IN VITRO CO-STIMULATORY ACTIVITY OF HUMAN B7.2(IgV+C) PROTEIN PRODUCED BY ENGINEERED BACTERIA

作     者：闫晓彩 司履生 王一理 刘培军 来宝长 耿宜萍 

作者机构：Institute of Immunopathology Med. Sch. of Xi'an Jiaotong Univ. Xi'an 710061 China 

出 版 物：《Academic Journal of Xi'an Jiaotong University》 (西安交通大学学报（英文版）)

年 卷 期：2001年第13卷第1期

页      面：16-19页

学科分类：1001[医学-基础医学(可授医学、理学学位)] 100102[医学-免疫学] 10[医学] 

基　　金：This work was supported by the National Natural Science Foundation of China(No. 39470293) 

主　　题：human B7. 2/CD86 co-stimulation fusion protein gene expression 

摘      要：Objective To express human B7. 2 extracellular domain with prokaryote expression system and to evaluate its biological activity in vitro. Methods PCR was used to amplify the extracellular region of human B7. 2 which contained both the IgV and IgC domains. The recombinant PGEX-4T-3/hB7. 2 (IgV+C) was obtained by cloning the PCR product into a prokaryote expression plasmid PGEX-4T-3 and was transformed into the host strain of DH5-a. Tke fusion protein consisted of GST and hB7. 2(IgV+C) was identified by SDS-PAGE and Western blotting. T cell activation was observed by exposing purified T lymphocytes to the fusion protein and [3H]-TdR incorporation with the presence of the first signal imitated hy anti-CD3 antibody. Results The fusion protein GST-hB7. 2 (IgV+ C) was produced and detected in inclusive body form from engineered bacterial cells. With the first signal existed,T lymphocytes proliferated when it was co-stimulated by the fusion protein. Conclusion These results indicated that the functional human B7. 2(IgV+C) fusion protein can be produced in bacterial cells and the fusion protein displays the co-stimulatory activity in T lymphocytes activation.