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Journal of Virology, July 2005, p. 8716-8723, Vol. 79, No. 14
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.14.8716-8723.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Mycobacterial Codon Optimization Enhances Antigen Expression and Virus-Specific Immune Responses in Recombinant Mycobacterium bovis Bacille Calmette-Guérin Expressing Human Immunodeficiency Virus Type 1 Gag{dagger}

Masaru Kanekiyo,1,2 Kazuhiro Matsuo,1 Makiko Hamatake,1 Takaichi Hamano,1 Takeaki Ohsu,1 Sohkichi Matsumoto,3 Takeshi Yamada,4 Shudo Yamazaki,1 Atsuhiko Hasegawa,2 Naoki Yamamoto,1 and Mitsuo Honda1*

AIDS Research Center, National Institute of Infectious Diseases, Shinjuku, Tokyo 162-8640, Japan,1 Department of Pathobiology, School of Veterinary Medicine, Nihon University, Fujisawa, Kanagawa 252-8510, Japan,2 Department of Host Defense, Graduate School of Medicine, Osaka City University, Osaka, Osaka 545-8585, Japan,3 Department of Bacteriology, School of Dentistry, Nagasaki University, Nagasaki, Nagasaki 852-8588, Japan4

Received 22 November 2004/ Accepted 7 April 2005

Although its potential for vaccine development is already known, the introduction of recombinant human immunodeficiency virus (HIV) genes to Mycobacterium bovis bacille Calmette-Guérin (BCG) has thus far elicited only limited responses. In order to improve the expression levels, we optimized the codon usage of the HIV type 1 (HIV-1) p24 antigen gene of gag (p24 gag) and established a codon-optimized recombinant BCG (rBCG)-p24 Gag which expressed a 40-fold-higher level of p24 Gag than did that of nonoptimized rBCG-p24 Gag. Inoculation of mice with the codon-optimized rBCG-p24 Gag elicited effective immunity, as evidenced by virus-specific lymphocyte proliferation, gamma interferon ELISPOT cell induction, and antibody production. In contrast, inoculation of animals with the nonoptimized rBCG-p24 Gag induced only low levels of immune responses. Furthermore, a dose as small as 0.01 mg of the codon-optimized rBCG per animal proved capable of eliciting immune responses, suggesting that even low doses of a codon-optimized rBCG-based vaccine could effectively elicit HIV-1-specific immune responses.

* Corresponding author. Mailing address: AIDS Research Center, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan. Phone: 81-3-5285-1111. Fax: 81-3-5285-1183. E-mail: mhonda{at}nih.go.jp.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, July 2005, p. 8716-8723, Vol. 79, No. 14
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.14.8716-8723.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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