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重组捻转血矛线虫电子传递黄素蛋白α亚基蛋白对体外山羊免疫细胞的调节功能。

Modulatory functions of recombinant electron transfer flavoprotein α subunit protein from Haemonchus contortus on goat immune cells in vitro.

机构信息

MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China; School of Life Science, Huizhou University, Huizhou 516007, PR China.

MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China.

出版信息

Vet Parasitol. 2020 Dec;288:109300. doi: 10.1016/j.vetpar.2020.109300. Epub 2020 Nov 1.

Abstract

Suppression and modulation of the host immune response to parasitic nematodes have been extensively studied. In the present study, we cloned and produced recombinant electron transfer flavoprotein α subunit (ETFα) protein from Haemonchus contortus (rHCETFα), a parasitic nematode of small ruminants, and studied the effect of this protein on modulating the immune response of goat peripheral blood mononuclear cells (PBMCs) and monocytes. Immunohistochemical tests verified that the HCETFα protein was localized mainly in the intestinal wall and on the body surface of worms. Immunoblot analysis revealed that rHCETFα was recognized by the serum of goats artificially infected with H. contortus. Immunofluorescence analysis indicated that rHCETFα bound to the surface of PBMCs. rHCETFα was co-incubated with goat PBMCs to observe the immunomodulatory effects exerted by HCETFα on proliferation, apoptosis, cytokine secretion and nitric oxide (NO) production. The results showed that rHCETFα suppressed the proliferation of goat PBMCs stimulated by concanavalin A and induced apoptosis in goat PBMCs. After rHCETFα exposure, IL-2, IL-4, IL-17A and TNF-α expression was markedly reduced, whereas secretion of TGF-β1 was significantly elevated, in goat PBMCs. Moreover, rHCETFα up-regulated NO production in a dose-dependent manner. FITC-dextran internalization assays showed that rHCETFα inhibited phagocytosis of goat monocytes. These results elucidate the interaction between parasites and hosts at the molecular level, suggest a possible immunomodulatory target and contribute to the search for innovative proteins that may be candidate targets for drugs and vaccines.

摘要

宿主对寄生线虫免疫反应的抑制和调节已经得到了广泛的研究。在本研究中,我们从反刍动物寄生线虫捻转血矛线虫中克隆并表达了重组电子传递黄素蛋白α亚基(rHCETFα)蛋白,并研究了该蛋白对调节山羊外周血单核细胞(PBMC)和单核细胞免疫反应的影响。免疫组织化学试验证实,HCETFα蛋白主要定位于虫体的肠壁和体表。免疫印迹分析显示,rHCETFα可被人工感染捻转血矛线虫的山羊血清识别。免疫荧光分析表明,rHCETFα与 PBMC 的表面结合。将 rHCETFα与山羊 PBMC 共孵育,观察 HCETFα对 PBMC 增殖、凋亡、细胞因子分泌和一氧化氮(NO)产生的免疫调节作用。结果表明,rHCETFα抑制了刀豆蛋白 A 刺激的山羊 PBMC 的增殖,并诱导了山羊 PBMC 的凋亡。rHCETFα暴露后,山羊 PBMC 中 IL-2、IL-4、IL-17A 和 TNF-α的表达明显降低,而 TGF-β1 的分泌显著升高。此外,rHCETFα呈剂量依赖性地上调了 NO 的产生。FITC-葡聚糖摄取试验表明,rHCETFα抑制了山羊单核细胞的吞噬作用。这些结果从分子水平阐明了寄生虫和宿主之间的相互作用,提示了一种可能的免疫调节靶点,并有助于寻找可能成为药物和疫苗候选靶点的创新蛋白。

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