The Study About ScTCR/multimer Identifing Xuanwei Lung Adenocarcinoma XWLC-05 Tansfected With HLA-A*0201

[Objective]Among the HLA-A, B, DRB1, DQB1 alleles of Yunnan human lung cancer, the frequency of HLA-A*02 is significantly higher. The data manifest that HLA-A2 can present wild P53_264-272 peptide, and there is a soluble single-chain T Cell Receptor(scTCR) that can identify this P53_264-272 /HLA-A*0201 complex specificially. The multimer of this TCR(264scTCR/multimer) especially its tetramer maintain the highest specificity. In this study, we observeed specific identification of the 264scTCR/multimer with xuanwei lung adenocarcinoma which transfected with HLA-A*0201 and provided dependable theories for specific killing of the CD8⁺CTLs which presented this kind of TCR.[Methods]We selected xunwei lung adenocarcinoma XWLC-05 expressing HLA-A*0201 negatively by Polymerase Chain Reaction-Sequence Specific Primers(PCR-SSP) for this experiment and then transfected XWLC-05 with pcDNA-3．1-HLA-A*0201 using liposomes. After XWLC-05 expressed HLA-A*0201 positively, we measured weather it can be identified by the 264scTCR/multimer.[Results]Transfected XWLC-05 with pcDNA-3．1-HLA-A*0201 and aquire stable positive transfected cell clone by G418．After pulsed with wild P53_264-272 peptide andβ2-microglobulin at 37℃water bath. Cells were incubated with TCR P53-264-PE for FCM.The result was positive.[Conclusion]There was correlation between HLA-A*0201 gene and high frequency of Yunnan lung cancer. XWLC-05 which was transfected with pcDNA-3．1-HLA-A*0201 could espress efficiently and presented wild P53_264-272 , and this P53_264-272 /HLA-A*0201 complex could be identified by 264scTCR/multimer. This provided the possibility that T cells which precented 264scTCR could kill the HLA-A*0201 positive expressing lung cancer specificially and established foundation of the P53 therapy.