IAI Accepts, published online ahead of print on 3 November 2008

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Infect. Immun. doi:10.1128/IAI.01151-08
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Immunization with recombinant Brucella spp. outer membrane proteins Omp16 or Omp19 in adjuvant induces specific CD4⁺ and CD8⁺ T cells as well as systemic and oral protection against Brucella abortus infection

Karina A. Pasquevich, Silvia M. Estein, Clara García Samartino, Astrid Zwerdling, Lorena M. Coria, Paula Barrionuevo, Carlos A. Fossati, Guillermo H. Giambartolomei, and Juliana Cassataro^*

Laboratorio de Inmunogenética, Hospital de Clínicas "José de San Martín", Facultad de Medicina. Universidad de Buenos Aires (UBA). Buenos Aires. Argentina; Instituto de Estudios de la Inmunidad Humoral (IDEHU-CONICET), Facultad de Farmacia y Bioquímica, UBA, Buenos Aires, Argentina; Laboratorio de Inmunología, Departamento de Sanidad Animal y Medicina Preventiva, Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires, Tandil, Argentina

^* To whom correspondence should be addressed. Email: jucassat{at}ffyb.uba.ar.

Available vaccines against Brucella spp. are live attenuated Brucella strains. In order to engineer a better vaccine to be used in animals and humans, in our laboratory we aim to develop an innocuous subunit vaccine. Particularly, we are interested in the outer membrane proteins (Omps) of B. abortus: Omp16 and Omp19. In this study, we assessed the use of these proteins as vaccines against Brucella in BALB/c mice. Immunization with lipidated (L)-Omp16 or L-Omp19 in incomplete Freund's adjuvant (IFA) conferred significant protection against B. abortus infection. Vaccination with unlipidated (U)-Omp16 or U-Omp19 in IFA induced a higher degree of protection than the respective lipidated versions. Moreover, the level of protection induced after U-Omp16 or U-Omp19 immunization in IFA was similar to the elicited by live B. abortus strain 19 (S19) immunization. Flow cytometric analysis showed that immunization with U-Omp16 or U-Omp19 induced antigen-specific CD4⁺ as well as CD8⁺ T cells producing interferon-. In vivo depletion of CD4⁺ or CD8⁺ T cells in U-Omp16- or U-Omp19- plus IFA-immunized mice resulted in a loss of the elicited protection, indicating that both cell types are mediating immune protection. U-Omp16 or U-Omp19 vaccination induced a T helper 1 response, systemic protection in Alum formulation and oral protection with cholera toxin adjuvant against B. abortus infection. Both immunization routes exhibited a similar degree of protection to attenuated Brucella vaccines (S19 and RB51 respectively). Overall these results indicate that U-Omp16 or U-Omp19 would be useful candidates for a subunit vaccine against human and animal brucellosis.