表达嵌合抗原受体Jurkat细胞株的筛选及功能研究

Objective: It's known that VEGF is over-expressed in almost all kinds of human solid tumors. This research was to design a new gene structure FLT1CAR of chimeric antigen receptor specific to VEGF and to screen the Jurkat cells expressing FLT1CAR which can provide a theoretical basis for future cell therapy.Method:The constructed chimeric antigen receptor gene was inserted into the lentiviral vector named pLV120gn-FLT1CAR .Then Jurkat cells were infected with the virus collected from the 293 T cells which were co-transfected with recombinant vector of plasimd pLV120gn-FLT1CAR, packaging plasimd pCMVdelta8.91 and helper plasimd pMD.G.Finally, we selected the stable transfected cell strains by means of G418 screening and clone selection. To detect the integration and expression of interested gene in the stable transfected cell strains selected,PCR and FCAS was used respectively.The chemotactic effect of the stable transfected cell trend to VEGF was tested by Transwell assay.Results:The agarose gel electrophoresis showed that the constructed lentiviral vector of plasimd pLV120gn-FLT1CAR is correct.PCR and FCAS showed that FLT1CAR has integrated into Jurkat cells and stably expressed. Transwell assay showed that Clone1 and Clone2 from the screened cell strains have obvious chemotactic effect to VEGF. Conclusion: We have Successfully gained the cell clones that express FLT1CAR stably. They have a distinct chemotactic trend to VEGF.