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dc.contributor.authorMa, Jiannengen_US
dc.date.accessioned2014-03-14T21:21:25Z
dc.date.available2014-03-14T21:21:25Z
dc.date.issued1991-12-15en_US
dc.identifier.otheretd-10142005-135819en_US
dc.identifier.urihttp://hdl.handle.net/10919/39935
dc.description.abstractActinobacillus pleuropneumoniae is the etiological agent of contagious swine pleuropneumonia, an economically important disease of the swine industry worldwide. Improved control of this disease requires enhanced understanding of the factors contributing to pathogenesis. The objectives of this study were to investigate the immune response and virulence properties of the 110-kilodalton (110-KDa) hemolysins [hemolysin I (HlyI) and hemolysin II (HlyII)] of A. pleuropneumoniae. Several monoclonal antibodies (MAb) to the hemolysins were developed. An IgGl. MAb (8C2) specific for HlyII, as determined by immunoblotting, was cross-linked to Protein A-Sepharose, and HlyII was purified from serotypes 1 and 5 by immunoaffinity chromatography. An indirect enzyme-linked immunosorbent assay (ELISA) using MAb 8C2, or affinitypurified rabbit IgG to both hemolysins, was developed for detection of swine antibody to one or both hemolysins, respectively. In comparison with the complement fixation test, the ELISA was highly sensitive and specific, and was able to identify animals infected with or exposed to most, if not all, serotypes of A. pleuropneumoniae. Several nonhemolytic mutants of A. pleuropneumoniae serotype 5 were isolated following electroporation of the parent with an hemolysin gene whose open-reading-frame was disrupted with a kanamycin resistance gene. One mutant was characterized for phenotypic and pathogenic properties. Biochemical profiles, growth rate, capsule content, and lipopolysaccharide and whole cell protein electrophoretic profiles of the parent and one of the mutants were similar. The nonhemolytic mutant lacked both HlyI and HlyII proteins in culture supernatant and in whole cell lysates as determined by immunoblot analysis; extracellular and intracellular hemolytic and cytotoxic activity was also absent. The mutant was avirulent in mice and pigs at doses greater than 10 times the lethal dose of the parent. Unlike the parent, the nonhemolytic mutant failed to confer protection against lethal challenge in mice following immunization. Thus, one or both hemolysins are essential for virulence and immunoprotection in A. pleuropneumoniae serotype 5.en_US
dc.format.mediumBTDen_US
dc.publisherVirginia Techen_US
dc.relation.haspartLD5655.V856_1991.M3.pdfen_US
dc.subjectPleuropneumonia Pathogenesis.en_US
dc.subjectSwine Diseases.en_US
dc.subjectActinobacillus pleuropneumoniae.en_US
dc.subjectHemolysis and hemolysins.en_US
dc.subject.lccLD5655.V856 1991.M3en_US
dc.titlePurification, serology and pathogenic role of the 110 kilodalton rtx hemolysins of Actinobacillus pleuropneumoniaeen_US
dc.typeDissertationen_US
dc.contributor.departmentVeterinary Medical Sciencesen_US
dc.description.degreePh. D.en_US
thesis.degree.namePh. D.en_US
thesis.degree.leveldoctoralen_US
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen_US
thesis.degree.disciplineVeterinary Medical Sciencesen_US
dc.contributor.committeechairInzana, Thomas J.en_US
dc.contributor.committeememberBoyle, Stephen M.en_US
dc.contributor.committeememberSchurig, Gerhardt G.en_US
dc.contributor.committeememberSriranganathan, Nammalwaren_US
dc.contributor.committeememberDean, Dennis R.en_US
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-10142005-135819/en_US
dc.date.sdate2005-10-14en_US
dc.date.rdate2005-10-14
dc.date.adate2005-10-14en_US


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