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Abstract：
BACKGROUND: Haemonchus contortus is a parasite widely distributed in tropical, subtropical, and warm temperate regions, causing significant economic losses in the livestock industry worldwide. However, little is known about the genetics of H. contortus resistance in livestock. In this study, we monitor the dynamic immune cell responses in diverse peripheral blood mononuclear cells (PBMCs) during H. contortus infection in goats through single-cell RNA sequencing (scRNA-Seq) analysis.
RESULTS: A total of four Boer goats, two goats with oral infection with the L3 larvae of H. contortus and two healthy goats as controls, were used in the animal test. The infection model in goats was established and validated by the fecal egg count (FEC) test and qPCR analysis of the gene expression of IL-5 and IL-6. Using scRNA-Seq, we identified seven cell types, including T cells, monocytes, natural killer cells, B cells, and dendritic cells with distinct gene expression signatures. After identifying cell subpopulations of differentially expressed genes (DEGs) in the case and control groups, we observed the upregulation of multiple inflammation-associated genes, including NFKBIA and NFKBID. Kyoto Encyclopedia of the Genome (KEGG) enrichment analysis revealed significant enrichment of NOD-like receptor pathways and Th1/Th2 cell differentiation signaling pathways in CD4 T cells DEGs. Furthermore, the analysis of ligand-receptor interaction networks showed a more active state of cellular communication in the PBMCs from the case group, and the inflammatory response associated MIF-(CD74 + CXCR4) ligand receptor complex was significantly more activated in the case group, suggesting a potential inflammatory response.
CONCLUSIONS: Our study preliminarily revealed transcriptomic profiling characterizing the cell type specific mechanisms in host PBMCs at the single-cell level during H. contortus infection.
RESULTS: A total of four Boer goats, two goats with oral infection with the L3 larvae of H. contortus and two healthy goats as controls, were used in the animal test. The infection model in goats was established and validated by the fecal egg count (FEC) test and qPCR analysis of the gene expression of IL-5 and IL-6. Using scRNA-Seq, we identified seven cell types, including T cells, monocytes, natural killer cells, B cells, and dendritic cells with distinct gene expression signatures. After identifying cell subpopulations of differentially expressed genes (DEGs) in the case and control groups, we observed the upregulation of multiple inflammation-associated genes, including NFKBIA and NFKBID. Kyoto Encyclopedia of the Genome (KEGG) enrichment analysis revealed significant enrichment of NOD-like receptor pathways and Th1/Th2 cell differentiation signaling pathways in CD4 T cells DEGs. Furthermore, the analysis of ligand-receptor interaction networks showed a more active state of cellular communication in the PBMCs from the case group, and the inflammatory response associated MIF-(CD74 + CXCR4) ligand receptor complex was significantly more activated in the case group, suggesting a potential inflammatory response.
CONCLUSIONS: Our study preliminarily revealed transcriptomic profiling characterizing the cell type specific mechanisms in host PBMCs at the single-cell level during H. contortus infection.
摘要：
背景：扭曲的Haemonchuscontortus是一种广泛分布于热带地区的寄生虫，亚热带,和温暖的温带地区，在全世界的畜牧业中造成重大的经济损失。然而,关于牲畜中H.contrortus抗性的遗传学知之甚少。在这项研究中,我们通过单细胞RNA测序(scRNA-Seq)分析,监测山羊在扭曲嗜血杆菌感染期间不同外周血单个核细胞(PBMC)的动态免疫细胞反应.
结果：总共四只波尔山羊，两只口腔感染H.contortusL3幼虫的山羊和两只健康山羊作为对照，用于动物试验。建立山羊感染模型，并通过粪便卵计数（FEC）测试和IL-5和IL-6基因表达的qPCR分析进行验证。使用scRNA-Seq，我们确定了七种细胞类型，包括T细胞,单核细胞,自然杀伤细胞,B细胞,和具有不同基因表达特征的树突状细胞。在病例组和对照组中鉴定了差异表达基因(DEGs)的细胞亚群,我们观察到多种炎症相关基因的上调，包括NFKBIA和NFKBID。京都基因组百科全书（KEGG）富集分析揭示了CD4T细胞DEGs中NOD样受体途径和Th1/Th2细胞分化信号通路的显着富集。此外，配体-受体相互作用网络的分析显示，病例组的PBMC中细胞通讯更活跃，而炎症反应相关的MIF-(CD74+CXCR4)配体受体复合物在病例组中明显激活,提示潜在的炎症反应。
结论：我们的研究初步揭示了转录组分析特征，表明H.contortus感染期间宿主PBMC在单细胞水平上的细胞类型特异性机制。
结果：总共四只波尔山羊，两只口腔感染H.contortusL3幼虫的山羊和两只健康山羊作为对照，用于动物试验。建立山羊感染模型，并通过粪便卵计数（FEC）测试和IL-5和IL-6基因表达的qPCR分析进行验证。使用scRNA-Seq，我们确定了七种细胞类型，包括T细胞,单核细胞,自然杀伤细胞,B细胞,和具有不同基因表达特征的树突状细胞。在病例组和对照组中鉴定了差异表达基因(DEGs)的细胞亚群,我们观察到多种炎症相关基因的上调，包括NFKBIA和NFKBID。京都基因组百科全书（KEGG）富集分析揭示了CD4T细胞DEGs中NOD样受体途径和Th1/Th2细胞分化信号通路的显着富集。此外，配体-受体相互作用网络的分析显示，病例组的PBMC中细胞通讯更活跃，而炎症反应相关的MIF-(CD74+CXCR4)配体受体复合物在病例组中明显激活,提示潜在的炎症反应。
结论：我们的研究初步揭示了转录组分析特征，表明H.contortus感染期间宿主PBMC在单细胞水平上的细胞类型特异性机制。
