捻转血矛线虫重组磷脂酰肌醇转移蛋白对山羊外周血单个核细胞模式识别受体和细胞因子转录水平的影响

doi:10.11843/j.issn.0366-6964.2023.01.023

摘要/Abstract

摘要： 本文旨在研究捻转血矛线虫(Haemonchus contortus)磷脂酰肌醇转移蛋白(phosphatidylinositol transfer protein,HCPITP)对山羊的免疫调节功能。构建了原核表达重组质粒pET-28a-HCPITP并获得重组蛋白；分析HCPITP基因在虫卵、第三期幼虫、雌虫和雄虫中转录水平的变化；将重组蛋白与山羊外周血单个核细胞(peripheral blood mononuclear cells，PBMCs)共孵育，研究重组蛋白对山羊PBMCs增殖、模式识别受体(pattern recognition receptors,PRRs)表达和细胞因子分泌的影响。结果显示，HCPITP基因在雌虫、雄虫、第三期幼虫和虫卵中相对转录水平分别为1、10.2、0.47和0.58。Western blot分析发现该重组蛋白具有较好的反应原性。该重组蛋白能够显著抑制ConA对PBMCs的增殖作用，重组HCPITP显著促进山羊PBMCs的IL-2、IL-4、IL-6和IL-17转录水平，主要促进模式识别受体TLR1与TLR10的转录。综上，HCPITP基因转录水平具有明显的阶段差异性，重组蛋白对山羊PBMCs增殖、PRRs以及细胞因子mRNA转录水平有一定的影响作用。

关键词: 捻转血矛线虫, 磷脂酰肌醇转移蛋白, 模式识别受体, 免疫调节

Abstract: To study the effects of recombinant phosphatidylinositol transfer protein (HCPITP) from Haemonchus contortus on goat immunity, prokaryotic expression recombinant plasmid pET-28a-HCPITP was constructed and the recombinant protein was obtained. The differences of transcription level of HCPITP gene in egg, the third-stage larvae, female and male adult worm were analyzed. Goat peripheral blood mononuclear cells (PBMCs) were isolated and incubated with recombinant HCPITP, the effects of recombinant protein on the proliferation of goat PBMCs, the effects of recombinant protein on the transcription levels of pattern recognition receptors (PRRs) and cytokines in goat PBMCs were analyzed. Results showed that the relative transcription levels of HCPITP mRNA in female, male, the third-stage larvae, and egg were 1, 10.2, 0.47 and 0.58, respectively. The recombinant protein showed reactogenicity by Western blot analysis. It was found that recombinant HCPITP significantly inhibited the proliferation of ConA on PBMCs. Recombinant HCPITP promoted the transcription of IL-2, IL-4, IL-6, IL-17, TLR1 and TLR10 in goat PBMCs. In conclusion, the transcription level of HCPITP gene shows obvious stage differences, recombinant HCPITP has a certain effect on the proliferation and mRNA transcription of PRRs and cytokines of goat PBMCs.

Key words: Haemonchus contortus, phosphatidylinositol transfer protein, pattern recognition receptors, immunomodulation

中图分类号:

S852.73

谢欣然, 张玥, 陆明敏, 徐立新, 宋小凯, 李祥瑞, 严若峰. 捻转血矛线虫重组磷脂酰肌醇转移蛋白对山羊外周血单个核细胞模式识别受体和细胞因子转录水平的影响[J]. 畜牧兽医学报, 2023, 54(1): 252-262.

XIE Xinran, ZHANG Yue, LU Mingmin, XU Lixin, SONG Xiaokai, LI Xiangrui, YAN Ruofeng. Effects of Recombinant Phosphatidylinositol Transfer Protein from Haemonchus contortus on the Transcription of Pattern Recognition Receptors and Cytokines in Goat Peripheral Blood Mononuclear Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 252-262.

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