Recombinant interleukin-4 enhances Campylobacter jejuni invasion of intestinal pig epithelial cells (IPEC-1)

Campylobacter jejuni, a leading cause of bacterial gastroenteritis, has a diverse spectrum of disease expression. Polymicrobial infections may contribute to this, such as Trichuris, which elicits type 2 cytokines (including IL-4) and downregulates type 1 immunity. In previous studies, gnotobiotic piglets infected with C. jejuni and Trichuris suis had bloody diarrhea and marked gastrointestinal pathology, including bacterial invasion into epithelial cells and macrophages. Neonatal swine given these dual infections had elevated IL-4 and IL-10 responses in feces. In the studies reported here, we hypothesized that IL-4 or IL-10 enhances invasion of intestinal pig epithelial cells (IPEC-1) by C. jejuni. 10-14-day-old IPEC-1 cells were pretreated with recombinant IL-4 (rIL-4) or rIL-10 for 5h and then challenged with C. jejuni. Cells pretreated with rIL-4 were viable and showed approximately 6-fold increases in C. jejuni (but not Escherichia coli DH5alpha) internalization compared to cells with no pretreatment. Enhanced C. jejuni invasion was rIL-4 dose-dependent and reversed by addition of anti-IL-4 antibody. Preincubation with rIL-10 did not significantly alter C. jejuni internalization. Transepithelial electrical resistance (TEER) was significantly reduced following rIL-4 treatment, but not rIL-10 treatment. After rIL-4 pretreatment and C. jejuni challenge, light microscopy showed vacuolated cells with damaged paracellular junctions. Transmission electron microscopy (TEM) showed multiple internalized bacteria. Most were in the cytoplasm, but some were within or adjacent to vacuoles. We conclude that rIL-4 damages paracellular junctions and alters the physiology of these epithelial cells allowing increased invasion of C. jejuni.