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Clinical and Vaccine Immunology, August 2009, p. 1139-1145, Vol. 16, No. 8
1071-412X/09/$08.00+0     doi:10.1128/CVI.00424-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

T-Cell mRNA Expression in Response to Mycobacterium bovis BCG Vaccination and Mycobacterium bovis Infection of White-Tailed Deer{triangledown}

Tyler C. Thacker,* Mitchell V. Palmer, and W. Ray Waters

U.S. Department of Agriculture, Agricultural Research Service, Bacterial Diseases of Livestock Research Unit, National Animal Disease Center, 2300 Dayton Ave., Ames, Iowa 50010

Received 15 November 2008/ Returned for modification 10 December 2008/ Accepted 1 June 2009

Understanding immune responses of white-tailed deer (WTD) to infection with Mycobacterium bovis provides insight into mechanisms of pathogen control and may provide clues to development of effective vaccine strategies. WTD were vaccinated with either M. bovis BCG strain Pasteur or BCG strain Danish. Both vaccinees and unvaccinated controls were subsequently inoculated with virulent M. bovis via the intratonsillar route. Real-time PCR was used to assess T-cell mRNA expression in peripheral blood leukocytes (PBL) from animals following vaccination and infection. Recall T-cell responses were measured by assessing the relative expression of gamma interferon (IFN-{gamma}), T-cell-specific T-box transcription factor (Tbet), interleukin 12p40 (IL-12p40), IL-12p35, IL-23p19, FoxP3, IL-17, and GATA3 in PBL stimulated in vitro with purified protein derivative (PPD) of M. bovis or a recombinant fusion protein, ESAT6-CFP10. Animals vaccinated with BCG Danish expressed more IFN-{gamma} and Tbet than either BCG Pasteur-vaccinated animals or unvaccinated controls. BCG Pasteur-vaccinated animals expressed more GATA3 than either group. After infection, unvaccinated controls expressed more Tbet and IL-12p40 than vaccinated animals. BCG Pasteur-vaccinated animals expressed more GATA3 than either the unvaccinated controls or the BCG Danish-vaccinated animals after infection. Animals were divided into pathology groups to correlate gene expression with severity of pathology. Animals in the visible lesion group expressed more Tbet and IFN-{gamma} than animals that were culture negative, while Tbet and IFN-{gamma} expression in the culture-positive, no-visible-lesion group was intermediate. GATA3 expression inversely correlated with pathology. Overall, expression of immune response genes correlated more closely with pathology than vaccination treatment.

* Corresponding author. Mailing address: National Animal Disease Center, Bacterial Diseases of Livestock Research Unit, 2300 Dayton Ave., Ames, IA 50010. Phone: (515) 663-7294. Fax: (515) 663-7458. E-mail: tyler.thacker{at}ars.usda.gov

{triangledown} Published ahead of print on 10 June 2009.

Clinical and Vaccine Immunology, August 2009, p. 1139-1145, Vol. 16, No. 8
1071-412X/09/$08.00+0     doi:10.1128/CVI.00424-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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