Biological activity of a human monoclonal antibody to Bordetella pertussis lipooligosaccharide.
The heterohybridoma cell line HBp2 secreting human monoclonal antibody (hMAb) directed against Bordetella pertussis was generated by fusing SP2/HPT heteromyeloma cells with human spleen lymphocytes, after in vitro stimulation for 6 days. The hybridoma was maintained in culture for more than 1 year with continuous antibody secretion. The hMAb HBp2, an IgM, reacted with untreated and proteinase K-treated B. pertussis outer membrane antigens, whereas the reactivity was lost when the antigen was treated with sodium periodate. Human MAb HBp2 was shown to be specific to B. pertussis LOS by immunoblotting of whole cell extracts after SDS-PAGE. In a dot enzyme immunoassay, HBp2 reacted with all B. pertussis strains and clinical isolates tested except for four atypical variant strains of the LOS B phenotype. Human MAb HBp2 also reacted with a clinical isolate of B. bronchiseptica. No reaction was observed against B. parapertussis and other gram-negative species. Together these studies suggested that HBp2 is reactive with carbohydrate epitopes present on the LOS A. Binding assays with live bacteria demonstrated that hMAb HBp2 reacted with cell surface exposed epitopes on B. pertussis but the antibody did not bind significantly to the surface on intact B. bronchiseptica cells. When examined for bactericidal activity in the presence of complement, hMAb HBp2 showed high lytic capability against B. pertussis while no killing was obtained against B. bronchiseptica. These experiments established that LOS A is a target for human bactericidal antibodies. This antigen merits further investigation as a potentially important component in human immunity to B. pertussis infection.
Brodeur
B R
BR
National Laboratory for Immunology, Laboratory Centre for Disease Control, Ottawa, Ontario, Canada.
Hamel
J
J
Martin
D
D
Rondeau
P
P
D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000920:Antibody-Dependent Cell Cytotoxicity; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology; D001886:Bordetella pertussis / Q000276:immunology*; D000939:Epitopes; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D008070:Lipopolysaccharides; C023023:lipid-linked oligosaccharides