Department of Microbiology, National Veterinary Institute, SE-751 89Uppsala, Sweden.
Department of Cell and Molecular Biology, BMC, Uppsala University, Box 596, SE-751 24Uppsala, Sweden.
Parasitology. 2021 May;148(6):712-725. doi: 10.1017/S0031182021000111. Epub 2021 Feb 4.
The study aimed to monitor parasite and host gene expression during the early stages of Eimeria tenella infection of chicken cells using dual RNA-Seq analysis. For this, we used chicken macrophage-like cell line HD11 cultures infected in vitro with purified E. tenella sporozoites. Cultures were harvested between 2 and 72 h post-infection and mRNA was extracted and sequenced. Dual RNA-Seq analysis showed clear patterns of altered expression for both parasite and host genes during infection. For example, genes in the chicken immune system showed upregulation early (2–4 h), a strong downregulation of genes across the immune system at 24 h and a repetition of early patterns at 72 h, indicating that invasion by a second generation of parasites was occurring. The observed downregulation may be due to immune self-regulation or to immune evasive mechanisms exerted by E. tenella. Results also suggested pathogen recognition receptors involved in E. tenella innate recognition, MRC2, TLR15 and NLRC5 and showed distinct chemokine and cytokine induction patterns. Moreover, the expression of several functional categories of Eimeria genes, such as rhoptry kinase genes and microneme genes, were also examined, showing distinctive differences which were expressed in sporozoites and merozoites.
本研究旨在使用双 RNA-Seq 分析监测鸡细胞感染柔嫩艾美耳球虫(Eimeria tenella)早期阶段的寄生虫和宿主基因表达。为此,我们使用鸡巨噬细胞样细胞系 HD11 进行体外感染纯化的柔嫩艾美耳球虫孢子进行培养。在感染后 2 至 72 小时收获培养物,并提取和测序 mRNA。双 RNA-Seq 分析显示,在感染过程中,寄生虫和宿主基因的表达均呈现明显的变化模式。例如,鸡免疫系统的基因在早期(2-4 小时)上调,24 小时时免疫系统的基因强烈下调,72 小时时又重复早期模式,表明正在发生第二代寄生虫的入侵。观察到的下调可能是由于免疫自身调节或柔嫩艾美耳球虫的免疫逃避机制所致。研究结果还表明,参与柔嫩艾美耳球虫先天识别的病原体识别受体 MRC2、TLR15 和 NLRC5 以及表现出独特趋化因子和细胞因子诱导模式的趋化因子和细胞因子诱导模式。此外,还检查了几种艾美耳属基因的功能类别,如棒状体激酶基因和微线体基因的表达,在孢子和裂殖子中表现出明显的差异。