Monoclonal antibody S60-4-14 reveals diagnostic potential in the identification of Pseudomonas aeruginosa in lung tissues of cystic fibrosis patients.
The lipopolysaccharide (LPS) of Pseudomonas aeruginosa has been identified to contain an inner-core structure expressing a Pseudomonas-specific epitope. This target structure is characterized by a highly phosphorylated and 7-O-carbamoyl-l-glycero-alpha-d-manno-heptopyranose (CmHep) and was found to be present in all human-pathogenic Pseudomonas species of the Palleroni (RNA)-classification I scheme. We raised and selected the monoclonal antibody S60-4-14 (mAb S60-4-14, subtype IgG1) from mice immunized with heat-killed Pseudomonas bacteria. The epitope of this mAb was found to reside in the inner-core structure of P. aeruginosa and, hence, successfully evaluated for the immunohistochemical detection of P. aeruginosa in formalin- or HOPE-fixed (Hepes-glutamic acid buffer-mediated organic solvent protection effect) and paraffin-embedded human lung tissue slices. Lung specimens, mainly from explanted lungs of cystic fibrosis (CF) patients, as well as P. aeruginosa isolates from patients suffering from CF and patients with extrapulmonar Pseudomonas infections were investigated by PCR, immunohistochemistry, and Western blot analysis with mAb S60-4-14. The results revealed an unequivocal coincidence of PCR and immunohistochemistry. Together with the Western blot results mAb S60-4-14 displays a potential diagnostic tool for the specific identification of P. aeruginosa in infected lungs of CF.
Schmengler
Kai
K
Division of Immunochemistry, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Parkallee 4a, 23845 Borstel, Germany.
Goldmann
Torsten
T
Brade
Lore
L
Sánchez Carballo
Patricia M
PM
Albrecht
Sandra
S
Brade
Helmut
H
Kosma
Paul
P
Sahly
Hany
H
Hauber
Hans-Peter
HP
Länger
Florian
F
Meyer
Rudolf
R
Welte
Tobias
T
Zähringer
Ulrich
U
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D002236:Carbohydrate Conformation; D003550:Cystic Fibrosis / Q000150:complications* / Q000382:microbiology* / Q000473:pathology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007150:Immunohistochemistry; D008070:Lipopolysaccharides / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D008168:Lung / Q000382:microbiology*; D010766:Phosphorylation; D011552:Pseudomonas Infections / Q000150:complications* / Q000175:diagnosis*; D011550:Pseudomonas aeruginosa / Q000276:immunology* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D008070:Lipopolysaccharides