Journal
TUBERCULOSIS
Volume 88, Issue 6, Pages 553-559Publisher
CHURCHILL LIVINGSTONE
DOI: 10.1016/j.tube.2008.03.005
Keywords
Neutral red; Phthiocerol dimycocerosates; Phenotypic variation; Virulence instability; Population heterogeneity
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Funding
- PHS HHS [HHSN266200400091C] Funding Source: Medline
- Wellcome Trust Funding Source: Medline
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The laboratory strain H37Rv represents one of the most commonly used strains in the study of Mycobacterium tuberculosis. Despite the apparent stability of the strain, the absence of a selective pressure for virulence factors could lead to the in vitro accumulation of attenuated mutants. To assess this hypothesis, we performed a systematic analysis of individual clones isolated from subcultured M. tuberculosis H37Rv and from a non-subcultured frozen stock. First, we studied two virulence indicators: neutral red staining and content of phthiocerol dimyco-cerosates (PDIMs). We found that H37Rv formed a mixed population containing wild-type cells, as well as neutral red and PDIM mutants. Then, we compared the global gene expression of 3 isolated clones (which displayed various phenotypes) and the non-subcultured stock, by micro-array analysis. This transcriptional profiting confirmed that a significant heterogeneity existed despite, and in addition to, the neutral red and PDIM phenotypes. These results strongly suggest that great caution must be taken in extrapolating data obtained with M. tuberculosis H37Rv grown in vitro, and it would be prudent to study several independent clones to obtain valid conclusions. For this purpose, the neutral red and PDIM phenotypes might be useful indicators of undesired heterogeneity. (C) 2008 Elsevier Ltd. All rights reserved.
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