新生小牛感染性腹泻的主要原因之一是根尖丛原生动物隐孢子虫(C.parvum)。然而,对其免疫发病机制知之甚少。使用下一代测序,这项研究调查了新生小牛对C.parvum感染的免疫转录反应。新生雄性荷斯坦-弗里斯小牛口服感染(N=5)或不口服感染(CTRL组,N=5)在生命的第1天与小梭菌卵囊(gp60亚型IIaA15G2R1)一起,并在感染后第7天屠宰。从空肠粘膜提取总RNA用于短读数。使用DESeq2以<0.05的错误发现率评估感染组和CTRL组之间的差异表达基因(DEGs)。感染不影响血浆免疫血液学参数,包括中性粒细胞,淋巴细胞,单核细胞,白细胞,血小板,感染后第7天的红细胞计数以及血细胞比容和血红蛋白浓度。根据UniProt免疫系统过程数据库选择免疫相关的DEGs,并使用Cytoscape(v3.9.1)进行基因本体论(GO)和途径富集分析。基于GO分析,DEGs注释为粘膜免疫,识别和呈递抗原,嗜中性粒细胞的趋化性,嗜酸性粒细胞,自然杀伤细胞,B和T细胞介导的信号通路,包括toll样受体,白细胞介素,肿瘤坏死因子,T细胞受体,和NF-KB上调,而巨噬细胞趋化性和胞浆模式识别的标记物下调。这项研究提供了小牛C.parvum诱导的免疫相关途径的整体快照,包括新的和详细的反馈和前馈调节机制建立先天和适应性免疫应答之间的串扰在新生小牛,这可以进一步用于开发新的治疗策略。
One of the leading causes of infectious diarrhea in newborn calves is the apicomplexan protozoan Cryptosporidium parvum (C. parvum). However, little is known about its immunopathogenesis. Using next generation sequencing, this study investigated the immune transcriptional response to C. parvum infection in neonatal calves. Neonatal male Holstein-Friesian calves were either orally infected (N = 5) or not (CTRL group, N = 5) with C. parvum oocysts (gp60 subtype IIaA15G2R1) at day 1 of life and slaughtered on day 7 after infection. Total RNA was extracted from the jejunal mucosa for short read. Differentially expressed genes (DEGs) between infected and CTRL groups were assessed using DESeq2 at a false discovery rate < 0.05. Infection did not affect plasma immunohematological parameters, including neutrophil, lymphocyte, monocyte, leucocyte, thrombocyte, and erythrocyte counts as well as hematocrit and hemoglobin concentration on day 7 post infection. The immune-related DEGs were selected according to the UniProt immune system process database and were used for gene ontology (GO) and pathway enrichment analysis using Cytoscape (v3.9.1). Based on GO analysis, DEGs annotated to mucosal immunity, recognizing and presenting antigens, chemotaxis of neutrophils, eosinophils, natural killer cells, B and T cells mediated by signaling pathways including toll like receptors, interleukins, tumor necrosis factor, T cell receptor, and NF-KB were upregulated, while markers of macrophages chemotaxis and cytosolic pattern recognition were downregulated. This study provides a holistic snapshot of immune-related pathways induced by C. parvum in calves, including novel and detailed feedback and feedforward regulatory mechanisms establishing the crosstalk between innate and adaptive immune response in neonate calves, which could be utilized further to develop new therapeutic strategies.