Antibodies produced against a fragment of filamentous haemagglutinin (FHA) of Bordetella pertussis are able to inhibit hemagglutination induced by the whole adhesin.
Filamentous hemagglutinin adhesin (FHA) is important for the adherence of Bordetella pertussis to the host ciliary epithelial cells of the respiratory tract. Several binding domains have been characterized in the FHA molecule. For example, an putative heparin-binding domain of FHA was previously located in the FHA(442-863) region. In this work, the HEP fragment, corresponding to FHA(430-873) was amplified by PCR and subcloned in an Escherichia coli expression plasmid. Purified recombinant HEP was used to produce polyclonal antibodies in mice that were able to recognize HEP and FHA in ELISA and in Western-blot assays. Although recombinant HEP displayed low ability to bind heparin and no hemagglutination activity, the anti-HEP antibodies were able to inhibit FHA mediated hemagglutination activity in goose erythrocytes. These results indicate that other amino acid residues that are not present in the FHA(430-873) fragment may be necessary for heparin binding. Further studies to address the immunogenic response against HEP are also required.
Colombi
Débora
D
Centro de Biotecnologia, Instituto Butantan, Av. Vital Brasil, 1500 Butantan, CEP 05503-900 São Paulo, SP, Brazil.
Horton
Denise S P Q
DS
Oliveira
Maria Leonor S
ML
Sakauchi
Maria Aparecida
MA
Ho
Paulo L
PL
D018829:Adhesins, Bacterial / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000918:Antibody Specificity; D001886:Bordetella pertussis / Q000276:immunology*; D003001:Cloning, Molecular; D004912:Erythrocytes / Q000276:immunology / Q000382:microbiology; D005260:Female; D006384:Hemagglutination / Q000276:immunology*; D006388:Hemagglutinins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D006493:Heparin / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D010566:Virulence Factors, Bordetella / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D014917:Whooping Cough / Q000517:prevention & control
D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D006388:Hemagglutinins; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; C065273:filamentous hemagglutinin adhesin, Bordetella pertussis; D006493:Heparin