Trichinella spiralis galectin binding to toll-like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosa

Previous studies showed that Trichinella spiralis galectin (Tsgal) facilitates larval invasion of intestinal epithelium cells (IECs). However, IEC proteins binding with Tsgal were not identified, and the mechanism by which Tsgal promotes larval invasion is not clear. Toll-like receptors (TLRs) are protein receptors responsible for recognition of pathogens. The aim of this study was to investigate whether recombinant Tsgal (rTsgal) binds to TLR-4, activates inflammatory pathway in gut epithelium and mediates T. spiralis invasion. Indirect immunofluorescence (IIF), GST pull-down and co-immunoprecipitation (Co-IP) assays confirmed specific binding between rTsgal and TLR-4 in Caco-2 cells. qPCR and Western blotting showed that binding of rTsgal with TLR-4 up-regulated the TLR-4 transcription and expression in Caco-2 cells, and activated p-NF-kappa B p65 and p-ERK1/2. Activation of inflammatory pathway TLR-4/MAPK-NF-kappa B by rTsgal up-regulated pro-inflammatory cytokines (IL-1 beta and IL-6) and down-regulated anti-inflammatory cytokine TGF-beta in Caco-2 cells, and induced intestinal inflammation. TAK-242 (TLR-4 inhibitor) and PDTC (NF-kappa B inhibitor) significantly inhibited the activation of TLR-4 and MAPK-NF-kappa B pathway. Moreover, the two inhibitors also inhibited IL-1 beta and IL-6 expression, and increased TGF-beta expression in Caco-2 cells. In T. spiralis infected mice, the two inhibitors also inhibited the activation of TLR-4/MAPK-NF-kappa B pathway, ameliorated intestinal inflammation, impeded larval invasion of gut mucosa and reduced intestinal adult burdens. The results showed that rTsgal binding to TLR-4 in gut epithelium activated MAPK-NF-kappa B signaling pathway, induced the expression of TLR-4 and pro-inflammatory cytokines, and mediated larval invasion. Tsgal might be regarded as a candidate molecular target of vaccine against T. spiralis enteral invasive stage.

Automated summary

This study demonstrates that Tsgal protein binds to TLR-4 and activates an inflammatory pathway, mediating the invasion of T. spiralis larvae in the gut. These findings suggest that Tsgal could be a potential molecular target for a vaccine against the enteral invasive stage of T. spiralis.