Effect of interleukin 12A on proliferation and differentiation of CD4+CD25+Foxp3+Treg

doi:10.12092/j.issn.1009-2501.2019.05.003

Abstract

Abstract:

AIM: To study the role of interleukin 12A (IL-12A) in promoting proliferation and differentiation of CD4+CD25-T cells into CD4+CD25+Foxp3+Treg（iTreg）. METHODS: CD4+CD25-T cells in umbilical cord blood were selected by immunomagnetic beads. Both the experimental group and the control group were activated by anti-CD3 and anti-CD28 monoclonal antibodies, and induced by adding 10 ng/mL of TGF-1, while the experimental group was simultaneously induced by adding 5 ng/mL of IL-12A.After the cells were cultured for 72 h, the ratio of CD4+CD25+Foxp3+Treg (iTreg) cells was detected by flow cytometry (FACS). The expression of Foxp3, Smad2, Smad3 and Smad6 in the cells was detected by protein immunoblotting (Western-Blot), and the expression of TGF-β1 was detected by real time fluorescence quantitative PCR.iTreg and CD4+CD25-T cells were induced by co-culture in vitro, and lymphocyte activity was assessed by CCK-8.The co-culture of 5 ng/mL IL-12A with iTreg and CD4+CD25-T cells in vitro was used to detect the effect of IL-12A and iTreg on the activity of CD4+CD25-T cells.RESULTS:IL-12A could promote the differentiation of CD4+CD25-T into iTreg by FACS detection. CCK-8 showed that iTreg could inhibit the activity of CD4+CD25-T cells in a concentration-dependent manner. IL-12A can synergize the inhibitory effect of iTreg on CD4+CD25-T cell activity.Western-Blot and RT-QPCR results showed that Foxp3, Smad2, Smad3 and Smad6 expression levels were increased during iTreg differentiation, and related mRNA levels also was increased. CONCLUSION: IL-12A can promote the differentiation of iTreg and synergistic effect on the inhibition of effector T cells.

Key words: interleukin 12A, regulatory T cells, effect T cells, TGF-β1

CLC Number:

R965.2

ZHANG Xiaoling，GUAN Qiaobin，YANG Yi, GUO Li，HAN Chenyang. Effect of interleukin 12A on proliferation and differentiation of CD4+CD25+Foxp3+Treg[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(5): 496-502.

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