Chicken intestine defensins activated murine peripheral blood mononuclear cells through the TLR4-NF-κB pathway

Defensins serve as alarm signals in mobilizing the immune system and activating the innate and adaptive immune responses. In order to investigate whether avian defensins could activate monocytes of another species, and whether chicken defensins could modulate or amplify the adaptive immune responses of murine through the TLR-NF-κB pathway, the relationship between the chicken intestinal defensin AvBD13 and TLR4 in murine peripheral blood mononuclear cells (PBMCs) was explored in vitro. Monocytes were stimulated with AvBD13 (1 μg/mL). The levels of NF-κB p65, CD80, CD86, IL-12 and IFN-α were measured by immunohistochemical analysis of the cells or enzyme-linked immunosorbent assay (ELISA), and the TLR4 levels in monocytes were measured by flow-cytometry. We found that AvBD13 can activate NF-κB, induce the inflammatory cytokines IL-12 and IFN-α, and upregulate costimulatory molecules like CD80 and monocyte proliferation, which was clearly inhibited by the anti-TLR4 antibody. TLR4 expression was rapidly downregulated in the presence of AvBD13. AvBD13 could modulate monocytes directly and serve as an endogenous ligand for TLR4 and upregulate costimulatory molecules and monocyte proliferation. Thus, TLR4 is involved in AvBD13-mediated activation of adaptive immune responses.