This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kawahara, M.
Right arrow Articles by Honda, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kawahara, M.
Right arrow Articles by Honda, M.

 Previous Article  |  Next Article 

Infection and Immunity, December 2002, p. 6614-6620, Vol. 70, No. 12
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.12.6614-6620.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Dynamics of Gamma Interferon, Interleukin-12 (IL-12), IL-10, and Transforming Growth Factor ß mRNA Expression in Primary Mycobacterium bovis BCG Infection in Guinea Pigs Measured by a Real-Time Fluorogenic Reverse Transcription-PCR Assay

Mamoru Kawahara,1,2 Tadashi Nakasone,1,3 and Mitsuo Honda1,3*

National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640,1 Japanese Foundation of AIDS Prevention, Minato-ku, Tokyo 105-0001,2 Japan Science and Technology Corporation, Kawaguchi-shi, Saitama 332-0012, Japan3

Received 22 April 2002/ Returned for modification 19 June 2002/ Accepted 21 August 2002

The guinea pig has been utilized as a model for studying infectious diseases because its reactions closely resemble those of humans biologically and immunologically. However, the cytokine responses in this animal remain to be studied. Initially, we established a quantitative assay using a real-time reverse transcription-PCR (RT-PCR) to measure guinea pig gamma interferon (IFN-{gamma}), interleukin-12 (IL-12), IL-10, and transforming growth factor ß (TGF-ß) mRNA. By preparing primer-fluorogenic probe sets for these cytokines and standard RNA templates corresponding to the target sequence of each cytokine, we obtained linear standard curves essential for quantitative determination. In guinea pigs immunized by intradermal (i.d.) vaccination with the Tokyo strain of Mycobacterium bovis BCG (0.1 mg) or else hyperimmunized with the same vaccine (10 mg) given intravenously (i.v.), peripheral blood mononuclear cells (PBMCs) at 4 weeks showed an increase in IFN-{gamma} mRNA expression in the latter but not the former animals. However, at week 10, IFN-{gamma} mRNA expression was markedly elevated in PBMCs, spleen cells, and cells in bronchoalveolar lavage fluid in both the i.d.- and the i.v.-immunized animals, the level of expression being 10 times higher in the latter. In contrast, the expression levels of IL-12 mRNA in PBMCs, spleen cells, and BAL cells were not enhanced in either group at 10 weeks postimmunization. The expression of IL-10 and TGF-ß increased slightly only in PBMCs. Regardless of differences in the levels of cytokine responses, the magnitudes of the purified protein derivative of tuberculin-specific delayed-type hypersensitivity (DTH) skin reactions for the two groups did not differ significantly at 8 weeks postvaccination. In this study, we quantitatively measured IL-10, IL-12, TGF-ß, and IFN-{gamma} mRNA in BCG-immunized guinea pigs and showed that the level of IFN-{gamma} mRNA expression does not necessarily reflect the magnitude of the DTH response, suggesting that there may be an intricate relationship between protective immunity, the level of IFN-{gamma}, and the DTH response. Thus, our quantitative assay would be of use for the development of vaccines using guinea pig models.

* Corresponding author. Mailing address: National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan. Phone: 81 3 5285 1111. Fax: 81 3 5285 1150. E-mail: mhonda{at}nih.go.jp.

Editor: J. D. Clements

Infection and Immunity, December 2002, p. 6614-6620, Vol. 70, No. 12
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.12.6614-6620.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Wang, B., Henao-Tamayo, M., Harton, M., Ordway, D., Shanley, C., Basaraba, R. J., Orme, I. M. (2007). A Toll-Like Receptor-2-Directed Fusion Protein Vaccine against Tuberculosis. CVI 14: 902-906 [Abstract] [Full Text]  
  • Tree, J. A., Elmore, M. J., Javed, S., Williams, A., Marsh, P. D. (2006). Development of a Guinea Pig Immune Response-Related Microarray and Its Use To Define the Host Response following Mycobacterium bovis BCG Vaccination. Infect. Immun. 74: 1436-1441 [Abstract] [Full Text]  
  • Izumi, Y., Ami, Y., Matsuo, K., Someya, K., Sata, T., Yamamoto, N., Honda, M. (2003). Intravenous Inoculation of Replication-Deficient Recombinant Vaccinia Virus DIs Expressing Simian Immunodeficiency Virus Gag Controls Highly Pathogenic Simian-Human Immunodeficiency Virus in Monkeys. J. Virol. 77: 13248-13256 [Abstract] [Full Text]  
  • Fuller, C. L., Flynn, J. L., Reinhart, T. A. (2003). In Situ Study of Abundant Expression of Proinflammatory Chemokines and Cytokines in Pulmonary Granulomas That Develop in Cynomolgus Macaques Experimentally Infected with Mycobacterium tuberculosis. Infect. Immun. 71: 7023-7034 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»