Inhibition of adherence of Actinobacillus pleuropneumoniae to porcine respiratory tract cells by monoclonal antibodies directed against LPS and partial characterization of the LPS receptors.
Actinobacillus pleuropneumoniae is the causative agent of porcine fibrinohemorrhagic necrotizing pleuropneumonia. We have previously identified the lipopolysaccharides (LPS) as the major adhesin of A. pleuropneumoniae involved in adherence to porcine respiratory tract cells. In the present study, adherence of A. pleuropneumoniae to porcine tracheal frozen sections was inhibited by homologous monovalent Fab fragments produced from monoclonal antibodies 5.1 G8F10 and 102-G02 directed, respectively, against the A. pleuropneumoniae serotype 1 or serotype 2 O-antigens. These results confirm the important role played by LPS in adherence of A. pleuropneumoniae and suggest that these adhesins might represent good vaccine candidates. We also investigated the presence of A. pleuropneumoniae receptors in tracheal cell preparations from piglets of four different breeds. Using Far-Western binding assays, we identified proteins recognized by whole cells of A. pleuropneumoniae reference strains for serotype 1 and 2, and local isolates belonging to the same serotypes, and also recognized by extracted LPS from both reference strains. We confirmed the proteinaceous nature of these LPS-binding molecules by their staining with Coomassie brilliant blue, sensitivity to proteinase K digestion, resistance to sodium m-periodate oxidation, and their inability to stain with glycoprotein-specific reagents. Four low-molecular-mass bands (14-17 kDa) seemed to correspond to histones. We also identified proteins at Mr 38,500 that could represent putative receptors for A. pleuropneumoniae LPS in swine respiratory tract cells.
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Groupe de Recherche sur les Maladies Infectieuses du Porc, and Département de pathologie et microbiologie, Faculté de médecine vétérinaire, Université de Montréal, C.P. 5000, St-Hyacinthe (Québec), Canada J2S 7C6.
Dubreuil
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JD
Gottschalk
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Archambault
M
M
Jacques
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D016977:Actinobacillus pleuropneumoniae / Q000145:classification / Q000276:immunology / Q000472:pathogenicity*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001422:Bacterial Adhesion; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D018950:Lipopolysaccharide Receptors / Q000737:chemistry; D008070:Lipopolysaccharides / Q000276:immunology*; D016676:Macrophages, Alveolar / Q000166:cytology / Q000382:microbiology; D051379:Mice; D011817:Rabbits; D013552:Swine; D014132:Trachea / Q000166:cytology / Q000382:microbiology*
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides