4.4 Article

Identification of Virulence Markers of Mycobacterium abscessus for Intracellular Replication in Phagocytes

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出版社

JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/57766

关键词

Immunology and Infection; Issue 139; Microbiology; amoeba; Acanthamoeba castellanii; Mycobacterium abscessus; transposon library; co-culture amoeba-mycobacteria; RNA extraction of intracellular mycobacteria; RNAseq

资金

  1. French Patient Association for Cystic Fibrosis Vaincre la Mucoviscidose [RF20150501377]
  2. French Patient Association for Cystic Fibrosis L'Association Gregory Lemarchal [RF20150501377]
  3. National Agency for Research (ANR program DIMIVYR) [ANR-13-BSV3-0007-01]
  4. Region Ile-de-France (Domaine d'Interet Majeur Maladies Infectieuses et Emergentes)
  5. Agence Nationale de la Recherche (ANR) [ANR-13-BSV3-0007] Funding Source: Agence Nationale de la Recherche (ANR)

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What differentiates Mycobacterium abscessus from other saprophytic mycobacteria is the ability to resist phagocytosis by human macrophages and the ability to multiply inside such cells. These virulence traits render M. abscessus pathogenic, especially in vulnerable hosts with underlying structural lung disease, such as cystic fibrosis, bronchiectasis or tuberculosis. How patients become infected with M. abscessus remains unclear. Unlike many mycobacteria, M. abscessus is not found in the environment but might reside inside amoebae, environmental phagocytes that represent a potential reservoir for M. abscessus. Indeed, M. abscessus is resistant to amoebal phagocytosis and the intra-amoeba life seems to increase M. abscessus virulence in an experimental model of infection. However, little is known about M. abscessus virulence in itself. To decipher the genes conferring an advantage to M. abscessus intracellular life, a screening of a M. abscessus transposon mutant library was developed. In parallel, a method of RNA extraction from intracellular Mycobacteria after co-culture with amoebae was developed. This method was validated and allowed the sequencing of whole M. abscessus transcriptomes inside the cells; providing, for the first time, a global view on M. abscessus adaptation to intracellular life. Both approaches give us an insight into M. abscessus virulence factors that enable M. abscessus to colonize the airways in humans.

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