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Infection and Immunity, July 2006, p. 4048-4057, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.01787-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Evaluation of Protection Afforded by Brucella abortus and Brucella melitensis Unmarked Deletion Mutants Exhibiting Different Rates of Clearance in BALB/c Mice

M. M. Kahl-McDonagh and T. A. Ficht*

Department of Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station, Texas 77843-4467

Received 3 November 2005/ Returned for modification 27 January 2006/ Accepted 14 April 2006

Research for novel Brucella vaccines has focused upon the development of live vaccine strains, which have proven more efficacious than killed or subunit vaccines. In an effort to develop improved vaccines, signature-tagged mutant banks were screened to identify mutants attenuated for survival. Mutants selected from these screens exhibited various degrees of attenuation characterized by the rate of clearance, ranging from a failure to grow in macrophages after 24 h of infection to a failure to persist in the mouse model beyond 8 weeks. Ideal vaccine candidates should be safe to the host, while evoking protective immunity. In the present work, we constructed unmarked deletion mutants of three gene candidates, manBA, virB2, and asp24, in both Brucella abortus and Brucella melitensis. The {Delta}asp24 mutants, which persist for extended periods in vivo, are superior to current vaccine strains and to other deletion strains tested in the mouse model against homologous challenge infection after 12, 16, and 20 weeks postvaccination. The {Delta}asp24 mutants also display superior protection compared to {Delta}manBA and {Delta}virB2 mutants against heterologous challenge in mice. From this study, a direct association between protection against infection and cytokine response was not apparent between all vaccine groups and, therefore, correlates of protective immunity will need to be considered further. A distinct correlation between persistence of the vaccine strain and protection against infection was corroborated.

* Corresponding author. Mailing address: Texas A&M University, Department of Veterinary Pathobiology, MS 4467, College Station, TX 77843-4467. Phone: (979) 845-4118. Fax: (979) 862-1088. E-mail: tficht{at}cvm.tamu.edu.

Editor: V. J. DiRita

Infection and Immunity, July 2006, p. 4048-4057, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.01787-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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