Construction and expression analysis of idiotype TCR Vβ2 DNA vaccine in vitro

AIM:To develop an anti-lymphoblastic leukemia TCR idiotypic DNA vaccine, analyze its transfer activity into K562 cells and to detect its expression in vitro. METHODS:The TCR Vβ2 gene segment, which was identified from an idiotypic TCR Vβ2 clone-Molt4 cell line, was amplified using RT-PCR, and the PCR products were then cloned into pIRES vector. The recombinant plasmids were transferred into K562 cells. The condition of idiotypic protein expression was tested by indirect immunophenotyping fluorescein dyeing, SDS-PAGE and Western blotting. RESULTS:The recombinant DNA plasmid, pIRES-TCR Vβ2, was developed successfully. The expression of TCR Vβ2 was identified on the surface of K562 cells. A 15 kD protein, which bound to TCR Vβ2 antibody specifically, were identified from pIRES-TCR Vβ2 transfected K562 cells by Western blotting, indicating that TCR Vβ2 protein was expressed in vitro. CONCLUSION:The recombinant plasmid pIRES-TCR Vβ2 DNA vaccine was developed successfully, which was expressed TCR Vβ2 protein specifically in transfected K562 cells.