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• W621278369 abstract "Persistence in human macrophages is central to the virulence of Mycobacteriumtuberculosis, which is the causative agent of tuberculosis. Although theintracellular parasitism is apparent, molecular determinants of mycobacterialvirulence are not well understood.The current investigation identified virulent genes of M. tuberculosis bymeasuring survivability of Mycobacterium smegmatis recombinants inside ahuman monocytic cell line THP-1 after acquiring various virulent gene candidatesof M. tuberculosis. These gene candidates included nine virulent genecandidates suggested by other studies, five genomic polymorphisms identified inhypervirulent strains of M. tuberculosis using microarray-based comparativegenomic hybridization, and ten single nucleotide polymorphisms identified in thehypervirulent strains using full genome sequencing. Interestingly, onlyrecombinants harboring a truncated Rv2820c and a known virulent gene mce1Asurvived significantly better than vector control after six hours of ex vivoinfection.As nucleotide sequencing indicated that the truncated Rv2820c loses around 60%of gene at 3’ end, ex vivo survivability of M. smegmatis recombinants harboringthe last 60% of Rv2820c as well as the intact Rv2820c was measured, but wassimilar to that of vector control. The 3’ truncated portion itself did not altermycobacterial survivability ex vivo, but its presence did compromise the survivaladvantage gained due to the truncated Rv2820c.To determine whether the truncated and the intact Rv2820c could enhancemycobacterial virulence in vivo, these two alleles were transformed intoMycobacterium marinum and their recombinants were used to infect zebrafish.In vivo infection showed that zebrafish infected with the recombinant harboringtruncated Rv2820c died significantly faster than vector control, whereas therecombinant harboring intact Rv2820c behaved similarly to vector control.Results indicated that the truncated Rv2820c, but not the intact Rv2820c, couldenhance mycobacterial virulence both ex vivo and in vivo.Additional nucleotide sequencing revealed that the 3’ truncation in Rv2820c iscaused by a Beijing/W-defining deletion RD207 and is commonly found inBeijing/W strains of M. tuberculosis. Non-Beijing/W strains possess the intactRv2820c conversely. Since Beijing/W strains have proven to be more virulentthan non-Beijing/W strains both ex vivo and in vivo, the truncated Rv2820c maybe one of the Beijing/W-specific virulence determinants.To confirm that Rv2820c of Beijing/W strains really enhances M. tuberculosissurvival in human macrophages, the truncated Rv2820c was transformed intonon-Beijing/W M. tuberculosis strains and their recombinants were used to infectTHP-1 cells. Ex vivo infection confirmed that the truncated Rv2820c couldenhance M. tuberculosis survival inside human macrophages, but is unlikely toinduce a different profile of cytokine secretion from infected macrophages.In conclusion, the current study demonstrated that the truncated Rv2820c ofBeijing/W strains could enhance mycobacterial virulence both ex vivo and in vivo.Enhanced phenotypic virulence, however, was not observed for the intactRv2820c of non-Beijing/W strains. The truncated Rv2820c may be one of theBeijing/W-specific virulence determinants and collaboratively contribute to thehigh phenotypic virulence of this family." @default.
• W621278369 created "2016-06-24" @default.
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• W621278369 date "2015-05-12" @default.
• W621278369 modified "2023-09-25" @default.
• W621278369 title "The study of virulence determinants of mycobacterium tuberculosis" @default.
• W621278369 doi "https://doi.org/10.5353/th_b4784975" @default.
• W621278369 hasPublicationYear "2015" @default.
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