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Clinical and Vaccine Immunology, August 2009, p. 1121-1126, Vol. 16, No. 8
1071-412X/09/$08.00+0     doi:10.1128/CVI.00112-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Recombinant Mycobacterium bovis BCG Expressing the Chimeric Protein of Antigen 85B and ESAT-6 Enhances the Th1 Cell-Mediated Response{triangledown}

Ying Xu,1,2 Wei Liu,1 Hongbo Shen,1 Jingran Yan,1 Di Qu,2 and Honghai Wang1*

State Key Laboratory of Genetic Engineering, School of Life Science, Fudan University, Shanghai, People's Republic of China,1 Key Laboratory of Medical Molecular Virology, Shanghai Medical College of Fudan University, Shanghai, People's Republic of China2

Received 11 March 2009/ Returned for modification 1 April 2009/ Accepted 1 June 2009

The chimeric protein that relies on the T-cell epitopes of antigen 85B (Ag85B) and the 6-kDa early secreted antigen target (ESAT-6) has been demonstrated to augment the Th1 immune response. In this study, we developed a recombinant Mycobacterium bovis BCG (rBCG) strain that secretes the chimeric protein of Ag85B and ESAT-6 (rBCG-AN-E-AC). Immunization with this rBCG strain induced stronger antigen-specific gamma interferon (IFN-{gamma}) activities, as determined by an enzyme-linked immunospot assay, and higher levels of antigen-specific CD4+ and CD8+ T-cell responses than those in the control groups immunized with either rBCG expressing the Ag85B-ESAT-6 fusion protein (rBCG-A-E) or BCG. Likewise, rBCG-AN-E-AC significantly increased the level of production of the major Th1 cytokines IFN-{gamma} and tumor necrosis factor alpha in splenocyte cultures to levels comparable to those elicited by control BCG. Moreover, the antigen-specific immunoglobulin 2c (IgG2c)/IgG1 ratio for mice immunized with rBCG-AN-E-AC was also much higher than the ratios for the other immunized groups. Together, these results indicate that this rBCG-AN-E-AC strain enhances the Th1 cell-mediated response and may serve as a potential vaccine against M. tuberculosis.

* Corresponding author. Mailing address: State Key Laboratory of Genetic Engineering, School of Life Science, Fudan University, No. 220 Handan Road, Shanghai 200433, People's Republic of China. Phone: 86-21-65643777. Fax: 86-21-65648376. E-mail: hhwang{at}fudan.edu.cn

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

Clinical and Vaccine Immunology, August 2009, p. 1121-1126, Vol. 16, No. 8
1071-412X/09/$08.00+0     doi:10.1128/CVI.00112-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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