Abstract
BackgroundToxoplasma gondii can invade and replicate in all nucleated cells in a wide range of host species, and infection induces IL-1β production. IL-1β plays central roles in the stimulation of the innate immune system and inflammation. However, little is known of the innate immune responses in human fetal small intestinal epithelial cells (FHs 74 Int cells) after T. gondii infection.MethodsFHs 74 Int cells were infected with the T. gondii GFP-RH strain. Then, IL-1β production and its mechanisms of action were evaluated using ELISA, MTT cell viability assays, Western blotting, immunofluorescence, quantitative real-time polymerase chain reaction (qRT-PCR), and gene-specific small interfering RNA (siRNA) transfection.ResultsInfection of FHs 74 Int cells by T. gondii triggered significant time- and dose-dependent IL-1β production. Although T. gondii activated NLRP1, NLRP3, NLRC4 and AIM2 inflammasomes in FHs 74 Int cells, NLRP3 levels were consistently and significantly time-dependently increased, while the other inflammasomes were not. Transfection with siRNA targeting NLRP3, cleaved caspase-1 (Casp-1) or ASC significantly reduced T. gondii-induced IL-1β production, whereas T. gondii proliferation was markedly increased. Toxoplasma gondii infection activated P2X7 receptor (P2X7R) levels in FHs 74 Int cells in a time-dependent manner; however, transfection with siRNA targeting P2X7R significantly reduced T. gondii-induced IL-1β secretion and substantially increased T. gondii proliferation, which is mediated by decreased protein expression levels of NLRP3, cleaved Casp-1 and ASC. Collectively, NLRP3-dependent IL-1β secretion is mediated by P2X7R in small intestinal epithelial cells in response to T. gondii infection, thereby controlling parasite proliferation.ConclusionsThis study revealed that the P2X7R/NLRP3 pathway plays important roles in IL-1β secretion and inhibition of T. gondii proliferation in small intestinal epithelial cells. These results not only contribute to our understanding of the mucosal immune mechanisms of T. gondii infection but also offer new insight into the identification of innate resistance in the gut epithelium.
Highlights
Toxoplasma gondii can invade and replicate in all nucleated cells in a wide range of host species, and infection induces IL-1β production
Toxoplasma gondii induces IL-1β secretion in small intestinal epithelial cells To evaluate the capacity of small intestinal epithelial cells to produce IL-1β in response to T. gondii, FHs 74 Int cells were infected with live T. gondii at different multiplicity of infection (MOI) for 8 h, and IL-1β levels in the cell culture supernatants were measured by Enzyme-linked immunosorbent assay (ELISA)
Our findings reveal the role of P2XR in the induction of NLRP3 inflammasome activation in T. gondii-induced small intestinal epithelial cells
Summary
Toxoplasma gondii can invade and replicate in all nucleated cells in a wide range of host species, and infection induces IL-1β production. Little is known of the innate immune responses in human fetal small intestinal epithelial cells (FHs 74 Int cells) after T. gondii infection. Toxoplasma gondii is an obligate intracellular protozoan parasite that can invade and replicate in all nucleated cells. It is prevalent in humans and animals worldwide, and one-third of the world’s population is reportedly infected with T. gondii [1]. A single infected cat can shed millions of oocysts into the environment These cysts can be taken up by intermediate hosts, which include virtually any warm-blooded animal, including humans. The exact roles of the small intestinal epithelium in the activation of innate immunity against T. gondii infection remain poorly understood
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