Lack of LTβR Increases Susceptibility of IPEC-J2 Cells to Porcine Epidemic Diarrhea Virus.
Lack of LTβR Increases Susceptibility of IPEC-J2 Cells to Porcine Epidemic Diarrhea Virus.
复制标题
缺乏 LTβR 会增加 IPEC-J2 细胞对猪流行性腹泻病毒的敏感性
作者：
Altawaty T;Liu L;Zhang H;Tao C;Hou S;Li K;Wang Y
外周淋巴器官的发育和维持需要光敏素β受体（LTβR）。有证据表明LTβR参与多种细胞过程;然而，它是否在维持猪肠道肠上皮细胞（IPEC-J2）的细胞功能中发挥作用，特别是在猪流行性腹泻病毒（PEDV）感染期间，仍然未知。在这项研究中，我们使用CRISPR/Cas9产生了LTβR null IPEC-J2细胞，以检查LTβR在细胞增殖，凋亡和对PEDV感染的反应中的重要性。我们的研究结果表明，LTβR的缺乏导致细胞增殖显着下降，可能是由于LTβR−/− IPEC-J2细胞的S期阻滞。使用无标记数字全息显微镜记录两种细胞类型的三维形态长达72小时，并显示发生凋亡的LTβR−/−细胞数量显著增加。此外，我们发现PEDV感染的LTβR−/− null IPEC-J2细胞表现出对活化B细胞（NFκB）核因子κ轻链增强子靶基因（白细胞介素（IL）-6和IL-8）以及粘膜屏障完整性相关基因（血管细胞粘附分子1（VCAM 1）和IL-22）的显著抑制，这可能解释了为什么LTβR−/−细胞对PEDV感染更敏感。总的来说，我们的数据不仅证明了LTβR在猪肠上皮细胞中的关键作用，而且为更好地理解PEDV感染的细胞反应提供了数据。
The essential requirement of the lymphotoxin beta receptor (LTβR) in the development and maintenance of peripheral lymphoid organs is well recognized. Evidence shows that LTβR is involved in various cellular processes; however, whether it plays a role in maintaining the cellular function of intestinal porcine enterocytes (IPEC-J2), specifically during porcine epidemic diarrhea virus (PEDV) infection, remains unknown. In this study, we generated LTβR null IPEC-J2 cells using CRISPR/Cas9 to examine the importance of LTβR in cell proliferation, apoptosis, and the response to PEDV infection. Our results showed that the lack of LTβR leads to significantly decreased cell proliferation, potentially due to S phase arrest in LTβR−/− IPEC-J2 cells. Label-free digital holographic microscopy was used to record the three-dimensional morphology of both cell types for up to 72 hours and revealed significantly increased numbers of LTβR−/− cells undergoing apoptosis. Furthermore, we found that PEDV-infected LTβR−/− null IPEC-J2 cells exhibited significant suppression of nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) target genes (interleukin (IL)-6 and IL-8) and mucosal barrier integrity-related genes (vascular cell adhesion molecule 1 (VCAM1) and IL-22), which may explain why LTβR−/− cells are more susceptible to PEDV infection. Collectively, our data not only demonstrate the key role of LTβR in intestinal porcine enterocytes, but also provide data for the improved understanding of the cellular response to PEDV infection.
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