title: Role of nuclear factor-kappa B in the regulation of intercellular adhesion molecule 1 after infection of human bronchial epithelial cells by Bordetella pertussis.
abstract: Previous work has demonstrated that infection of human bronchial epithelial cells by Bordetella pertussis up-regulates intercellular adhesion molecule-1 (ICAM-1) gene and protein expression. It has also been shown that interaction of the Arg-Gly-Asp (RGD) site of filamentous hemagglutinin (FHA) with host cell very late antigen (VLA)-5 (alpha 5 beta 1 integrin) is required for the up-regulation of epithelial ICAM-1 expression, and that pertussis toxin (PT) impairs this response. We therefore examined the molecular mechanisms leading to B. pertussis-induced ICAM-1 up-regulation in BEAS-2B human bronchial epithelial cells. A colorimetric nuclear factor kappa B (NF-kappa B) activation assay demonstrated that NF-kappa B was activated in response to infection of these cells with B. pertussis. This activation occurred in an FHA(RGD)-dependent manner, and was blocked by an antibody against VLA-5, implying that binding of the RGD to VLA-5 integrin is involved in NF-kappa B activation. Western blot analysis revealed that the activation of NF-kappa B by B. pertussis was preceded by degradation of I kappa B alpha, a major cytoplasmic inhibitor of NF-kappa B. Pretreatment of the BEAS-2B cells with the NF-kappa B inhibitors pyrrolidine dithiocarbamate (PDTC), MG-132, and SN50 resulted in a marked decrease in B. pertussis-induced ICAM-1 expression, implying the involvement of NF-kappa B in ICAM-1 expression. Purified PT abrogated both NF-kappa B activation and I kappa B alpha degradation. These results suggest that ligation of VLA-5 integrin by FHA induces RGD-dependent NF-kappa B activation, thus leading to the up-regulation of epithelial ICAM-1 expression, and that a PT-sensitive G protein may be involved in this signaling pathway.
authors.affiliation: Department of Immunobiology, Meiji Pharmaceutical University, Noshio, Kiyose, Tokyo 204-8588, Japan. yishibas@my-pharm.ac.jp
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Ishibashi
authors.forename: Akemi
authors.initials: A
authors.lastname: Nishikawa
mesh_terms: D018829:Adhesins, Bacterial / Q000302:isolation & purification / Q000378:metabolism; D001886:Bordetella pertussis / Q000472:pathogenicity*; D001980:Bronchi / Q000166:cytology / Q000378:metabolism / Q000382:microbiology*; D002448:Cell Adhesion; D002457:Cell Extracts; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000032:analysis; D004847:Epithelial Cells / Q000378:metabolism / Q000382:microbiology*; D005434:Flow Cytometry; D006388:Hemagglutinins / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D039081:Integrin alpha5beta1 / Q000378:metabolism; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis* / Q000235:genetics; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism*; D037342:Pertussis Toxin / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation; D010566:Virulence Factors, Bordetella / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D018829:Adhesins, Bacterial; D002457:Cell Extracts; D004268:DNA-Binding Proteins; D006388:Hemagglutinins; D020672:I-kappa B Proteins; D039081:Integrin alpha5beta1; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D010566:Virulence Factors, Bordetella; C065273:filamentous hemagglutinin adhesin, Bordetella pertussis; D018799:Intercellular Adhesion Molecule-1; D000072000:NF-KappaB Inhibitor alpha; D037342:Pertussis Toxin