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Infection and Immunity, April 2004, p. 2081-2087, Vol. 72, No. 4
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.4.2081-2087.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Intraspleen Delivery of a DNA Vaccine Coding for Superoxide Dismutase (SOD) of Brucella abortus Induces SOD-Specific CD4+ and CD8+ T Cells

Carola Muñoz-Montesino,1 Edilia Andrews,1 Rodolfo Rivers,1 Andrés González-Smith,1 Gustavo Moraga-Cid,1 Hugo Folch,2 Sandra Céspedes,1 and Angel A. Oñate1*

Molecular Immunology Laboratory, Department of Microbiology, Faculty of Biological Sciences, Universidad de Concepción, Concepción,1 Department of Immunology, Faculty of Medicine, Universidad Austral, Valdivia, Chile2

Received 26 September 2003/ Returned for modification 28 October 2003/ Accepted 7 January 2004

In the development of vaccines capable of providing immunity against brucellosis, Cu-Zn superoxide dismutase (SOD) has been demonstrated to be one of the protective immunogens of Brucella abortus. In an earlier study, we provided strong evidence that intramuscular injection with a plasmid DNA carrying the SOD gene (pcDNA-SOD) was able to induce a protective immune response. The present study was designed to characterize T-cell immune responses after an intraspleen (i.s.) vaccination of BALB/c mice with pcDNA-SOD. Animals vaccinated with pcDNA-SOD did not develop SOD-specific antibodies, at least until week 4 after immunization (the end of the experiment), and in vitro stimulation of their splenocytes with either recombinant Cu-Zn SOD or crude Brucella protein induced the secretion of gamma interferon (IFN-{gamma}), but not interleukin-4, and elicited the induction of cytotoxic-T-lymphocyte activity. Upon analyzing the SOD-specific T-cell responses, the pcDNA-SOD vaccination was found to be stimulating both CD4+- and CD8+-T-cell populations. However, only the CD4+ population was able to produce IFN-{gamma} and only the CD8+ population was able to induce cytotoxic activity. Nevertheless, although i.s. route vaccination induces a significant level of protection in BALB/c mice against challenge with the virulent B. abortus strain 2308, vaccination by the intramuscular route with a similar amount of plasmid DNA does not protect. Based on these results, we conclude that i.s. immunization with pcDNA-SOD vaccine efficiently induced a Th1 type of immune response and a protective response that could be related to IFN-{gamma} production and cytotoxic activity against infected cells by SOD-specific CD4+ and CD8+ T cells, respectively.

* Corresponding author. Mailing address: Department of Microbiology, Faculty of Biological Sciences, Universidad de Concepción, P.O. Box 152-C, Concepción, Chile. Phone: (056)-41-204118. Fax: (056)-41-245975. E-mail: aonate{at}udec.cl.

Editor: J. B. Bliska

Infection and Immunity, April 2004, p. 2081-2087, Vol. 72, No. 4
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.4.2081-2087.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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