Analysis and modelling of Pseudomonas aeruginosa adherence to human buccal epithelial cells

1997 ◽  
Vol 8 (4-5) ◽  
pp. 267-277
Author(s):  
Anne M.C. Cerf ◽  
Jean-Paul Dehaye ◽  
Michel J. Devleeschouwer
1980 ◽  
Vol 29 (3) ◽  
pp. 1146-1151 ◽  
Author(s):  
D E Woods ◽  
D C Straus ◽  
W G Johanson ◽  
V K Berry ◽  
J A Bass

Adherence of Pseudomonas aeruginosa organisms to the upper respiratory epithelium of seriously ill patients in vitro is correlated with subsequent colonization of the respiratory tract by this opportunistic pathogen. The role of pili in the attachment to epithelial cells of P. aeruginosa was studied in an in vitro system employing human buccal epithelial cells and P. aeruginosa pretreated by various means. Pretreatment of the bacteria with proteases, heat, or Formalin caused a significant decrease in adherence. A decrease when compared with controls was also noted in the adherence of P. aeruginosa organisms to buccal epithelial cells preincubated with purified pili prepared from the strain used for adherence testing; however, pili prepared from a heterologous strain failed to block adherence. Similar results were obtained in serological studies when antisera to purified pili prepared from the strain used for adherence testing decreased adherence, whereas heterologous antiserum to pili did not decrease adherence. From these results it appears that pili mediate the adherence of P. aeruginosa organisms to human buccal epithelial cells.


1983 ◽  
Vol 5 (Supplement_5) ◽  
pp. S846-S851 ◽  
Author(s):  
Donald E. Woods ◽  
David C. Straus ◽  
W. G. Johanson ◽  
Joe A. Bass

1986 ◽  
Vol 32 (2) ◽  
pp. 160-166 ◽  
Author(s):  
P. Doig ◽  
A. L. Franklin ◽  
R. T. Irvin

The binding of outer membrane (OM) ghosts derived from Pseudomonas aeruginosa strain 492c to human buccal epithelial cells (BECs) was examined. Electron microscopic examination of the binding of OM ghosts to BECs revealed direct OM ghost–BEC interaction. Equilibrium analysis of the binding of OM ghosts to trypsinized BECs employing the Langmuir adsorption isotherm indicated the number of binding sites (iV) to be 1.3 × 10−1 μg protein per BEC with an apparent association constant (Ka) of 3.4 × 10−2 mL/μg protein. The Langmuir analysis of binding of OM ghosts to untrypsinized BECs was complex, suggesting two possible classes of receptors, a high affinity–low copy number class (Ka, 1.8 × 10−2 mL/μg protein; N, 8.6 × 10−5 μg protein per BEC) and a low affinity – high copy number class(Afa, 3.7 × 10−3 mL/μg protein; N, 9.2 × 10−4 μg protein per BEC). Sugar inhibition studies incorporating D-galactose enhanced binding to each BEC type. N-Acetylneuraminic acid and N-acetyl-glucosamine both enhanced binding of OM ghosts to untrypsinized BECs, while inhibiting binding to trypsinized BECs. D-Arabinose inhibited binding to both BEC types. Binding of OM ghosts to both BEC types was greatly inhibited by D-fucose, while L-fucose only greatly inhibited binding to untrypsinized BECs. These sugar inhibition data demonstrated a difference in the binding of OM ghosts to trypsinized and untrypsinized BECs and possibly reveal the nature of the receptor(s), free of possible bacterial metabolic effects. These data indicated that OM ghosts from 492c appear to bind to BECs in a similar manner to the intact bacteria and represent a simple model system to study the adhesion of P. aeruginosa to BECs.


2012 ◽  
Vol 49 (1) ◽  
pp. 59-64
Author(s):  
Katarzyna Wolska ◽  
Barbara Kot ◽  
Halina Mioduszewska ◽  
Cezary Sempruch ◽  
Lidia Borkowska ◽  
...  

Abstract This study shows an association between the frequency of the nan1 gene (encoding neuraminidase) among 62 clinical Pseudomonas aeruginosa isolates and adhesion of these bacteria to human buccal epithelial cells. The 52 strains in which the gene was present (83.9%) were characterized by a higher adhesiveness (the mean number of adhering bacteria was 23.51 per cell) than strains in which the gene was not detected (16.23 per cell) and the difference was significant (P = 0.009, Mann-Whitney U test). Thus we found that the nan1 gene may play a role in the binding of clinical P. aeruginosa strains to buccal cells.


1989 ◽  
Vol 35 (12) ◽  
pp. 1141-1145 ◽  
Author(s):  
Peter Doig ◽  
William Paranchych ◽  
Parimi A. Sastry ◽  
Randall T. Irvin

Adherence of Pseudomonas aeruginosa to a patient's epithelial surface is thought to be an important first step in the infection process. Pseudomonas aeruginosa is capable of attaching to epithelial cells via its pili, yet little is known about the epithelial receptors of this adhesin. Using nitrocellulose replicas of polyacrylamide gels of solubilized human buccal epithelial cells (BECs), glycoproteins (Mz: 82 000, and four bands between 40 000 and 50 000) that bound purified pili from P. aeruginosa strain K (PAK) were identified by immunoblotting with a pilus-specific monoclonal antibody that does not affect pilus binding to BECs (PK3B). All pilus-binding glycoproteins were surface localized, as determined by surface radioiodination of intact BECs. Binding of pili to all of the glycoproteins was inhibited by Fab fragments of monoclonal antibody PK99H, which inhibits PAK pili binding to BECs by binding to or near the binding domain of the pilus, but not by Fab fragments of monoclonal antibody PK41C, which binds to PAK pilin but does not inhibit pili binding to BECs, demonstrating that pilus binding to these glycoproteins is likely via the same region of the pilus that binds to intact BECs. Periodate oxidation of the blot eliminated pili binding to all glycoproteins, indicating that a carbohydrate moiety is an important determinant for pilus-binding activity. However, not all of the glycoproteins exhibited the same degree of sensitivity to periodate oxidation. Furthermore, monosaccharide inhibition of pilus binding to BECs implicated L-fucose and N-acetylneuraminic acid as receptor moieties.Key words: Pseudomonas aeruginosa, pili, receptor, adhesion.


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