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转录组和蛋白质组分析揭示了捻转血矛线虫重组半乳糖凝集素下调山羊 PBMC 功能,并在体外调节几个信号通路。

Transcriptional and proteomic analysis reveal recombinant galectins of Haemonchus contortus down-regulated functions of goat PBMC and modulation of several signaling cascades in vitro.

机构信息

College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China.

College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China.

出版信息

J Proteomics. 2014 Feb 26;98:123-37. doi: 10.1016/j.jprot.2013.12.017. Epub 2014 Jan 5.

Abstract

UNLABELLED

In this study, a combined proteomic and transcriptomic analysis was performed to understand the mechanisms underlying the immunomodulation induced by recombinant galectins of Haemonchus contortus (rHco-gal-m/f) on goat peripheral blood mononuclear cells (PBMC). We demonstrated that rHco-gal-m/f could be distinguished by antisera from goats experimentally infected with H. contortus and bound to the surface of goat PBMC. Following rHco-gal-m/f exposure, 16 differentially expressed proteins were identified, which function in biological processes such as stimulus response, biological regulation and localization. According to Gene Ontology Annotation, 15 proteins (93.8%) had binding activity and 9 proteins (56.3%) had catalytic activity. A series of transcriptomic analyses were performed subsequently to assess the expression change of certain pathway members. The integrated results of proteomic and transcriptomic analysis suggested that the activation of VEGF pathway, free radical producing pathway, NFκB pathway and ubiquitin-proteasome pathway was inhibited following exposure to rHco-gal-m/f, while the TLR pathway and CASPASE pathway were activated. Cytokine production and T cell differentiation were also influenced. Cell migration assays and ELISA were performed and the results were in accordance with the change of the proteins and genes. The protein and gene profiles determined here identified several mechanisms underlying the rHco-gal-m/f-induced immunomodulation of goat PBMC.

BIOLOGICAL SIGNIFICANCE

This research provided insight into the interactive relationship between parasitic nematode galectins and host PBMC. It also shed new lights on the understanding of molecular mechanisms of helminthic immune evasion.

摘要

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在这项研究中,我们进行了一项联合蛋白质组学和转录组学分析,以了解重组 Haemonchus contortus 半乳糖凝集素(rHco-gal-m/f)对山羊外周血单核细胞(PBMC)诱导免疫调节的机制。我们证明,rHco-gal-m/f 可以被来自实验感染 Haemonchus contortus 的山羊的抗血清识别,并与山羊 PBMC 表面结合。rHco-gal-m/f 暴露后,鉴定出 16 个差异表达蛋白,这些蛋白在刺激反应、生物调节和定位等生物学过程中发挥作用。根据基因本体论注释,15 个蛋白(93.8%)具有结合活性,9 个蛋白(56.3%)具有催化活性。随后进行了一系列转录组分析,以评估某些途径成员表达变化。蛋白质组学和转录组学分析的综合结果表明,rHco-gal-m/f 暴露后,VEGF 途径、自由基产生途径、NFκB 途径和泛素-蛋白酶体途径的激活受到抑制,而 TLR 途径和 CASPASE 途径被激活。细胞因子产生和 T 细胞分化也受到影响。进行了细胞迁移测定和 ELISA,结果与蛋白质和基因的变化一致。这里确定的蛋白质和基因图谱确定了 rHco-gal-m/f 诱导山羊 PBMC 免疫调节的几种机制。

生物学意义

这项研究深入了解了寄生线虫半乳糖凝集素与宿主 PBMC 之间的相互关系。它还为理解寄生虫免疫逃避的分子机制提供了新的思路。

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