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Infect Immun, August 1998, p. 3952-3958, Vol. 66, No. 8
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Regulation of Macrophage Gene Expression by Mycobacterium tuberculosis: Down-Regulation of Mitochondrial Cytochrome c Oxidase

Silvia Ragno, Iris Estrada-Garcia,dagger Robert Butler, and M. Joseph Colston*

Division of Mycobacterial Research, National Institute for Medical Research, London NW7 1AA, United Kingdom

Received 18 March 1997/Returned for modification 3 June 1997/Accepted 30 April 1998

We have investigated changes in gene expression in mouse peritoneal macrophages following infection with virulent Mycobacterium tuberculosis. Using differential-display reverse transcription-PCR (RT-PCR), we have identified a gene that was markedly down-regulated within 6 h of infection and remained so for the duration of the experiment (5 days). On sequencing, this gene was found to encode the murine cytochrome c oxidase subunit VIIc (COX VIIc). Down-regulation of COX VIIc during M. tuberculosis infection was confirmed by three independent techniques: limiting-dilution RT-PCR, RNase protection assay, and Northern analysis. Limiting-dilution RT-PCR and Northern analysis were also used to analyze the specificity of this regulation; heat-killed M. tuberculosis, Mycobacterium bovis BCG, and latex beads had no effect on expression of COX VIIc. Down-regulation of this enzyme was also confirmed by using adherent cells isolated from spleens of M. tuberculosis-infected mice. These ex vivo macrophages showed apoptotic features, suggesting a possible involvement of cytochrome c oxidase in the programmed cell death of the host cells.

* Corresponding author. Mailing address: Division of Mycobacterial Research, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, United Kingdom. Phone: (0181) 959 3666. Fax: (0181) 913 8528. E-mail: j-colsto{at}nimr.mrc.ac.uk.

dagger Present address: Instituto Politecnico Nacional, Departmento de Immunologia, Mexico DF 11340, Mexico.

Infect Immun, August 1998, p. 3952-3958, Vol. 66, No. 8
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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