Establishment of cell line transfected with human GL50 gene and preliminary study on its co-stimulatory effect on T cell activation and proliferation

Objective: To establish L929 cell expressing human GL50 gene and analyze its co-stimulatory effect on T cell activation and proliferation in vitro.Methods: The gene encoding human GL50 was amplified by RT-PCR from human B lymphoma Daudi,and then the target gene fragment was inserted into eukaryotic vector pIRES2-EGFP after being digested with EcoR I and BamH I.The recombinant vector pIRES2-EGFP-GL50 was transfected into L929 cells with Lipfectamine 2000,and the cells were further selected with G418 for a long time.GL50 expressed on L929/GL50 cells was analyzed by FCM.Effect of L929/GL50 cells on T cells proliferation and cytokine production in vitro was studied by methods of CCK-8 and cytometry.Results: L929/GL50 stably expressing human GL50 was successfully established.In vitro,L929/GL50 cells could obviously promote the proliferation of T cells,while up-regulating the production of IL-4,IL-10 and IL-17 measured by ELISA.Conclusion: The signal of ICOS/GL50 can play an important role in immune response.A cell line L929/GL50 has been obtained and it can provide the material basis for further research of the function of ICOS/GL50.