Immunogenicity of Brucella-extracted and recombinant protein vaccines in CD-I and BALB/c mice

George W. Pugh Jr. From the National Animal Disease Center, Agricultural Research Service, USDA (Pugh, Tabatabai, Bricker, Phillips, Zehr, Belzer), PO Box 70, Ames, IA 50010, and Department of Zoology, Iowa State University (Mayfield), Ames, IA 50011.

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Louisa B. Tabatabai From the National Animal Disease Center, Agricultural Research Service, USDA (Pugh, Tabatabai, Bricker, Phillips, Zehr, Belzer), PO Box 70, Ames, IA 50010, and Department of Zoology, Iowa State University (Mayfield), Ames, IA 50011.

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Betsy J. Bricker From the National Animal Disease Center, Agricultural Research Service, USDA (Pugh, Tabatabai, Bricker, Phillips, Zehr, Belzer), PO Box 70, Ames, IA 50010, and Department of Zoology, Iowa State University (Mayfield), Ames, IA 50011.

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John E. Mayfield From the National Animal Disease Center, Agricultural Research Service, USDA (Pugh, Tabatabai, Bricker, Phillips, Zehr, Belzer), PO Box 70, Ames, IA 50010, and Department of Zoology, Iowa State University (Mayfield), Ames, IA 50011.

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Marshall Phillips From the National Animal Disease Center, Agricultural Research Service, USDA (Pugh, Tabatabai, Bricker, Phillips, Zehr, Belzer), PO Box 70, Ames, IA 50010, and Department of Zoology, Iowa State University (Mayfield), Ames, IA 50011.

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Emilie S. Zehr From the National Animal Disease Center, Agricultural Research Service, USDA (Pugh, Tabatabai, Bricker, Phillips, Zehr, Belzer), PO Box 70, Ames, IA 50010, and Department of Zoology, Iowa State University (Mayfield), Ames, IA 50011.

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Carol A. Belzer From the National Animal Disease Center, Agricultural Research Service, USDA (Pugh, Tabatabai, Bricker, Phillips, Zehr, Belzer), PO Box 70, Ames, IA 50010, and Department of Zoology, Iowa State University (Mayfield), Ames, IA 50011.

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SUMMARY

A study was conducted to determine whether subcomponent proteins (previously identified as BCSP20, BCSP31, and BCSP45, and the corresponding recombinant proteins rBCSP20, rBCSP31, and rBCSP45) that were recovered from the cell surface of Brucella abortus strain 19 were immunogenic and protective for mice when compared with Brucella cell surface protein (bcsp) and with a proteinase K-treated lipopolysaccharide (pklps) extracted from B abortus strain 2308. Protection was evaluated after challenge exposure with a virulent culture of B abortus strain 2308, using CD-I or BALB/c mice or both inoculated with vaccines of various combinations and concentrations, with and without pklps or bcsp. Protection was assessed by enumeration of splenic colony-forming units, reduced mean splenic weight relative to controls, and the relative serologic responses (immune response) in an elisa.

The general results indicate that bcsp, pklps, bcsp20, and bcsp31 are immunogenic or protective or both. Protectiveness was not observed for each of the recombinant proteins; however, results from the combined recombinant protein vaccine study suggest the immunogenicity of the recombinant proteins. The apparent immune-inducing properties of bcsp20 and bcsp31 are thought to be attributable to the presence of an immunogenic and protective bcsp fraction (possibly lipopolysaccharide) still associated. Serologic results support our conclusion that each of the recombinant protein vaccines did not induce a protective response comparable to that of bcsp or pklps, even when the subcomponents were combined.

Although the results suggest that the subcomponents of bcsp apparently induced partial protection, they are thought to be only a part of the antigens contained in bcsp that influence the serologic response. Our findings may serve as an experimental model to determine the mechanisms involved in the protective responses induced by Brucella antigens.

SUMMARY

A study was conducted to determine whether subcomponent proteins (previously identified as BCSP20, BCSP31, and BCSP45, and the corresponding recombinant proteins rBCSP20, rBCSP31, and rBCSP45) that were recovered from the cell surface of Brucella abortus strain 19 were immunogenic and protective for mice when compared with Brucella cell surface protein (bcsp) and with a proteinase K-treated lipopolysaccharide (pklps) extracted from B abortus strain 2308. Protection was evaluated after challenge exposure with a virulent culture of B abortus strain 2308, using CD-I or BALB/c mice or both inoculated with vaccines of various combinations and concentrations, with and without pklps or bcsp. Protection was assessed by enumeration of splenic colony-forming units, reduced mean splenic weight relative to controls, and the relative serologic responses (immune response) in an elisa.

The general results indicate that bcsp, pklps, bcsp20, and bcsp31 are immunogenic or protective or both. Protectiveness was not observed for each of the recombinant proteins; however, results from the combined recombinant protein vaccine study suggest the immunogenicity of the recombinant proteins. The apparent immune-inducing properties of bcsp20 and bcsp31 are thought to be attributable to the presence of an immunogenic and protective bcsp fraction (possibly lipopolysaccharide) still associated. Serologic results support our conclusion that each of the recombinant protein vaccines did not induce a protective response comparable to that of bcsp or pklps, even when the subcomponents were combined.

Although the results suggest that the subcomponents of bcsp apparently induced partial protection, they are thought to be only a part of the antigens contained in bcsp that influence the serologic response. Our findings may serve as an experimental model to determine the mechanisms involved in the protective responses induced by Brucella antigens.

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