title: Flagellar Cap Protein FliD Mediates Adherence of Atypical Enteropathogenic Escherichia coli to Enterocyte Microvilli.
abstract: The expression of flagella correlates with different aspects of bacterial pathogenicity, ranging from adherence to host cells to activation of inflammatory responses by the innate immune system. In the present study, we investigated the role of flagella in the adherence of an atypical enteropathogenic Escherichia coli (aEPEC) strain (serotype O51:H40) to human enterocytes. Accordingly, isogenic mutants deficient in flagellin (FliC), the flagellar structural subunit; the flagellar cap protein (FliD); or the MotAB proteins, involved in the control of flagellar motion, were generated and tested for binding to differentiated Caco-2 cells. Binding of the aEPEC strain to enterocytes was significantly impaired in strains with the fliCa nd fliD genes deleted, both of which could not form flagella on the bacterial surface. A nonmotile but flagellated MotAB mutant also showed impaired adhesion to Caco-2 cells. In accordance with these observations, adhesion of a EPEC strain 1711-4 to Caco-2 cells was drastically reduced after the treatment of Caco-2 cells with purified FliD. In addition, incubation of a EPEC bacteria with specific anti-FliD serum impaired binding to Caco-2 cells. Finally, incubation of Caco-2 cells with purified FliD, followed by immunolabeling, showed that the protein was specifically bound to the microvillus tips of differentiated Caco-2 cells. The a EPEC FliD or anti-FliD serum also reduced the adherence of prototype typical enteropathogenic, enterohemorrhagic, and enterotoxigenic E. coli strains to Caco-2 cells. In conclusion, our findings further strengthened the role of flagella in the adherence of a EPEC to human enterocytes and disclosed the relevant structural and functional involvement of FliD in the adhesion process.
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brasil.
authors.forename: Suely C F
authors.initials: SCF
authors.lastname: Sampaio
authors.affiliation: Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brasil.
authors.forename: Wilson B
authors.initials: WB
authors.lastname: Luiz
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brasil.
authors.forename: Mônica A M
authors.initials: MAM
authors.lastname: Vieira
authors.affiliation: Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brasil.
authors.forename: Rita C C
authors.initials: RCC
authors.lastname: Ferreira
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brasil.
authors.forename: Bruna G
authors.initials: BG
authors.lastname: Garcia
authors.affiliation: Centro de Microscopia Eletrônica, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brasil.
authors.forename: Rita
authors.initials: R
authors.lastname: Sinigaglia-Coimbra
authors.affiliation: Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, São Paulo, Brasil.|Instituto Fleury de Ensino e Pesquisa, São Paulo, Brasil.
authors.forename: Jorge L M
authors.initials: JLM
authors.lastname: Sampaio
authors.affiliation: Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brasil.
authors.forename: Luís C S
authors.initials: LCS
authors.lastname: Ferreira
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brasil tatg.amaral@unifesp.br.
authors.forename: Tânia A T
authors.identifier: http://orcid.org/0000-0002-4525-8705
authors.initials: TAT
authors.lastname: Gomes
mesh_terms: D000818:Animals; D000906:Antibodies; D001422:Bacterial Adhesion / Q000502:physiology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018938:Caco-2 Cells; D020895:Enterocytes / Q000382:microbiology* / Q000502:physiology; D054308:Enteropathogenic Escherichia coli / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D008871:Microvilli / Q000502:physiology*; D009154:Mutation; D011817:Rabbits; D011994:Recombinant Proteins
chemical_list: D000906:Antibodies; D001426:Bacterial Proteins; D011994:Recombinant Proteins; C076733:FlaD protein, Bacteria
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