title: The Delta fbpA mutant derived from Mycobacterium tuberculosis H37Rv has an enhanced susceptibility to intracellular antimicrobial oxidative mechanisms, undergoes limited phagosome maturation and activates macrophages and dendritic cells.
abstract: Mycobacterium tuberculosis H37Rv (Mtb) excludes phagocyte oxidase (phox) and inducible nitric oxide synthase (iNOS) while preventing lysosomal fusion in macrophages (MPhis). The antigen 85A deficient (Delta fbpA) mutant of Mtb was vaccinogenic in mice and the mechanisms of attenuation were compared with MPhis infected with H37Rv and BCG. Delta fbpA contained reduced amounts of trehalose 6, 6, dimycolate and induced minimal levels of SOCS-1 in MPhis. Blockade of oxidants enhanced the growth of Delta fbpA in MPhis that correlated with increased colocalization with phox and iNOS. Green fluorescent protein-expressing strains within MPhis or purified phagosomes were analysed for endosomal traffick with immunofluorescence and Western blot. Delta fbpA phagosomes were enriched for rab5, rab11, LAMP-1 and Hck suggesting enhanced fusion with early, recycling and late endosomes in MPhis compared with BCG or H37Rv. Delta fbpA phagosomes were thus more mature than H37Rv or BCG although, they failed to acquire rab7 and CD63 preventing lysosomal fusion. Finally, Delta fbpA infected MPhis and dendritic cells (DCs) showed an enhanced MHC-II and CD1d expression and primed immune T cells to release more IFN-gamma compared with those infected with BCG and H37Rv. Delta fbpA was thus more immunogenic in MPhis and DCs because of an enhanced susceptibility to oxidants and increased maturation.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Texas Health Sciences Center, Houston, TX, USA.
authors.forename: Muralidhar K
authors.initials: MK
authors.lastname: Katti
authors.forename: Guixiang
authors.initials: G
authors.lastname: Dai
authors.forename: Lisa Y
authors.initials: LY
authors.lastname: Armitige
authors.forename: Carlos
authors.initials: C
authors.lastname: Rivera Marrero
authors.forename: Sundarsingh
authors.initials: S
authors.lastname: Daniel
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Singh
authors.forename: Devin R
authors.initials: DR
authors.lastname: Lindsey
authors.forename: Subramanian
authors.initials: S
authors.lastname: Dhandayuthapani
authors.forename: Robert L
authors.initials: RL
authors.lastname: Hunter
authors.forename: Chinnaswamy
authors.initials: C
authors.lastname: Jagannath
mesh_terms: D000217:Acyltransferases / Q000235:genetics*; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics*; D018949:Antigens, CD1 / Q000378:metabolism; D055610:Antigens, CD1d; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000378:metabolism* / Q000382:microbiology; D011992:Endosomes / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D007371:Interferon-gamma / Q000096:biosynthesis; D051907:Lysosomal Membrane Proteins / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism* / Q000382:microbiology; D051379:Mice; D008810:Mice, Inbred C57BL; D009169:Mycobacterium tuberculosis / Q000472:pathogenicity / Q000502:physiology*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism*; D010588:Phagosomes / Q000378:metabolism*; D017354:Point Mutation; D051572:Proto-Oncogene Proteins c-hck / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction; D000071222:Suppressor of Cytokine Signaling 1 Protein; D050826:Suppressor of Cytokine Signaling Proteins / Q000378:metabolism*; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D020079:Terminal Repeat Sequences / Q000235:genetics*; D014376:Tuberculosis / Q000276:immunology / Q000382:microbiology; D020696:rab5 GTP-Binding Proteins / Q000378:metabolism; D052118:Lysosomal-Associated Membrane Protein 1
chemical_list: D000217:Acyltransferases; D000942:Antigens, Bacterial; D018949:Antigens, CD1; D055610:Antigens, CD1d; D019204:GTP-Binding Proteins; D000949:Histocompatibility Antigens Class II; D007371:Interferon-gamma; D051907:Lysosomal Membrane Proteins; D052247:Nitric Oxide Synthase Type II; D010088:Oxidoreductases; D051572:Proto-Oncogene Proteins c-hck; D017382:Reactive Oxygen Species; D000071222:Suppressor of Cytokine Signaling 1 Protein; D050826:Suppressor of Cytokine Signaling Proteins; D020696:rab5 GTP-Binding Proteins; C052255:Lamp1 protein, mouse; D052118:Lysosomal-Associated Membrane Protein 1; C491590:Socs1 protein, mouse; C112580:antigen 85A, Mycobacterium tuberculosis; C496632:Hck protein, mouse