Adherence of Haemophilus influenzae to Human Epithelial Cells

Gilsdorf, Janet R.; Ferrieri, Patricia

doi:10.3109/00365548409070400

Cited by 26 publications

(14 citation statements)

References 18 publications

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“…Enzyme-linked immunoassays were used to quantitate Hib and NT H. influenzae binding to human buccal epithelial cells and human erythrocyte ghosts. Buccal epithelial cells from five donors were collected, prepared as previously described (12), and pooled. Erythrocyte ghosts were prepared (7) from erythrocytes of an Anton-positive adult donor, from cord erythrocytes, and from erythrocytes of two individuals with Lu(a-b-) dominant phenotype.…”

Section: Methodsmentioning

confidence: 99%

Comparison of hemagglutinating pili of Haemophilus influenzae type b with similar structures of nontypeable H. influenzae

et al. 1992

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Thirty-eight clinical isolates of nontypeable Haemophilus influenzae were tested for the presence of hemagglutinating pili similar to those of H. influenzae type b (Hib) that mediate buccal epithelial cell adherence. Four endogenously hemagglutinating (HA') strains were identified, and eight additional HA' variants were obtained from HAstrains by erythrocyte enrichment. All 12 HA' nontypeable H. influenzae isolates bound antisera directed against denatured pilins of Hib, but none bound antisera against assembled native pili of Hib. In erythrocyteand buccal-cell-binding assays, HA' nontypeable H. influenzae binding was reduced compared with HA' Hib binding and was not significantly different from HAnontypeable H. influenzae binding. Both HAand HA' nontypeable H. influenzae binding was increased over binding of HA-Hib. HA' nontypeable H. influenzae strains agglutinated adult erythrocytes that possess the Anton antigen, which is thought to be the receptor for Hib pili, and did not agglutinate cord or Lu(a-b-) dominant erythrocytes, which lack the Anton antigen. Electron microscopy of HAand HA' variants of three nontypeable H. influenzae strains showed few or no surface appendages on the HA-organisms, but piluslike structures were seen on many organisms from two HA' nontypeable H. influenzae strains and on a few organisms from one strain. Thus, nontypeable H. influenzae appears to possess structures that are immunologically similar to the pilins that make up the hemagglutinating pili of Hib. However, nontypeable H. influenzae appears to also possess mechanisms for erythrocyte and buccal cell adherence that are not directly correlated with the presence of a hemagglutinating pilus. 17).

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Section: Methodsmentioning

confidence: 99%

Comparison of hemagglutinating pili of Haemophilus influenzae type b with similar structures of nontypeable H. influenzae

et al. 1992

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“…), and the clinical microbiology laboratories of the University of Minnesota Hospitals (Minneapolis, Minn.) and the C. S. Mott Children's Hospital (Ann Arbor, Mich.). Isolates were stored as previously described (11). Hemagglutination and adherence assays.…”

Section: Methodsmentioning

confidence: 99%

“…The ability of bacteria to mediate agglutination of human type 0 erythrocytes in vitro was determined by using a semi-quantitative microdilution well technique (31). Bacterial adherence to human buccal epithelial cells (BECs) in vitro was assayed as previously described (11), and the results were expressed as the mean (± standard error of the mean) number of adherent bacteria per cell.…”

Section: Methodsmentioning

confidence: 99%

“…Inhibition of adherence assay. The ability of the rabbit anti-M43 pilus Fab fragments to inhibit the in vitro adherence of the Hib test strains to human BECs was examined by using a modification of the previously described adherence assay (11). One hundred microliters of bacteria suspended in PBS (108 CFU/ml) was incubated with 10 ,ul of either nonimmune or anti-pilus Fab fragments (100 ,g of protein per ml of PBS) for 15 min at room temperature on a rotating platform before the addition of 100 ,l of BECs (105 cells per ml of PBS).…”

Section: Methodsmentioning

confidence: 99%

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Structural and serological relatedness of Haemophilus influenzae type b pili

LiPuma

Gilsdorf

1988

Infect Immun

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The structural and serological relatedness of the pilus proteins of several isolates of Haemophilus influenzae type b cultured from patients with invasive disease and from different anatomic sites within the same patient was examined. Epithelial cell-adherent variants of 25 nonadherent parent isolates were obtained by selection for organisms that adhered to human erythrocytes. Outer membrane protein analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of an additional 24to 24.5-kilodalton protein among all adherent variants but absent from all nonadherent parent isolates. Polyclonal rabbit antiserum * Corresponding author. t Present address:

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“…Type b organisms isolated from the blood or cerebrospinal fluid of patients are hemagglutination deficient and do not express pili (13,35). Strains expressing a hemagglutination-associated pilus as well as strains not expressing this pilus have been isolated from the human nasopharynx (10,24,35). Variants of type b strains expressing pili can also be isolated by enrichment for cells which adhere to human erythrocytes (13,35).…”

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confidence: 99%

Molecular cloning, expression, and sequence of the pilin gene from nontypeable Haemophilus influenzae M37

1991

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Nontypeable Haemophilus influenzae M37 adheres to human buccal epithelial cells and exhibits mannose-resistant hemagglutination of human erythrocytes. An isogenic variant of this strain which was deficient in hemagglutination was isolated. A protein with an apparent molecular weight of 22,000 was present in the sodium dodecyl sulfate-polyacrylamide gel profile of sarcosyl-insoluble proteins from the hemagglutination-proficient strain but was absent from the profile of the isogenic hemagglutination-deficient variant. A monoclonal antibody which reacts with the hemagglutination-proficient isolate but not with the hemagglutination-deficient isolate has been characterized. This monoclonal antibody was employed in an affinity column for purification of the protein as well as to screen a genomic library for recombinant clones expressing the gene. Several clones which contained overlapping genomic fragments were identified by reaction with the monoclonal antibody. The gene for the 22-kDa protein was subcloned and sequenced. The gene for the type b pilin from H. influenzae type b strain MinnA was also cloned and sequenced. The DNA sequence of the strain MinnA gene was identical to that reported previously for two other type b strains. The DNA sequence of the strain M37 gene is 77% identical to that of the type b pilin gene, and the derived amino acid sequence is 68% identical to that of the type b pilin.

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Adherence of Haemophilus influenzae to Human Epithelial Cells

Cited by 26 publications

References 18 publications

Comparison of hemagglutinating pili of Haemophilus influenzae type b with similar structures of nontypeable H. influenzae

Comparison of hemagglutinating pili of Haemophilus influenzae type b with similar structures of nontypeable H. influenzae

Structural and serological relatedness of Haemophilus influenzae type b pili

Molecular cloning, expression, and sequence of the pilin gene from nontypeable Haemophilus influenzae M37

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