Affiliation:
1. Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115
Abstract
ABSTRACT
Numerous studies have reported that asialo-GM
1
, gangliotetraosylceramide, or moieties serve as epithelial cell receptors for
Pseudomonas aeruginosa
. Usually this interaction is confirmed with antibodies to asialo-GM
1
. However, few, if any, of these reports have evaluated the binding of fresh clinical isolates of
P. aeruginosa
to asialo-GM
1
or the specificity of the antibodies for the asialo-GM
1
antigen. We confirmed that asialo-GM
1
dissolved in dimethyl sulfoxide could be added to the apical membrane of Madin-Darby canine kidney cells growing as a polarized epithelium on Transwell membranes (J. C. Comolli, L. L. Waite, K. E. Mostov, and J. N. Engel, Infect. Immun. 67:3207–3214, 1999) and that such treatment enhanced the binding of
P. aeruginosa
strain PA103. However, no other
P. aeruginosa
strain, including eight different clinical isolates, exhibited enhanced binding to asialo-GM
1
-treated cells. Studies with commercially available antibodies to asialo-GM
1
showed that these preparations had high titers of antibody to
P. aeruginosa
antigens, including whole cells, purified lipopolysaccharide (LPS), and pili. Inhibition studies showed that adsorption of an antiserum to asialo-GM
1
with
P. aeruginosa
cells could remove the reactivity of antibodies to asialo-GM
1
, and adsorption of this serum with asialo-GM
1
removed antibody binding to
P. aeruginosa
LPS. Antibodies in sera raised to asialo-GM
1
were observed to bind to
P. aeruginosa
cells by immunoelectron microscopy. Antibodies to asialo-GM
1
inhibited formation of a biofilm by
P. aeruginosa
in the absence of mammalian cells, indicating a direct inhibition of bacterial cell-cell interactions. These findings demonstrate that asialo-GM
1
is not a major cellular receptor for clinical isolates of
P. aeruginosa
and that commercially available antibodies raised to this antigen contain high titers of antibody to multiple
P. aeruginosa
antigens, which do not interfere with the binding of
P. aeruginosa
to mammalian cells but possibly interfere with the binding of
P. aeruginosa
cells to each other.
Publisher
American Society for Microbiology
Subject
Infectious Diseases,Immunology,Microbiology,Parasitology
Cited by
47 articles.
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