Comparison of the activated lymphocyte into which shPTPN3 was introduced and the activated lymphocyte into which a control vector was introduced

Immunotherapy using activated lymphocytes has recently been developed, but there is not yet the biomarker which can predict curative effect. In our recent study, we found that random migration is significantly increased in activated lymphocyte. Furthermore, it is suggested that random migration may affect cytotoxicity. Therefore we considered that migration of lymphocyte may become the effect prediction biomarker of Immunotherapy. And then we analyzed the gene which fluctuated by lymphocyte activation to search the molecules which regulate migration of lymphocyte.As the result, we found out that expression of PTPN3 fluctuates before and after activation of the lymphocyte.Next, we compared the gene expression of the activated lymphocyte into which shPTPN3 was introduced and of the activated lymphocyte into which a control vector was introduced to search the action mechanism of PTPN3. We prepared human peripheral blood mononuclear cells (PBMCs) and cultured PBMCs in RPMI medium supplemented with 200 U/ml interleukin(IL)-2 in culture plates coated with 2.5 ug/ml monoclonal antibody to CD3 for five days to activate lymphocytes. The non-attached fraction was collected and divided 2 groups.And then shPTPN3 was transfected into 1 group by using lentivirus and control vector was transfected into the other equally. We compared the gene expression of two groups to search the action mechanism of PTPN3.