Determination of the components of immune complexes made in vitro with antigens derived from Pseudomonas aeruginosa.
A method for identification of the components of immune complexes would increase our understanding of the pathogenesis of chronic diseases such as Pseudomonas aeruginosa lung infection in cystic fibrosis. Capillary tube, gel diffusion and turbidimetric methods of determining immune complex formation were investigated with antigens from P. aeruginosa and the homologous rabbit antisera. Visible complexes were formed in the first two methods with flagella antigens. Purified lipopolysaccharide from P. aeruginosa would not form visible precipitates and a rapid and economical turbidimetric method was developed with 96-well microtiter plates. Larger quantities of immune complexes were formed in vitro with antigen/antibody ratios determined by the above methods. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting (Western blotting) were evaluated and found to be useful in determining the antigen and antibody components of these immune complexes.
Kronborg
G
G
Department of Clinical Microbiology, Rigshospitalet, Copenhagen, Denmark.
Fomsgaard
A
A
Shand
G H
GH
Høiby
N
N
D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis* / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000032:analysis*; D000942:Antigens, Bacterial / Q000032:analysis* / Q000276:immunology; D015153:Blotting, Western; D010861:Fimbriae, Bacterial / Q000276:immunology; D005407:Flagella / Q000276:immunology; D005779:Immunodiffusion; D008070:Lipopolysaccharides / Q000276:immunology; D008854:Microscopy, Electron; D008970:Molecular Weight; D011550:Pseudomonas aeruginosa / Q000276:immunology*; D011817:Rabbits
D000907:Antibodies, Bacterial; D000936:Antigen-Antibody Complex; D000942:Antigens, Bacterial; D008070:Lipopolysaccharides