Aimulajiang Kalibixiati, Wen Zhaohai, Tian Xiaowei, Lakho Shakeel Ahmed, Zhang Yang, Naqvi Muhammad Ali-Ul-Husnain, Liang Meng, Song Xiaokai, Xu Lixin, Li Xiangrui, Yan Ruofeng
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
Animals (Basel). 2020 Nov 17;10(11):2137. doi: 10.3390/ani10112137.
Ephrin domain containing protein (EPH), a significant excreted and secreted product (ESPs) of , has been identified to have antigenic functions. Over the past years, a new subset of CD4 + T named as T helper 9 cells that secrete interleukin-9 (IL-9) as a signature cytokine is associated with tumor immunity and allergy. Nonetheless, the understanding of immunomodulatory roles of EPH on goat Th9 and other immune cells remains limited. Herein, EPH from (HcEPH) was cloned and expressed in pET-28a. Immunofluorescence assay (IFA) was carried-out to localize rHcEPH within adult worms and to bind with goat peripheral blood mononuclear cells (PBMCs). Besides, the impact of rHcEPH on signature cytokine IL-9 expression in goat PBMCs was evaluated. Flow cytometry was employed to examine Th9 cells production and cell apoptosis. The results revealed success in the expression and localization of rHcEPH in surface of adult gut sections. According to IFA analysis, the rHcEPH protein was capable to react precisely with anti- antibodies. Further functional analysis showed that correlation between rHcEPH and host PBMCs significantly enhanced Th9 cell differentiation, IL-9 expression, cell apoptosis efficiency, and cell migration, whereas cell proliferation was suppressed significantly depending on the concentration. Our observations indicated that rHcEPH protein is linked to modulate the host immune cells and could enhance protective immunity by inducing Th9 responses.
含Ephrin结构域蛋白（EPH）是[具体来源]的一种重要的排泄和分泌产物（ESPs），已被确定具有抗原功能。在过去几年中，一种新的CD4 + T细胞亚群，即分泌白细胞介素-9（IL-9）作为特征性细胞因子的辅助性T9细胞，与肿瘤免疫和过敏相关。然而，对EPH在山羊Th9细胞和其他免疫细胞上的免疫调节作用的了解仍然有限。在此，克隆了来自[具体来源]的EPH（HcEPH）并在pET-28a中表达。进行免疫荧光分析（IFA）以将重组HcEPH（rHcEPH）定位在成虫体内，并使其与山羊外周血单个核细胞（PBMCs）结合。此外，评估了rHcEPH对山羊PBMCs中特征性细胞因子IL-9表达的影响。采用流式细胞术检测Th9细胞的产生和细胞凋亡。结果显示rHcEPH成功表达并定位在成虫肠道切片表面。根据IFA分析，rHcEPH蛋白能够与抗[具体抗体]精确反应。进一步的功能分析表明，rHcEPH与宿主PBMCs之间的相关性显著增强了Th9细胞分化、IL-9表达、细胞凋亡效率和细胞迁移，而细胞增殖则根据浓度显著受到抑制。我们的观察结果表明，rHcEPH蛋白与调节宿主免疫细胞有关，并可通过诱导Th9反应增强保护性免疫。
