绵羊痒螨巨噬细胞迁移抑制因子重组蛋白对兔外周血单个核细胞Th1/Th2型和Th17/Treg型特征因子表达的影响

doi:10.11843/j.issn.0366-6964.2020.03.019

Abstract

Abstract: To explore the effects of macrophage migration inhibitory factor of Psoroptes ovis (PoMIF) on Th1/Th2 and Th17/Treg balance of rabbit peripheral blood mononuclear cells (PBMC) in vitro, the PoMIF gene was amplified from the extracted total RNA from P. ovis mites by the reverse transcription-polymerase chain reaction (RT-PCR), and the recombinant protein rPoMIF was expressed with prokaryotic expression system, then the oxidoreductase and tauromerase activities of rPoMIF was evaluated. PBMCs isolated from healthy New Zealand rabbits were cultured with different concentrations of rPoMIF, and the optimum concentration of rPoMIF was screened. Finally, the expression levels of Th1, Th2, Th17 and Treg-specific transcription factors of T-bet, GATA-3, RORc, Foxp3 and specific cytokines of IFN-γ, IL-4, IL-17A and IL-10 in PBMC incubated with the optimal concentration of rPoMIF for 0, 1, 6, 12, 24 and 36 h were detected by real time fluorescence quantitative PCR (qPCR). The results showed that the full-length of PoMIF was 363 bp, containing 357 bp open reading frame; rPoMIF was mainly in the supernatant of Escherichia coli (E. coli) lysate, with a molecular size of 32 kD (including an extra 19 kD for the attached His-tag fusion peptide); and it has tautomerism activity; the mRNA levels of Th1-specific T-bet and IFN-γ decreased firstly and then increased after treatment, while Th2-specific GATA-3 and IL-4 decreased at all tested time points, and the ratio of T-bet/GATA-3 and IFN-γ/IL-4 increased at 12, 24 and 36 h; the mRNA levels of Th17-specific RORc and IL-17A decreased at all tested time points, while Foxp3 and IL-10 from Treg cells increased at all tested time points, and the ratio of RORc/Foxp3 and IL-17A/IL-10 decreased at all tested time points. In conclusion, rPoMIF can cause the shifts of Th1/Th2 and Th17/Treg towards Th1 and Treg in PBMC of rabbits.

Key words: Psoroptes ovis, rabbit, macrophage migration inhibitory factor, peripheral blood mononuclear cells, Th1/Th2 cell balance, Th17/Treg cell balance

CLC Number:

S852.746

ZHENG Youle, CHEN Yuhang, XIE Yue, YANG Guangyou, HE Ran, GU Xiaobin. The Effects of Recombinant Macrophage Migration Inhibitory Factor of Psoroptes ovis on the Characteristic Molecules of Th1/Th2 and Th17/Treg Type Immune Responses in Rabbits Peripheral Blood Mononuclear Cells in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(3): 594-601.

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The Effects of Recombinant Macrophage Migration Inhibitory Factor of Psoroptes ovis on the Characteristic Molecules of Th1/Th2 and Th17/Treg Type Immune Responses in Rabbits Peripheral Blood Mononuclear Cells in vitro

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