HEp-2 Cell Adherence, Actin Aggregation, and Intimin Types of Attaching and Effacing Escherichia coli Strains Isolated from Healthy Infants in Germany and Australia

ABSTRACT Fecal samples from healthy children under 2 years of age living in Berlin, Germany (205 infants), and Melbourne, Australia (184 infants), were investigated for the presence of attaching and effacing (AE) Escherichia coli (AEEC) strains by screening for eae (intimin) genes. Twenty-seven AEEC strains were isolated from 14 children (7.6%) from Melbourne and from 12 children (5.9%) from Berlin. The 27 AEEC strains were classified as enterohemorrhagic E. coli (one strain, producing Shiga toxin 1), typical enteropathogenic E. coli (EPEC) (one strain carrying an EPEC adherence factor [EAF] plasmid), and atypical EPEC (25 strains negative for Shiga toxins and EAF plasmids). The AEEC were divided into 18 different serotypes, O-nontypeable and O-rough strains. Typing of their intimin genes revealed the presence of intimin α in 6 strains, intimin β in 11 strains, intimin γ in 7 strains, intimin ζ in 2 strains, and intimin η in one strain. Analysis of HEp-2 cell adherence showed diffuse adherence or localized adherence-like patterns in 26 AEEC strains; local adherence was found only with the EAF-positive strain. Ten AEEC strains showed an AE property with the fluorescent actin staining (FAS) test. The introduction of an EAF plasmid (pMAR7) converted 11 FAS-negative AEEC strains to FAS positive and increased the FAS reaction in six FAS-positive AEEC strains, indicating that the genes needed for the AE phenotype were functional in these strains. Our finding indicates that atypical EPEC strains could play a double role as strains that naturally immunize against intimin in humans and as reservoirs for new emerging human pathogenic EPEC strains.

[1]  L. Beutin,et al.  Serotypes of Escherichia coli isolated from healthy infants in Berlin, Germany and Melbourne, Australia. , 2003, Comparative immunology, microbiology and infectious diseases.

[2]  L. Beutin,et al.  Genetic Diversity of Intimin Genes of Attaching and Effacing Escherichia coli Strains , 2002, Journal of Clinical Microbiology.

[3]  M. Thomson,et al.  Tissue Tropism of Enteropathogenic Escherichia coli Strains Belonging to the O55 Serogroup , 2002, Infection and Immunity.

[4]  S. H. Fabbricotti,et al.  Comparison of DNA Hybridization and PCR Assays for Detection of Putative Pathogenic Enteroadherent Escherichia coli , 2002, Journal of Clinical Microbiology.

[5]  T. Whittam,et al.  Molecular Evolution of the Intimin Gene in O111 Clones of Pathogenic Escherichia coli , 2002, Journal of bacteriology.

[6]  V. Souza,et al.  The elements of the locus of enterocyte effacement in human and wild mammal isolates of Escherichia coli: evolution by assemblage or disruption? , 2001, Microbiology.

[7]  I. Okeke,et al.  Comparative Sequence Analysis of the Plasmid-Encoded Regulator of Enteropathogenic Escherichia coliStrains , 2001, Infection and Immunity.

[8]  J. Foster,et al.  An activator of glutamate decarboxylase genes regulates the expression of enteropathogenic Escherichia coli virulence genes through control of the plasmid‐encoded regulator, Per , 2001, Molecular microbiology.

[9]  L. Beutin,et al.  Occurrence of Shiga toxin-producing Escherichia coli in fecal samples from children with diarrhea and from healthy zebu cattle in Uganda. , 2001, International journal of food microbiology.

[10]  E. Boedeker,et al.  Complete Nucleotide Sequence and Analysis of the Locus of Enterocyte Effacement from Rabbit Diarrheagenic Escherichia coli RDEC-1 , 2001, Infection and Immunity.

[11]  G. Dougan,et al.  Site‐directed mutagenesis of intimin α modulates intimin‐mediated tissue tropism and host specificity , 2001, Molecular microbiology.

[12]  J. R. Andrade,et al.  Phenotypic and genotypic characteristics of Escherichia coli strains of non-enteropathogenic E. coli (EPEC) serogroups that carry EAE and lack the EPEC adherence factor and Shiga toxin DNA probe sequences. , 2001, The Journal of infectious diseases.

[13]  J. Puente,et al.  Transcriptional regulation of type III secretion genes in enteropathogenic Escherichia coli: Ler antagonizes H‐NS‐dependent repression , 2001, Molecular microbiology.

[14]  L. Trabulsi,et al.  10.321/eid0805.Typical and Atypical Enteropathogenic Escherichia coli , 2002, Emerging infectious diseases.

[15]  F. Grimont,et al.  Identification of Escherichia coli flagellar types by restriction of the amplified fliC gene. , 2000, Research in microbiology.

[16]  L. Beutin,et al.  Investigation of human infections with verocytotoxin-producing strains of Escherichia coli (VTEC) belonging to serogroup O118 with evidence for zoonotic transmission , 2000, Epidemiology and Infection.

[17]  B. Finlay,et al.  Exploitation of host cells by enteropathogenic Escherichia coli. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[18]  J. Nougayrède,et al.  Role of Tir and Intimin in the Virulence of Rabbit Enteropathogenic Escherichia coli Serotype O103:H2 , 2000, Infection and Immunity.

[19]  R. Robins-Browne,et al.  Identification of a novel genetic locus that is required for in vitro adhesion of a clinical isolate of enterohaemorrhagic Escherichia coli to epithelial cells , 2000, Molecular microbiology.

[20]  A. Caprioli,et al.  Typing of Intimin Genes in Human and Animal Enterohemorrhagic and Enteropathogenic Escherichia coli: Characterization of a New Intimin Variant , 2000, Infection and Immunity.

[21]  G. Dougan,et al.  Antibody response of patients infected with verocytotoxin-producing Escherichia coli to protein antigens encoded on the LEE locus. , 2000, Journal of medical microbiology.

[22]  B. China,et al.  Comparison of eae, tir, espA and espB genes of bovine and human attaching and effacing Escherichia coli by multiplex polymerase chain reaction. , 1999, FEMS microbiology letters.

[23]  G. Schoolnik,et al.  A novel chromosomal locus of enteropathogenic Escherichia coli (EPEC), which encodes a bfpT‐regulated chaperone‐like protein, TrcA, involved in microcolony formation by EPEC , 1999, Molecular microbiology.

[24]  V. Sperandio,et al.  The Per regulon of enteropathogenic Escherichia coli : identification of a regulatory cascade and a novel transcriptional activator, the locus of enterocyte effacement (LEE)‐encoded regulator (Ler) , 1999, Molecular microbiology.

[25]  V. Sperandio,et al.  Virulence properties of atypical EPEC strains. , 1999, Journal of medical microbiology.

[26]  G. Dougan,et al.  Enteropathogenic and enterohaemorrhagic Escherichia coli : more subversive elements , 1998, Molecular microbiology.

[27]  P. Manning,et al.  Translocated Intimin Receptors (Tir) of Shiga-Toxigenic Escherichia coli Isolates Belonging to Serogroups O26, O111, and O157 React with Sera from Patients with Hemolytic-Uremic Syndrome and Exhibit Marked Sequence Heterogeneity , 1998, Infection and Immunity.

[28]  Nicole T. Perna,et al.  Molecular Evolution of a Pathogenicity Island from Enterohemorrhagic Escherichia coli O157:H7 , 1998, Infection and Immunity.

[29]  T. McDaniel,et al.  The complete sequence of the locus of enterocyte effacement (LEE) from enteropathogenic Escherichia coli E2348/69 , 1998 .

[30]  G. Dougan,et al.  Detection of Intimins α, β, γ, and δ, Four Intimin Derivatives Expressed by Attaching and Effacing Microbial Pathogens , 1998, Journal of Clinical Microbiology.

[31]  H. Karch,et al.  Diffusely Adhering Escherichia coliStrains Induce Attaching and Effacing Phenotypes and Secrete Homologs of Esp Proteins , 1998, Infection and Immunity.

[32]  James P. Nataro,et al.  Diarrheagenic Escherichia coli , 1998, Clinical Microbiology Reviews.

[33]  G. Dougan,et al.  Detection of intimins alpha, beta, gamma, and delta, four intimin derivatives expressed by attaching and effacing microbial pathogens. , 1998, Journal of clinical microbiology.

[34]  J. Besser,et al.  An outbreak of foodborne illness caused by Escherichia coli O39:NM, an agent not fitting into the existing scheme for classifying diarrheogenic E. coli. , 1997, The Journal of infectious diseases.

[35]  B. Finlay,et al.  Enteropathogenic E. coli (EPEC) Transfers Its Receptor for Intimate Adherence into Mammalian Cells , 1997, Cell.

[36]  M. Yamazaki,et al.  eaeA genes in Escherichia coli derived from Japanese patients with sporadic diarrhea. , 1997, Kansenshogaku zasshi. The Journal of the Japanese Association for Infectious Diseases.

[37]  T. Whittam,et al.  Genetic and phenotypic analysis of Escherichia coli with enteropathogenic characteristics isolated from Seattle children. , 1997, The Journal of infectious diseases.

[38]  C. Le Bouguénec,et al.  Enteroadherent Escherichia coli and diarrhea in children: a prospective case-control study , 1996, Journal of clinical microbiology.

[39]  L. Beutin,et al.  Rapid detection and isolation of shiga-like toxin (verocytotoxin)-producing Escherichia coli by direct testing of individual enterohemolytic colonies from washed sheep blood agar plates in the VTEC-RPLA assay , 1996, Journal of clinical microbiology.

[40]  G. Schoolnik,et al.  Cloning and characterization of bfpTVW, genes required for the transcriptional activation of bfpA in enteropathogenic Escherichia coli , 1996, Molecular microbiology.

[41]  C. Gyles,et al.  Detection of toxin genes in Escherichia coli isolated from normal dogs and dogs with diarrhea. , 1995, Canadian journal of veterinary research = Revue canadienne de recherche veterinaire.

[42]  J. Kaper,et al.  A plasmid-encoded regulatory region activates chromosomal eaeA expression in enteropathogenic Escherichia coli , 1995, Infection and immunity.

[43]  L. Riley,et al.  Identification of enteropathogenic Escherichia coli by PCR-based detection of the bundle-forming pilus gene , 1995, Journal of clinical microbiology.

[44]  L. Beutin,et al.  Virulence markers of Shiga-like toxin-producing Escherichia coli strains originating from healthy domestic animals of different species , 1995, Journal of clinical microbiology.

[45]  B. Rowe,et al.  Catabolite repression of the adhesion of Vero cytotoxin-producing Escherichia coli of serogroups O157 and O111. , 1995, Microbial pathogenesis.

[46]  T. McDaniel,et al.  A genetic locus of enterocyte effacement conserved among diverse enterobacterial pathogens. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[47]  L. Beutin,et al.  Nucleotide sequence analysis of enteropathogenic Escherichia coli (EPEC) adherence factor probe and development of PCR for rapid detection of EPEC harboring virulence plasmids , 1994, Journal of clinical microbiology.

[48]  H. Karch,et al.  Prevalence of attaching and effacing Escherichia coli in stool samples from patients and controls. , 1994, Zentralblatt fur Bakteriologie : international journal of medical microbiology.

[49]  K. Jarvis,et al.  Distribution of the bundle-forming pilus structural gene (bfpA) among enteropathogenic Escherichia coli. , 1993, The Journal of infectious diseases.

[50]  J. Nataro,et al.  A plasmid‐encoded type IV fimbrial gene of enteropathogenic Escherichia coli associated with localized adherence , 1992, Molecular microbiology.

[51]  J. Kaper,et al.  Enteropathogenic Escherichia coli. , 1992 .

[52]  J. Kaper,et al.  Enteropathogenic Escherichia coli , 1992, Infection and immunity.

[53]  G. Schoolnik,et al.  An inducible bundle-forming pilus of enteropathogenic Escherichia coli. , 1991, Science.

[54]  S. Moseley,et al.  HeLa cell adherence, actin aggregation, and invasion by nonenteropathogenic Escherichia coli possessing the eae gene , 1991, Infection and immunity.

[55]  M. McKee,et al.  Two copies of Shiga-like toxin II-related genes common in enterohemorrhagic Escherichia coli strains are responsible for the antigenic heterogeneity of the O157:H- strain E32511 , 1991, Infection and immunity.

[56]  A. Phillips,et al.  Screening for enteropathogenic Escherichia coli in infants with diarrhea by the fluorescent-actin staining test , 1991, Infection and immunity.

[57]  L. Beutin,et al.  Clonal diversity and virulence factors in strains of Escherichia coli of the classic enteropathogenic serogroup O114. , 1990, The Journal of infectious diseases.

[58]  B. Rowe,et al.  Properties of strains of Escherichia coli O26:H11 in relation to their enteropathogenic or enterohemorrhagic classification. , 1990, The Journal of infectious diseases.

[59]  J. Yu,et al.  A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[60]  M. Levine,et al.  A sensitive and specific DNA probe to identify enteroaggregative Escherichia coli, a recently discovered diarrheal pathogen. , 1990, The Journal of infectious diseases.

[61]  S. Moseley,et al.  Molecular characterization of a fimbrial adhesin, F1845, mediating diffuse adherence of diarrhea-associated Escherichia coli to HEp-2 cells , 1989, Journal of bacteriology.

[62]  B. Rowe,et al.  Properties of adherence factor plasmids of enteropathogenic Escherichia coli and the effect of host strain on expression of adherence to HEp-2 cells. , 1989, Journal of general microbiology.

[63]  P. Williams,et al.  Actin accumulation at sites of bacterial adhesion to tissue culture cells: basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli , 1989, Infection and immunity.

[64]  J. Nataro,et al.  Characterization of plasmids encoding the adherence factor of enteropathogenic Escherichia coli , 1987, Infection and immunity.

[65]  K. Bettelheim,et al.  New method of serotyping Escherichia coli: implementation and verification , 1987, Journal of clinical microbiology.

[66]  I. Ørskov,et al.  2 Serotyping of Escherichia coli , 1984 .

[67]  M. Levine,et al.  Plasmid‐Mediated Adhesion in Enteropathogenic Escherichia coli , 1983, Journal of pediatric gastroenterology and nutrition.

[68]  G. Dougan,et al.  Temperature-sensitive mutants of equine arteritis virus , 1980 .

[69]  Larry K. Pickering,et al.  Infections of the Gastrointestinal Tract , 1980, Current Topics in Infectious Disease.