Mesenchymal Stem Cells Overexpressing Interleukin‐35 Propagate Immunosuppressive Effects in Mice

Abstract

Abstract To explore generation of interleukin ( IL )‐35‐expressing mouse adipocyte‐derived mesenchymal stem cells (Ad‐ MSC s) using lentiviral vector and their potential immunosuppressive effects in mice. Ad‐ MSC s were isolated and cultured in vitro and transfected with a lentivirus vector for overexpression of the therapeutic murine IL ‐35 gene. IL ‐35 expression in transfected MSC s ( IL ‐35‐ MSC s) was quantified by enzyme‐linked immunosorbent assay ( ELISA ). The lymphocytes subsets after one‐way mixed lymphocyte culture and in vivo intravenous transplantation were analysed by flow cytometry to evaluate the immunosuppressive effects of IL ‐35‐ MSC s. ELISA was performed to examine IL ‐10, IL ‐17A and IL ‐35 expression in lymphocyte culture. Mouse Ad‐ MSC s were isolated and cultured. IL ‐35 was expressed in the MSC supernatant and serum after IL ‐35 transduction into Ad‐ MSC s by lentiviral vector transfection in vitro and in vivo . The percentage of CD 4 + CD 25 + T regulatory (Treg) cells in mice treated with IL ‐35‐ MSC s significantly increased. IL ‐35‐ MSC s upregulated the CD 4 + CD 25 + Treg cells in the allogeneic mixed lymphocyte reaction system, and lowered the percentage of CD 4 + T cells compared with the other two control groups ( P < 0.01). IL ‐17A expression significantly decreased and IL ‐10 expression significantly increased in IL ‐35‐ MSC s and MSC s when compared by ELISA to the control groups ( P < 0.01). IL ‐35‐transduced Ad‐ MSC s in vivo can enhance proliferation of CD 4 + CD 25 + Treg cells and suppress the function of effector T cells such as T helper (Th) 1, Th2 and Th17 cells and may reduce the development of allograft rejection. Our data suggest that transduced Ad‐ MSC s overexpressing IL ‐35 may provide a useful approach for basic research on cell‐based immunotolerance therapy for inducing transplantation tolerance.