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Clinical and Vaccine Immunology, July 2009, p. 1025-1032, Vol. 16, No. 7
1071-412X/09/$08.00+0 doi:10.1128/CVI.00067-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Development of a Murine Mycobacterial Growth Inhibition Assay for Evaluating Vaccines against Mycobacterium tuberculosis
,
Marcela Parra,1
Amy L. Yang,1
JaeHyun Lim,1
Kristopher Kolibab,1
Steven Derrick,1
Nathalie Cadieux,1
Liyanage P. Perera,2
William R. Jacobs,3
Michael Brennan,1,4 and
Sheldon L. Morris1*
Center for Biologics Evaluation and Research, United States Food and Drug Administration, Bethesda, Maryland,1 Metabolism Branch, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland,2 Howard Hughes Medical Institute, Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York,3 Aeras Global TB Vaccine Foundation, Rockville, Maryland4
Received 6 February 2009/ Returned for modification 7 March 2009/ Accepted 11 May 2009
The development and characterization of new tuberculosis (TB) vaccines has been impeded by the lack of reproducible and reliable in vitro assays for measuring vaccine activity. In this study, we developed a murine in vitro mycobacterial growth inhibition assay for evaluating TB vaccines that directly assesses the capacity of immune splenocytes to control the growth of Mycobacterium tuberculosis within infected macrophages. Using this in vitro assay, protective immune responses induced by immunization with five different types of TB vaccine preparations (Mycobacterium bovis BCG, an attenuated M. tuberculosis mutant strain, a DNA vaccine, a modified vaccinia virus strain Ankara [MVA] construct expressing four TB antigens, and a TB fusion protein formulated in adjuvant) can be detected. Importantly, the levels of vaccine-induced mycobacterial growth-inhibitory responses seen in vitro after 1 week of coculture correlated with the protective immune responses detected in vivo at 28 days postchallenge in a mouse model of pulmonary tuberculosis. In addition, similar patterns of cytokine expression were evoked at day 7 of the in vitro culture by immune splenocytes taken from animals immunized with the different TB vaccines. Among the consistently upregulated cytokines detected in the immune cocultures are gamma interferon, growth differentiation factor 15, interleukin-21 (IL-21), IL-27, and tumor necrosis factor alpha. Overall, we have developed an in vitro functional assay that may be useful for screening and comparing new TB vaccine preparations, investigating vaccine-induced protective mechanisms, and assessing manufacturing issues, including product potency and stability.
* Corresponding author. Mailing address: FDA/CBER, Building 29/Room 502, 29 Lincoln Drive, Bethesda, MD 20892. Phone: (301) 496-5978. Fax: (301) 435-5675. E-mail: sheldon.morris{at}fda.hhs.gov
Published ahead of print on 20 May 2009.
Supplemental material for this article may be found at http://cvi.asm.org/.
Clinical and Vaccine Immunology, July 2009, p. 1025-1032, Vol. 16, No. 7
1071-412X/09/$08.00+0 doi:10.1128/CVI.00067-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Copyright © 2009 by the American Society for Microbiology. For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»