title: Inflammatory responses to a Clostridium perfringens type A strain and α-toxin in primary intestinal epithelial cells of chicken embryos.
abstract: The causative pathogen of necrotic enteritis is the Gram-positive bacterium Clostridium perfringens. Its main cell wall component, peptidoglycan (PGN), can be recognized by Toll-like receptor 2 and nucleotide-binding oligomerization domain (NOD). Consequently, the immune response is initiated via activation of nuclear factor kappa B (NF-κB) signalling pathway. An in vitro study was conducted to investigate chicken intestinal inflammatory responses to C. perfringens type A and one of its virulence factors, α-toxin. In primary intestinal epithelial cells, C. perfringens as well as commercially available PGN and α-toxin challenge upregulated mRNA expression of interleukin (IL)-6, IL-8 and inducible nitric oxide synthase (iNOS) with a dosage-dependent manner at 3 h post infection (p.i.; P ≤ 0.001). Time-course effects of three stimulators at high concentration were further examined. C. perfringens infection elevated IL-6, IL-8 and iNOS levels from 1 h to 9 h p.i., while PGN treatment increased IL-6 and IL-8 expression at 1 h and 3 h p.i. (P < 0.05). Bacterial and PGN treatments induced NOD1 expression at 6 h p.i. and only bacterial infection boosted NF-κB p65 expression at 6 h and 9 h p.i. (P < 0.05). α-Toxin treatment upregulated IL-6 and IL-8 expression throughout infection, as well as iNOS, TNF-α and NF-κB p65 expression at later hours p.i. (P < 0.05). In conclusion, both C. perfringens and α-toxin challenge induced intense cytokine expression associated with NF-κB activation in chicken intestinal epithelial cells. The receptors for the recognition of PGN component of C. perfringens need further investigation.
authors.affiliation: a State Key Laboratory of Animal Nutrition, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, College of Animal Science and Technology , China Agricultural University , Beijing 100193 , China.
authors.forename: Shuangshuang
authors.initials: S
authors.lastname: Guo
authors.forename: Changwu
authors.initials: C
authors.lastname: Li
authors.forename: Dan
authors.initials: D
authors.lastname: Liu
authors.forename: Yuming
authors.initials: Y
authors.lastname: Guo
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000633:toxicity*; D002135:Calcium-Binding Proteins / Q000633:toxicity*; D002642:Chick Embryo; D003016:Clostridium perfringens / Q000276:immunology*; D004847:Epithelial Cells / Q000187:drug effects* / Q000276:immunology* / Q000382:microbiology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology*; D066298:In Vitro Techniques; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D007413:Intestinal Mucosa / Q000166:cytology*; D016328:NF-kappa B / Q000276:immunology; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D015398:Signal Transduction / Q000276:immunology*; D010738:Type C Phospholipases / Q000633:toxicity*
chemical_list: D001427:Bacterial Toxins; D002135:Calcium-Binding Proteins; D015850:Interleukin-6; D016209:Interleukin-8; D016328:NF-kappa B; D052247:Nitric Oxide Synthase Type II; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens