation was calculated by methyl thiazolyl tetrazolium （MTT） assay. Results： Constant domain of heavy chain（CH） and light chain（CL）Of AbCD71 were in the human Cy family and human Cκ family, respectively. AbCD71 could compete with its original murine mAb to bind with CD71-positive human leukemia cell line CEM cells. AbCD71 could inhibit the peripheral blood mononuclear cell proliferation induced by phytohemagglutinin（PHA） in vitro in a dosedependent manner, especially at time-points 0 and 12 h after induction. There was no statistical difference when compared with original murine mAb. Conclusion： The AbCD71 is a promising immunosuppressant. Our approach to blocking CD71 with the chimeric human/murine mAb provides a novel strategy for prolonging allograft survival.

关 键 词：AbCD71 chimeric human/murine antibody LYMPHOPROLIFERATION transplantation rejection 

分 类 号：R3[医药卫生—基础医学]