Rapid and spontaneous loss of phthiocerol dimycocerosate (PDIM) from Mycobacterium tuberculosis grown in vitro: implications for virulence studies
Pilar Domenech,Michael B. Reed +1 more
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TLDR
It is revealed that H37Rv is highly prone to losing the ability to synthesize the cell wall lipid phthiocerol dimycocerosate (PDIM) during extended periods of in vitro culture, raising a very serious issue in regard to the interpretation of putative virulence factors.Abstract:
Isolated in vitro more than half a century ago, the H37Rv strain of Mycobacterium tuberculosis still remains the strain of choice for the majority of laboratories conducting in vivo studies of TB pathogenesis. In this report we reveal that H37Rv is highly prone to losing the ability to synthesize the cell wall lipid phthiocerol dimycocerosate (PDIM) during extended periods of in vitro culture. In addition, H37Rv stocks that have been held in vitro for even a short length of time should be thought of as a heterogeneous population of PDIM-positive and PDIM-negative cell types. We demonstrate that after weekly subculture of PDIM-positive isolates over a period of 20 weeks, the proportion of PDIM-negative cells rises above 30 %. That PDIM biosynthesis is negatively selected in vitro is evident from the broad range of mutation types we observe within cultures originating from a single PDIM-positive parental clone. Moreover, the appearance of these multiple mutation types coupled with an enhanced growth rate of PDIM-negative bacteria ensures that 'PDIM-less' clones rapidly dominate in vitro cultures. It has been known for almost a decade that strains of M. tuberculosis that lack PDIM are severely attenuated during in vivo infection. Therefore, the loss of PDIM raises a very serious issue in regard to the interpretation of putative virulence factors where heterogeneous parental cultures are potentially being compared in vivo to recombinant clones isolated within a PDIM-negative background. It is essential that researchers undertaking in vivo virulence studies confirm the presence of PDIM within all recombinant clones and the parental strains they are derived from.read more
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Virulence factors of the Mycobacterium tuberculosis complex
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Gluconeogenic carbon flow of tricarboxylic acid cycle intermediates is critical for Mycobacterium tuberculosis to establish and maintain infection
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Variation among Genome Sequences of H37Rv Strains of Mycobacterium tuberculosis from Multiple Laboratories
Thomas R. Ioerger,Yicheng Feng,Krishna Ganesula,Xiaohua Chen,Karen M. Dobos,Sarah M. Fortune,William R. Jacobs,Valerie Mizrahi,Tanya Parish,Eric J. Rubin,Christopher M. Sassetti,James C. Sacchettini +11 more
TL;DR: This genome-wide catalog of genetic differences can help explain any phenotypic differences that might be found, including a frameshift mutation in the mycocerosic acid synthase gene which causes two of the strains to be deficient in biosynthesis of the surface glycolipid phthiocerol dimycoceroate (PDIM).
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References
More filters
Journal ArticleDOI
Gene Expression Omnibus: NCBI gene expression and hybridization array data repository
TL;DR: The Gene Expression Omnibus (GEO) project was initiated in response to the growing demand for a public repository for high-throughput gene expression data and provides a flexible and open design that facilitates submission, storage and retrieval of heterogeneous data sets from high-power gene expression and genomic hybridization experiments.
Journal ArticleDOI
Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence
Stewart T. Cole,Roland Brosch,Julian Parkhill,Thierry Garnier,Carol Churcher,David Harris,Stephen V. Gordon,Karin Eiglmeier,S. Gas,Clifton E. Barry,Fredj Tekaia,K. Badcock,D. Basham,D. Brown,Tracey Chillingworth,R. Connor,Robert L. Davies,K. Devlin,Theresa Feltwell,S. Gentles,N. Hamlin,S. Holroyd,T. Hornsby,Kay Jagels,Anders Krogh,J. McLean,Sharon Moule,Lee Murphy,K. Oliver,J. Osborne,Michael A. Quail,Marie-Adèle Rajandream,Jane Rogers,S. Rutter,K. Seeger,Jason Skelton,Rob Squares,S. Squares,John Sulston,K. Taylor,Sally Whitehead,Bart Barrell +41 more
TL;DR: The complete genome sequence of the best-characterized strain of Mycobacterium tuberculosis, H37Rv, has been determined and analysed in order to improve the understanding of the biology of this slow-growing pathogen and to help the conception of new prophylactic and therapeutic interventions.
Journal ArticleDOI
Genes required for mycobacterial growth defined by high density mutagenesis
TL;DR: The use of transposon site hybridization (TraSH) is described to comprehensively identify the genes required by the causative agent, Mycobacterium tuberculosis, for optimal growth, suggesting that the minimal gene set required for survival varies greatly between organisms with different evolutionary histories.
Journal ArticleDOI
New use of BCG for recombinant vaccines.
Charles K. Stover,V F de la Cruz,Thomas R. Fuerst,Jeanne E. Burlein,L. A. Benson,L. T. Bennett,Geetha P. Bansal,J. F. Young,Mong Hong Lee,Graham F. Hatfull,Scott B. Snapper,Raúl G. Barletta,William R. Jacobs,Barry R. Bloom +13 more
TL;DR: Extrachromosomal and integrative expression vectors carrying the regulatory sequences for major BCG heat-shock proteins have been developed and can elicit long-lasting humoral and cellular immune responses to foreign antigens in mice.
Global tuberculosis control: surveillance planning financing. WHO report 2003.
TL;DR: In this paper, the authors argue that if the current rate of DOTS expansion in maintained the 70% detection target will not be reached by 2005, if that target is ever to be reached DOTS programmes must improve case finding within designated DOTS areas and must expand to new areas.
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