Auli Pere

Occurrence of type-1C fimbriae on Escherichia coli strains isolated from human extraintestinal infections.

Two monoclonal antibodies specific for type-1C fimbriae of Escherichia coli were produced. In enzyme-linked immunosorbent assay and immunoblotting the antibodies, which were of the IgG1 isotype, reacted with type-1C, but not with P or type-1 fimbriae of E. coli strain KS71. Immunoblotting and immunoprecipitation of crude fimbrial extracts from 25 strains invariably gave an apparent molecular weight of 17 000 for the type-1C fimbrillin. A total of 313 E. coli strains, isolated from patients with extraintestinal infection or from faeces of healthy children, were screened for the presence of type-1C fimbriae using both the monoclonal and polyclonal antibodies. Of these, 45 (14%) strains had type-1C fimbriae, with the highest frequency (27%) on strains isolated from patients with pyelonephritis. No faecal strain had type-1C fimbriae, and the frequency on the other diagnostic groups ranged from 11 to 15%. Thus, no direct correlation between type-1C fimbriae and bacterial virulence in human extraintestinal infections was found. Type-1C fimbriae were detected on only a few E. coli serotypes, notably on all O6:K2:H1 and O22:K13:H1 strains tested.

P-fimbriae vaccines. I: Cross reactive antibodies to heterologous P-fimbriae

To test for cross-protective capacity of two different P-fimbriae vaccines we vaccinated baboons with fimbriae purified from eitherEscherichia coli strain ER2 or strain JR1. The vaccinated animals showed elevated antibody titers to P-fimbriae from each of theE. coli strains used, suggesting cross-reactivity as was expected from the results of immunoprecipitation of the fimbriae. Enzyme-linked immunosorbent assay inhibition by heterologous P-fimbriae proved this to be true immunologic cross-reactivity.

Complementation and regulatory interaction between two cloned fimbrial gene clusters of Escherichia coli strain KS71

SummaryComplementation experiments with cloned DNA fragments encoding either the KS71A, the KS71B or the KS71C fimbriae of the pyelonephritogenic Escherichia coli strain KS71 were used to localise the P-fimbrillin genes and to demonstrate regulatory interactions between the cloned genes. The structural genes of the KS71A and KS71B fimbriae were located within a common 1.1 kilobase pair ClaI-SmaI fragment, and it was shown that the gene clusters for these fimbriae could complement each other in trans. The gene cluster encoding the KS71C fimbriae did not complement for the other KS71 fimbriae. A DNA fragment, located near the KS71A fimbrillin gene, was found to enhance the production of the KS71B fimbriae in trans.

Organization of fimbriate cells in colonies of Escherichia coli strain 3040

Immunofluorescence staining with fimbria-specific antibodies was used to study the organization of fimbriate cells in colonies of Escherichia coli strain 3040. The strain has both type-1 and S fimbriae and shows fast phase variation between the fimbrial types. Colonies stained in sectors whose length and number per colony were dependent on the fimbrial phase of progeny cells. It is proposed that such sectors result from fimbrial phase variation.

Fimbriation and P-antigen recognition of Escherichia coli strains harbouring mutated recombinant plasmids encoding fimbrial adhesins of the uropathogenic E. coli strain KS71.

Deletion mutants of recombinant plasmids encoding the KS71B fimbrial antigens of the uropathogenic Escherichia coli strain KS71 (O4:K12) were constructed. The effects of these mutations were tested by transforming the mutated plasmids into non-fimbriated E. coli HB101 cells and testing the transformants for fimbriation and haemagglutination. A deletion transcriptionally upstream from the fimbrial subunit gene increased the expression of KS71B fimbriae. Deletion of the fimbrial subunit gene resulted in non-fimbriated but haemagglutinating transformants, whereas a deletion 6 kb transcriptionally downstream from the subunit gene resulted in non-haemagglutinating but fimbriate transformants, indicating that fimbriation and haemagglutination were genetically separable. We also present evidence suggesting that the fimbrillin and haemagglutinin are physically associated in the wild-type KS71 strain.