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Infection and Immunity, October 2002, p. 5471-5478, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5471-5478.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Mycobacterium bovis BCG Vaccination Augments Interleukin-8 mRNA Expression and Protein Production in Guinea Pig Alveolar Macrophages Infected with Mycobacterium tuberculosis

Mark J. Lyons,1 Teizo Yoshimura,2 and David N. McMurray1*

Department of Medical Microbiology and Immunology, Texas A&M University System Health Science Center, College Station, Texas 77843,1 Laboratory of Molecular Immunoregulation, NCI—Frederick, Frederick, Maryland 217022

Received 12 March 2002/ Returned for modification 26 April 2002/ Accepted 15 July 2002

Alveolar macrophages are likely the first cell type to encounter Mycobacterium tuberculosis in a pulmonary infection, resulting in the production of chemokines. In order to evaluate this response, alveolar macrophages harvested from nonvaccinated and Mycobacterium bovis BCG-vaccinated guinea pigs were infected in vitro with live M. tuberculosis H37Ra or H37Rv (multiplicity of infection, 1:1) or cultured with lipopolysaccharide (10 µg/ml) for 3, 12, and 24 h. Interleukin-8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1) mRNA expression was determined by real-time PCR. Culture supernatants were assayed for guinea pig IL-8 protein by using a human IL-8 enzyme-linked immunosorbent assay kit. Alveolar macrophages harvested from BCG-vaccinated guinea pigs produced significantly more mRNA and protein for IL-8 than alveolar macrophages harvested from nonvaccinated guinea pigs at 12 and 24 h poststimulation or postinfection. Infection with attenuated M. tuberculosis (H37Ra) stimulated alveolar macrophages isolated from BCG-vaccinated guinea pigs to produce significantly more IL-8 mRNA than did alveolar macrophages infected with a virulent strain (H37Rv) at 12 and 24 h postinfection. Significant MCP-1 mRNA production was also detected in stimulated or infected alveolar macrophages; however, prior vaccination did not significantly affect levels of MCP-1 mRNA. Alveolar macrophages isolated from BCG-vaccinated guinea pigs produced significantly more IL-8 mRNA and protein when stimulated for 24 h with heat-killed H37Ra, heat-killed H37Rv, and H37Rv cell wall, but not mannose-capped lipoarabinomannan (ManLAM), than did cells stimulated with media alone. These observations indicate that prior vaccination may alter very early events in the M. tuberculosis-infected lung.

* Corresponding author. Mailing address: Department of Medical Microbiology and Immunology, Texas A&M University System Health Science Center, 407 Reynolds Medical Building, College Station, TX 77843-1114. Phone: (979) 845-3679. Fax: (979) 845-3479. E-mail: dmcmurray{at}tamu.edu.

Editor: S. H. E. Kaufmann

Infection and Immunity, October 2002, p. 5471-5478, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5471-5478.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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