A novel avian intestinal epithelial cell line: its characterization and exploration as an in vitro infection culture model for Eimeria species

作者：Huifang Chen, Juan Li, Xiaoting Pan, Zhichao Hu, Jianfeng Cai, Zijie Xia, Nanshan Qi, Shenquan Liao, Zachary Spritzer, Yinshan Bai, Mingfei Sun · 发表于：Parasites & Vectors · 年份：2024 · DOI：10.1186/s13071-023-06090-8 · 被引用次数：6 · 研究领域：Coccidia and coccidiosis research、Bird parasitology and diseases、Animal Nutrition and Physiology

BACKGROUND: The gastrointestinal epithelium plays an important role in directing recognition by the immune system, and epithelial cells provide the host's front line of defense against microorganisms. However, it is difficult to cultivate avian intestinal epithelial cells in vitro for lengthy periods, and the lack of available cell lines limits the research on avian intestinal diseases and nutritional regulation. Chicken coccidiosis is a serious intestinal disease that causes significant economic losses in the poultry industry. In vitro, some cell line models are beneficial for the development of Eimeria species; however, only partial reproduction can be achieved. Therefore, we sought to develop a new model with both the natural host and epithelial cell phenotypes. METHODS: In this study, we use the SV40 large T antigen (SV40T) gene to generate an immortalized cell line. Single-cell screening technology was used to sort positive cell clusters with epithelial characteristics for passage. Polymerase chain reaction (PCR) identification, immunofluorescence detection, and bulk RNA sequencing analysis and validation were used to check the expression of epithelial cell markers and characterize the avian intestinal epithelial cell line (AIEC). AIECs were infected with sporozoites, and their ability to support the in vitro endogenous development of Eimeria tenella was assessed. RESULTS: This novel AIEC consistently expressed intestinal epithelial markers. Transcriptome assays revealed...