稳定表达人IFITM3的猪IPEC-J2细胞系的构建及其对PEDV增殖的影响

打开文本图片集

中图分类号：S852.659.6 文献标志码：A 文章编号：0366-6964（2025）10-5180-10

Abstract： Previous genome-wide CRISPR screening in human hepatocellular carcinoma cells identified human interferon-induced transmembrane protein 3 （IFITM3）as a critical host factor regulating porcine epidemic diarrhea virus （PEDV） infection. However，its functional role in porcinederived cells remains unexplored. Here，we generated a porcine IPEC-J2 celline stablely overexpressing hIFITM3 （IPEC-J2hiFim3）and systematically evaluated its impact on viral proliferation. The recombinant lentiviral vector encoding hIFITM3 was co-transfected with packaging plasmids pMD2. G/psPAX2 into HEK293T cells to produce replication-incompetent lentiviral particles. Following transduction of IPEC-J2 cells and puromycin selection， monoclonal cels were established via limiting dilution. Robust hIFITM3 overexpresson was confirmed by Western blot，indirect immunofluorescence assay （IFA），and RT-qPCR. CCK-8 assays verified no significant cytotoxicity associated with hIFITM3 expresson. Functional validation demonstrated that IPECJ2hIFITM3 cells potently suppressed vesicular stomatitis virus （VSV-GFP） replication， consistent with IFITM3’s canonical antiviral function. Strikingly，challenge with PEDV DR13-GFP revealed a paradoxical pro-viral effect： IPEC-J2hrim3 cels exhibited a1 Ooo-fold increase in viral titer （TCID50∙mL-1 ） compared to wild-type cells at 24hpi . This study establishes a porcine intestinal epithelial model stably expressing hIFITM3，which unexpectedly potentiates PEDV replication. These findings not only highlight the species-specific functional divergence of IFITM3 but also provide a unique platform for investigating PEDV-host interactions， optimizing viral cultivation，and developing targeted antiviral therapeutics.

Keywords： porcine epidemic diarrhea virus；IPEC-J2 cells;IFITM3；cell line （204号 ∗ Corresponding authors： LI Yongtao， E-mail： yongtaole@126. com；YU Linyang，E-mail：linyangyuhenau@163.com

猪流行性腹泻（porcineepidemicdiarrhea，PED）是由猪流行性腹泻病毒（porcineepidemicdi-arrheavirus，PEDV）引起的一种急性、高度接触性肠道传染病，主要侵袭新生仔猪的肠上皮细胞，可引起肠道绒毛萎缩和细胞坏死，临床表现为仔猪的呕吐、腹泻、厌食，严重时脱水死亡[]。（剩余16157字）