title: Staphylococcus aureus clumping factor B (ClfB) promotes adherence to human type I cytokeratin 10: implications for nasal colonization.
abstract: Staphylococcus aureus is an important cause of sepsis in both community and hospital settings, a major risk factor for which is nasal carriage of the bacterium. Eradication of carriage by topical antibiotics reduces sepsis rates in high-risk individuals, an important strategy for the reduction of nosocomial infection in targeted patient populations. Understanding the mechanisms by which S. aureus adheres to nasal epithelial cells in vivo may lead to alternative methods of decolonization that do not rely on sustained antimicrobial susceptibility. Here, we demonstrate for the first time that the S. aureus surface-expressed protein, clumping factor B (ClfB), promotes adherence to immobilized epidermal cytokeratins in vitro. By expressing a range of S. aureus adhesins on the surface of the heterologous host Lactococcus lactis, we demonstrated that adherence to epidermal cytokeratins was conferred by ClfB. Adherence of wild-type S. aureus was inhibited by recombinant ClfB protein or anti-ClfB antibodies, and S. aureus mutants defective in ClfB adhered poorly to epidermal cytokeratins. Expression of ClfB promoted adherence of L. lactis to human desquamated nasal epithelial cells, and a mutant of S. aureus defective in ClfB had reduced adherence compared with wild type. ClfB also promoted adherence of L. lactis cells to a human keratinocyte cell line. Cytokeratin 10 molecules were shown by flow cytometry to be exposed on the surface of both desquamated nasal epithelial cells and keratinocytes. Cytokeratin 10 was also detected on the surface of desquamated human nasal cells using immunofluorescence, and recombinant ClfB protein was shown to bind to cytokeratin K10 extracted from these cells. We also showed that ClfB is transcribed by S. aureus in the human nares. We propose that ClfB is a major determinant in S. aureus nasal colonization.
authors.affiliation: Moyne Institute of Preventive Medicine, Department of Microbiology, Trinity College Dublin, Dublin 2, Ireland.
authors.forename: Louise M
authors.initials: LM
authors.lastname: O'Brien
authors.forename: Evelyn J
authors.initials: EJ
authors.lastname: Walsh
authors.forename: Ruth C
authors.initials: RC
authors.lastname: Massey
authors.forename: Sharon J
authors.initials: SJ
authors.lastname: Peacock
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Foster
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism*; D001422:Bacterial Adhesion / Q000502:physiology*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D053550:Keratin-10; D015603:Keratinocytes / Q000378:metabolism; D007633:Keratins / Q000378:metabolism*; D013294:Lactococcus lactis / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D009296:Nasal Cavity / Q000166:cytology / Q000382:microbiology*; D011485:Protein Binding; D020033:Protein Isoforms / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013211:Staphylococcus aureus / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D018829:Adhesins, Bacterial; C507493:KRT10 protein, human; D020033:Protein Isoforms; D011994:Recombinant Proteins; C434058:clfB protein, Staphylococcus aureus; D053550:Keratin-10; D007633:Keratins