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Infection and Immunity, March 2002, p. 1245-1253, Vol. 70, No. 3
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.3.1245-1253.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Effect of Mycobacterium bovis BCG Vaccination on Interleukin-1ß and RANTES mRNA Expression in Guinea Pig Cells Exposed to Attenuated and Virulent Mycobacteria

Amminikutty Jeevan,1* Teizo Yoshimura,2 Gregory Foster,1 and David N. McMurray1

Department of Medical Microbiology and Immunology, Texas A&M University System Health Science Center, College Station, Texas 77843-1114,1 Laboratory of Molecular Immunoregulation, NCI-Frederick, Frederick, Maryland 217022

Received 12 July 2001/ Returned for modification 15 August 2001/ Accepted 16 November 2001

The effect of Mycobacterium bovis BCG vaccination on interleukin-1ß (IL-1ß) or regulated-upon-activation, normally T-cell-expressed and -secreted chemokine (RANTES) mRNA expression in guinea pig spleen cells stimulated with concanavalin A, lipopolysaccharide (LPS), phorbol myristate acetate (PMA) plus ionomycin, or purified protein derivative (PPD) was studied in vitro. Similarly, peritoneal exudate cell-derived macrophages from naïve and BCG-vaccinated guinea pigs were infected with M. bovis BCG, Mycobacterium avium, the attenuated Mycobacterium tuberculosis H37Ra strain, or virulent strains H37Rv and Erdman of M. tuberculosis. Total RNA was subjected to Northern blot analysis using probes generated from guinea pig IL-1ß or RANTES cDNA. Although IL-1ß and RANTES mRNA could be detected in the spleen cells from naïve animals stimulated with LPS or PMA plus ionomycin, the levels were significantly enhanced after BCG vaccination. mRNA expression was also elevated in macrophages infected with live mycobacteria after BCG vaccination. However, macrophages infected with the virulent H37Rv strain of M. tuberculosis showed 75 to 90% reductions in IL-1ß expression and 25 to 60% reductions in RANTES mRNA expression compared with macrophages infected with the attenuated H37Ra strain. The IL-1ß mRNA levels peaked as soon as 1 h after PPD stimulation and 4 h after M. tuberculosis H37Rv infection of macrophages. In contrast, RANTES mRNA expression was delayed until 48 h after infection. These results indicate that molecular mediators produced in response to various stimuli associated with protective immunity against mycobacteria are upregulated after BCG vaccination; however, a significantly weaker response was observed with virulent M. tuberculosis. These initial studies indicate that BCG vaccination has a positive effect on IL-1ß and RANTES mRNA expression by host cells in a highly relevant animal tuberculosis model.

* 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) 862-2436. Fax: (979) 845-3479. E-mail: ajeevan{at}medicine.tamu.edu.

Infection and Immunity, March 2002, p. 1245-1253, Vol. 70, No. 3
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.3.1245-1253.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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