Lack of LT?R Increases Susceptibility of IPEC-J2 Cells to Porcine Epidemic Diarrhea Virus.

ABSTRACT: The essential requirement of the lymphotoxin beta receptor (LT?R) in the development and maintenance of peripheral lymphoid organs is well recognized. Evidence shows that LT?R 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 LT?R null IPEC-J2 cells using CRISPR/Cas9 to examine the importance of LT?R in cell proliferation, apoptosis, and the response to PEDV infection. Our results showed that the lack of LT?R leads to significantly decreased cell proliferation, potentially due to S phase arrest in LT?R^-/- 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 LT?R^-/- cells undergoing apoptosis. Furthermore, we found that PEDV-infected LT?R^-/- null IPEC-J2 cells exhibited significant suppression of nuclear factor kappa-light-chain-enhancer of activated B cells (NF?B) 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 LT?R^-/- cells are more susceptible to PEDV infection. Collectively, our data not only demonstrate the key role of LT?R in intestinal porcine enterocytes, but also provide data for the improved understanding of the cellular response to PEDV infection.

SUBMITTER: Altawaty T 

PROVIDER: S-EPMC6262443 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature