Construction and biological identification of the transfectant cells expressing ??TCR with specific CDR3 sequence
- VernacularTitle:特异性CDR3区取代型??TCR转染细胞的构建及生物学功能鉴定
- Author:
Xueyan XI
;
Wei CAO
;
Lianxian CUI
;
Wei HE
- Publication Type:Journal Article
- Keywords:
??T cell;
??TCR with specific CDR3 sequence;
Antigen recognition
- From:
Chinese Journal of Immunology
2001;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct of transfectant cells expressing ??TCR with specific CDR3 sequence.Methods:Specific CDR3 region sequence of DBS4.3,a known ??T cell clone,was inserted into ?9 and ?2 chain to substitute original CDR3 sequence using overlapping PCR method.After the full-length ?9 and ?2 chains were ligated with expression vector pREP7and pREP9 respectively,they were co-trasfected into a cell line of human Jurkat cells with TCR ? chain gene-defective mutant J.RT3-T3.5.When the transfectant cells expressing specific ??TCR were stimulated by antigen,production of IL-2 was detected by ELISA and Realtime PCR.Results:By ELISA and Realtime PCR,it was exhibited that the transfectant cells expressing DBS4.3 specific ??TCR secreted IL-2 under stimulation of iso-butylamine and anti ??TCR antibody.Conclusion:The transfectant Jurkat cells expressing ??TCR with specific CDR3 sequence are successfully constructed.It provides a platform for the research of recognition mechanism of ??TCR.
