Preparation and biological identification of a functional monoclonal antibody,mouse anti-human B and T lymphocyte attenuator

To prepare a functional monoclonal antibody of mouse anti-human B and T lymphocyte attenuator(BTLA),highly expressed human BTLA transfectant L929/BTLA was used as immunogen to immunize 6~8 weeks BALB/c mice and then the mouse splenic cells were fused with mouse plasmocytoma cells SP2/0.The hybridoma cells were screened with another human BTLA transfectant 293T/BTLA.First strip analysis was performed to identify immunoglobulin subclass of the produced monoclonal antibodies,and the inhibitory effect on T cell proliferation induced by anti-CD3 mAb of such monoclonal antibodies was studied by MTT assay.The results showed that this newly generated mAb 8H9 could specifically recognize the BTLA protein expressed on these transfected cell lines,and could bind with not only resting but also activated T lymphocytes.Moreover,this monoclonal antibody could mediate obviously an inhibitory effect on the proliferation of T cells.