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Our ultimate goal is to develop strain RB51 as a preferential vector for the delivery of protective antigens of other intracellular pathogens to which the induction of a strong Th1 type of immune response is needed for effective protection. As a first step in that direction, we studied the expression of a foreign reporter protein, \u03b2-galactosidase of<jats:italic>Escherichia coli<\/jats:italic>, and the 65-kDa heat shock protein (HSP65) of<jats:italic>Mycobacterium bovis<\/jats:italic>in strain RB51. We cloned the promoter sequences of<jats:italic>Brucella sodC<\/jats:italic>and<jats:italic>groE<\/jats:italic>genes in pBBR1MCS to generate plasmids pBBSODpro and pBBgroE, respectively. The genes for \u03b2-galactosidase (<jats:italic>lacZ<\/jats:italic>) and HSP65 were cloned in these plasmids and used to transform strain RB51. An enzyme assay in the recombinant RB51 strains indicated that the level of \u03b2-galactosidase expression is higher under the<jats:italic>groE<\/jats:italic>promoter than under the<jats:italic>sodC<\/jats:italic>promoter. In strain RB51 containing pBBgroE\/lacZ, but not pBBSODpro\/lacZ, increased levels of \u03b2-galactosidase expression were observed after subjecting the bacteria to heat shock or following internalization into macrophage-like J774A.1 cells. Mice vaccinated with either of the \u03b2-galactosidase-expressing recombinant RB51 strains developed specific antibodies of predominantly the immunoglobulin G2a (IgG2a) isotype, and in vitro stimulation of their splenocytes with \u03b2-galactosidase induced the secretion of gamma interferon (IFN-\u03b3), but not interleukin-4 (IL-4). A Th1 type of immune response to HSP65, as indicated by the presence of specific serum IgG2a, but not IgG1, antibodies, and IFN-\u03b3, but not IL-4, secretion by the specific-antigen-stimulated splenocytes, was also detected in mice vaccinated with strain RB51 containing pBBgroE\/hsp65. Studies with mice indicated that expression of \u03b2-galactosidase or HSP65 did not alter either the attenuation characteristics of strain RB51 or its vaccine efficacy against<jats:italic>B. abortus<\/jats:italic>2308 challenge.<\/jats:p>","DOI":"10.1128\/iai.68.6.3290-3296.2000","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T10:01:19Z","timestamp":1027764079000},"page":"3290-3296","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":56,"title":["<i>Brucella abortus<\/i>Strain RB51 as a Vector for Heterologous Protein Expression and Induction of Specific Th1 Type Immune Responses"],"prefix":"10.1128","volume":"68","author":[{"given":"Ramesh","family":"Vemulapalli","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061-0342"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongqun","family":"He","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061-0342"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen M.","family":"Boyle","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061-0342"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nammalwar","family":"Sriranganathan","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061-0342"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gerhardt G.","family":"Schurig","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061-0342"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","unstructured":"Acha P. 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