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从捻转血矛线虫幼虫中排泄和分泌的产物(72/60 kDa)在体外诱导外周血单个核细胞增殖,并激活细胞因子和 FCεR1A 受体的表达。

Excreted and secreted products (72/60 kDa) from Haemonchus placei larvae induce in vitro peripheral blood mononuclear cell proliferation and activate the expression of cytokines and FCεR1A receptor.

机构信息

Centro de Investigación Sobre Enfermedades Infecciosas, Instituto Nacional de Salud Pública, Av. Universidad No. 655, Col. Santa María Ahuacatitlán, Cuernavaca, Mor, C.P. 62100, Mexico.

Centro Nacional de Investigación Disciplinaria en Salud Animal e Inocuidad, INIFAP, Carr. Fed. Cuernavaca-Cuautla No. 8534, Col. Progreso, Jiutepec, Mor, C.P. 62550, Mexico.

出版信息

Exp Parasitol. 2019 Nov;206:107755. doi: 10.1016/j.exppara.2019.107755. Epub 2019 Sep 4.

Abstract

The aim of the present study was to assess the expression of cytokines and FCεR1A receptor stimulated by Haemonchus placei larval excretory and secretory (ES) products associated with the pathogenesis in calves. Bovine peripheral blood mononuclear cells (PBMC) were stimulated in in vitro assays with H. placei L ES product at 8, 12, 16 and 24 h. ES products were collected in in vitro assays at 48 h with molecular weight of 72/60 kDa and isoelectric point of 7.2 pI. Specific IgG for infected and control calves, positive and negative, were employed to recognise H. placei larval ES products by indirect ELISA, showing a mean of 1.8, 0.83 and 0.28 OD, respectively, (p ≤ 0.001). The quantification of relative gene expression was performed using a set of cytokines (IL-2, IFNγ, TGFβ, IL-4, IL-5, IL-6, IL-8, IL-10 and IL-13), FCεR1A receptor and housekeeping (GAPDH, β-actin and β-2-microglobulin) by RT-qPCR. An early increased expression, 2.2- to 3.4-fold change, of IL-2 (p ≤ 0.001), IL-5 and TGFβ (p ≥ 0.05) was determined, followed by TGFβ (30.7 and 14.14), IL-8 (102.8 and 1504.4) and IL-10 (60.4 and 1.7) (p ≤ 0.05) after 12 and 16 h, respectively, and reducing the expression level at 24 h. In addition, IL-6, IL-13 and FCεR1A receptor also displayed mild expression level, 2.1 - to 7.60-fold change, at 24 h (p ≥ 0.05). We conclude that ES products of 72/60 kDa collected in vitro from H. placei larvae are recognised by infected hosts and have the ability to induce diverse immune factors to modulate the nematode damage.

摘要

本研究旨在评估 Haemonchus placei 幼虫排泄分泌产物(ES)刺激细胞因子和 FCεR1A 受体的表达与小牛发病机制的关系。在体外实验中,用 H. placei L ES 产物刺激牛外周血单核细胞(PBMC),刺激时间分别为 8、12、16 和 24 小时。48 小时时在体外实验中收集分子量为 72/60 kDa 和等电点为 7.2 pI 的 ES 产物。用间接 ELISA 检测感染和对照小牛的特异性 IgG,结果分别为 1.8、0.83 和 0.28 OD(p≤0.001)。采用一套细胞因子(IL-2、IFNγ、TGFβ、IL-4、IL-5、IL-6、IL-8、IL-10 和 IL-13)、FCεR1A 受体和管家基因(GAPDH、β-肌动蛋白和β-2-微球蛋白)的 RT-qPCR 方法进行相对基因表达的定量。结果显示,IL-2(p≤0.001)、IL-5 和 TGFβ(p≥0.05)的表达水平早期升高 2.2-3.4 倍,随后 TGFβ(30.7 和 14.14)、IL-8(102.8 和 1504.4)和 IL-10(60.4 和 1.7)在 12 和 16 小时分别升高 30.7 和 14.14 倍,在 24 小时降低表达水平(p≤0.05)。此外,IL-6、IL-13 和 FCεR1A 受体在 24 小时也表现出轻微的表达水平,2.1-7.60 倍变化(p≥0.05)。综上所述,体外收集的 72/60 kDa H. placei 幼虫 ES 产物被感染宿主识别,并具有诱导多种免疫因子的能力,从而调节线虫损伤。

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