Construction and identification of a drosophila S2 cell line with stoble expression of Mycobacterium tuberculosis-specific Vγ9Vδ2 T cell receptor monomers

Abstract

Objective To construct and identify a drosophila S2 cell line that could stably express Mycobacterium tuberculosis-specific Vγ9Vδ2 T cell receptor (TCR) monomers. Methods Four vectors including the TCR γ9-FB-pMT/V5-His B expression vector, TCR δ2-JB-pMT/V5-His A expression vector, pMT/Bip-BirA expression vector and pCoHygro plasmid were transfected into S2 cells by using calcium phosphate transfection. The S2 cells with stable expression of the Vγ9Vδ2 TCR monomers were screened out with Hygromycin-B. CuSO4 and biotin were used together to induce the expression of biotinylated Vγ9Vδ2 TCR monomers in S2 cells after 4 weeks of culturing. The target proteins in the supernatants of cell culture were identified with dot blot, SDS-PAGE and Western blot assays. Results The results of dot blot, SDS-PAGE and Western blot assays showed that the Vγ9Vδ2 TCR monomers could be stably expressed by the screened S2 cell line. Moreover, the expressed Vγ9Vδ2 TCR monomers were successfully biotinylated. Conclusion The drosophila S2 cell line that could stably express Vγ9Vδ2 TCR monomers was successfully established. This study has paved the way for further investigation on the antigen recognition of γδ T cells and its possible mechanism. Key words: γδT cell receptor; Monomer; S2 cell