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Infection and Immunity, May 2006, p. 2734-2741, Vol. 74, No. 5
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.5.2734-2741.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Protective Immunity Elicited by a Divalent DNA Vaccine Encoding Both the L7/L12 and Omp16 Genes of Brucella abortus in BALB/c Mice

Deyan Luo,1,{dagger} Bing Ni,2,{dagger} Peng Li,1 Wei Shi,1 Songle Zhang,1 Yue Han,1 Liwei Mao,2 Yangdong He,2 Yuzhang Wu,2* and Xiliang Wang1*

State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China,1 Institute of Immunology PLA, Third Military Medical University, Chongqing 400038, China2

Received 14 October 2005/ Returned for modification 28 November 2005/ Accepted 1 February 2006

This study was designed to evaluate the immunogenicity and the protective efficacy of a divalent fusion DNA vaccine encoding both the Brucella abortus L7/L12 protein (ribosomal protein) and Omp16 protein (outer membrane lipoprotein), designated pcDNA3.1-L7/L12-Omp16. Intramuscular injection of this divalent DNA vaccine into BALB/c mice elicited markedly both humoral and cellular immune responses. The specific antibodies exhibited a dominance of immunoglobulin G2a (IgG2a) over IgG1. In addition, the dual-gene DNA vaccine elicited a strong T-cell proliferative response and induced a large amount of gamma interferon-producing T cells upon restimulation in vitro with recombinant fusion protein L7/L12-Omp16, suggesting the induction of a typical T-helper-1-dominated immune response in vivo. This divalent DNA vaccine could also induce a significant level of protection against challenge with the virulent strain B. abortus 544 in BALB/c mice. Furthermore, the protection level induced by the divalent DNA vaccine was significantly higher than that induced by the univalent DNA vaccines pcDNA3.1-L7/L12 or pcDNA3.1-Omp16. Taken together, the results of this study verify for the first time that the Omp16 gene can be a candidate target for a DNA vaccine against brucellosis. Additionally, a divalent genetic vaccine based on the L7/L12 and Omp16 genes can elicit a stronger cellular immune response and better immunoprotection than the relevant univalent vaccines can.

* Corresponding author. Mailing address for Yuzhang Wu: Institute of Immunology PLA, Department of Immunology, Third Military Medical University, Chongqing 400038, China. Phone: 86 23 6875 2680. Fax: 86 23 6875 2789. Email: wuyuzhang@yahoo.com. Mailing address for Xiliang Wang: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, Beijing 100071, China. Phone and fax: 86 10 6694 8678. E-mail: xiliangw{at}yahoo.com.

Editor: J. T. Barbieri

{dagger} D.L. and B.N. contributed equally to the paper.

Infection and Immunity, May 2006, p. 2734-2741, Vol. 74, No. 5
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.5.2734-2741.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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