Porcine Small Intestinal Epithelial Cell Line (IPEC-J2) of Rotavirus Infection As a New Model for the Study of Innate Immune Responses to Rotaviruses and Probiotics

Abstract

Previous studies of epithelial immune responses to rotavirus infection have been conducted in transformed cell lines. In this study, we evaluated a non-transformed porcine jejunum epithelial cell line (IPEC-J2) as an in-vitro model of rotavirus infection and probiotic treatment. Cell-culture-adapted porcine rotavirus (PRV) OSU strain, or human rotavirus (HRV) Wa strain, along with Lactobacillus acidophilus (LA) or Lactobacillus rhamnosus GG (LGG) were used to inoculate IPEC-J2 cells. LA or LGG treatment was applied pre- or post-rotavirus infection. We demonstrated that IPEC-J2 cells were productively infected by PRV. LA or LGG treatment of the cells did not reduce virus replication. PRV infection increased MUC3 mucin secretion. LGG treatment post-rotavirus infection reduced the mucin secretion response induced by PRV; LGG alone increased the production of membrane-associated MUC3 mucin. LA treatment prior to rotavirus infection significantly increased PRV replication and the IL-6 response to PRV infection, which is consistent with the adjuvant effect of LA. LGG treatment post-rotavirus infection downregulated the IL-6 response, confirming the anti-inflammatory effect of LGG. IPEC-J2 cells expressed toll-like receptor (TLR) 2, TLR3, and TLR9 constitutively. TLR2 expression was upregulated by LGG and peptidoglycan, corresponding to the decreased IL-6 response, indicating that the protective effect of LGG is associated with upregulation of TLR2 expression on intestinal epithelial cells. The IPEC-J2 cell model of PRV infection is a completely homologous system. It is a valuable model for studying the interactions among rotavirus-host-probiotics, and the mechanisms behind the immunomodulating effect of probiotic bacteria on innate immune responses.

Dietary Rice Bran Protects against Rotavirus Diarrhea and Promotes Th1-Type Immune Responses to Human Rotavirus Vaccine in Gnotobiotic Pigs

“…The supernatants were then divided into aliquots and stored at Ϫ20°C. Virus antigens in the fecal swabs were detected by enzyme-linked immunosorbent assay (ELISA) (18). Rectal swabs from mock treatment Gn pigs were used as negative controls.…”

Optimizing culture conditions of a porcine epithelial cell line IPEC-J2 through a histological and physiological characterization

“…Recently, a cell line from jejunum epithelium isolated from a neonatal unsuckled piglet, small intestinal porcine epithelial cell line (IPEC-J2; Berschneider 1989) was characterized and used as an in vitro model system for studying porcine intestinal pathogen-host interactions, porcine-specific pathogenesis, and innate immune responses (Schierack et al 2006;Botic et al 2007;Brown and Price 2007;Skjolaas et al 2007;Koh et al 2008;Arce et al 2010;Liu et al 2010). This cell line, a non-transformed, nontumorigenic small intestinal cell line, can secrete mucin, produce cytokines/chemokines, and express Toll-like receptors similar to those of the original tissue.…”

Involvement of PKA signalling in anti‐inflammatory effects of chitosan oligosaccharides in IPEC‐J2 porcine epithelial cells

“…IPEC-J2 cells, a cell line from jejunum epithelium isolated from a neonatal, unsuckled piglet (Berschneider, 1989), is an optimal model to study immune response in vitro. Previous studies have shown that inflammatory cytokines could be upregulated in IPEC-J2 cells exposed to LPS (Arce, Ramirez-Boo, Lucena, & Garrido, 2010), pathogenic bacteria (Geens & Niewold, 2010) and virus (Liu, Li, Wen, et al, 2010).…”