Experimental study on the effects of IL-23 in inducing proliferat ion of human peripheral blood mononuclear cell and in killing MUTZ-1 cells in vitro

doi:10.3969/j.issn.1002-7386.2016.20.003

Abstract：Objective To observe the effects of IL -23 in inducing proliferation of human peripheral blood mononuclear cell ( PBMNC) and in killing MUTZ-1 cells—a human myelodysplastic syndrome ( MDS) cell line in vitro. Methods The PBMNCs were separated by ficoll density gradient centrifugation.The experiment was divided into control group (without IL-23), IL-23 high,median,low dose groups (2, 10, 50ngm/l,respectively) in which the PBMNCs were induced by IL-23 for 72 hours in vitro.Then PBMNCs were co-cultured with MUTZ-1 cells.The proliferative ability of PBMNC cells was detected by flow cytometry, and the killing effect of PBMNC on MUTZ-1 cells was measured by LDH releasing method . Moreover the expression levels of IFN-γ, TNF-α, TGF-β, granzyme A, granzyme B and perforin at mRNA and protein levels were detected by Real time-PCR and Western Blot,respectively.Result s After 1, 3, 5,7 -day induction, IL-23 could obviously promote the proliferation of PBMNCs,with a concentration and time-dependent manner, as compared with that in control group ( P <0.05).The positive cell numbers of CD3+, CD4+and CD5+6 were significantly increased, however, CD8+cells were obviously decreased,moreover,the killing effects of PBMNC induced by IL-23 on MUTZ-1 cells were increased. After treated by IL-23, the expression levels of IFN-γ, TNF-α, TGF-β, granzyme A, granzyme B and perforin at mRNA and protein levels were significantly increased , with a concentration and time -dependent manner, as compared with those in control group ( P <0.05).Conclusion IL -23 can obviously promote the proliferation of PBMNCs and can kill MUTZ-1 cells through inducing the expressions of IFN-γ, TNF-α, TGF-β, granzyme A, granzyme B and perforin of PBMNCs.