Aim: To establish 293T cells expressing human BTLA gene, a novel attenuator for B and T lymphocyte.
Methods: The gene encoding human BTLA was amplified by RT-PCR from PHA activated T cells, and then the target gene fragment was inserted into retrovirus vector pEGZ-Term after being digested with EcoR I and BamH I. The recombinant vector was transfected into 293T cells with LipfectAMINE 2000, and the cells was further selected with Zeocin. Effect of 293T/BTLA cells on T cells proliferation and activation in vitro was been studied by methods of MTT and cytometry.
Results: The stable expression of human BTLA on the transfected cell line was identified by flow cytometry analysis. In vitro, 293T/BTLA cells had an inhibitory effect on the proliferation of T cells stimulated by anti-CD3 mAb and down-regulated the expression of activation marker CD25 on the surface of T cells and decreased the secretion of IFN-gamma and IL-10.
Conclusion: A cell line 293T/BTLA stably expressing the human BTLA protein has been obtained and it can partially inhibit the proliferation and activation of T cells in vitro.