Factor associated suicide ligand (FasL) participates in the intestinal immune response to muramyl dipeptide challenge in grass carp Ctenopharyngodon idella

Factor associated suicide ligand (FasL) is a multifunctional member of tumor necrosis factor ligand (TNF) superfamily, which exerts vital effects on maintaining homeostasis in the immune system. However, the functions of FasL in intestinal immunity of teleost fish are unknown. This study detected and characterized a fish FasL homolog (defined as CiFasL) in grass carp. The deduced CiFasL protein contained a conserved TNF homology domain (THD) and a representative transmembrane region. Expression profile analysis indicated that CiFasL was widely expressed in the tested tissues and developmental stages of grass carp, and that its mRNA level was significantly up-regulated after being challenged by Aeromonas hydrophila, A. veronii, and muramyl dipeptide (MDP) in vivo. Recombinant CisFasL (rCisFasL) was found to up-regulate pro-apoptotic genes (FasL, FADD, Caspase-8 and Caspase-3) expression in the intestine time-dependently. Moreover, rCisFasL protein effectively suppressed the expression of intestinal inflammatory cytokines (TNF-α, IL-1β, IL-6 and IL-8) and PepT1/NOD2 pathway signaling molecules (PepT1, NOD2, RIP2, p38MAPK and NF-κB) in response to MDP challenge. Finally, CiFasL silencing aggravated the MDP-mediated intestinal inflammation by inhibiting PepT1/NOD2 pathway activation in intestine of grass carp. Collectively, these findings provide the first experimental demonstration that CiFasL plays a negative regulatory role in MDP-induced intestinal inflammation.