title: Mycobacterium tuberculosis resides in nonacidified vacuoles in endocytically competent alveolar macrophages from patients with tuberculosis and HIV infection.
abstract: Alveolar macrophages (AM) are the first professional phagocytes encountered by aerosols containing infections in the lungs, and their phagocytic capacity may be affected by these infections or environmental particles. The aim of this study was to evaluate the innate endocytic and phagocytic properties of human AM obtained from patients with pulmonary tuberculosis and to characterize the vacuoles in which Mycobacterium tuberculosis bacilli reside in vivo. AM were obtained by bronchoalveolar lavage from patients with suspected tuberculosis and from asymptomatic volunteers (controls). Clinical case definitions were based on mycobacterial culture of respiratory specimens and HIV serology. To assess phagocytosis, endocytosis, and acidification of the endosomal system, AM were cultured with IgG-coated polystyrene beads, dextran, and a pH-sensitive reporter (3-(2,4-dinitroanilino)-3-amino-N-methyldipropylamine) and were evaluated by light and immunoelectron microscopy. Cells from 89 patients and 10 controls were studied. We found no significant difference between the two groups in the ability of AM either to ingest beads and dextran or to deliver them to acidified lysosomes. In AM from patients with tuberculosis, the bacilli were located in vacuoles that failed to accumulate endocytosed material and were not acidified. We concluded that AM from patients with tuberculosis and HIV infections were competent to endocytose and phagocytose material and to deliver the material to functional, acidified lysosomes. M. tuberculosis residing in these AM arrests the progression of their phagosomes, which fail to fuse with acidified lysosomes. This confirms, for the first time in humans with tuberculosis and HIV, the conclusions from previous animal and in vitro studies.
authors.affiliation: Malawi-Liverpool-Wellcome Trust Clinical Research Program, College of Medicine, Blantyre, Malawi.
authors.forename: Henry C
authors.initials: HC
authors.lastname: Mwandumba
authors.forename: David G
authors.initials: DG
authors.lastname: Russell
authors.forename: Mukanthu H
authors.initials: MH
authors.lastname: Nyirenda
authors.forename: Jennifer
authors.initials: J
authors.lastname: Anderson
authors.forename: Sarah A
authors.initials: SA
authors.lastname: White
authors.forename: Malcolm E
authors.initials: ME
authors.lastname: Molyneux
authors.forename: S Bertel
authors.initials: SB
authors.lastname: Squire
mesh_terms: D000328:Adult; D002478:Cells, Cultured; D003911:Dextrans / Q000378:metabolism; D004136:Dinitrobenzenes / Q000378:metabolism; D004705:Endocytosis / Q000276:immunology*; D015658:HIV Infections / Q000276:immunology* / Q000382:microbiology* / Q000473:pathology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D016676:Macrophages, Alveolar / Q000378:metabolism / Q000382:microbiology* / Q000648:ultrastructure; D009169:Mycobacterium tuberculosis / Q000378:metabolism / Q000472:pathogenicity* / Q000648:ultrastructure; D010587:Phagocytosis / Q000276:immunology; D014397:Tuberculosis, Pulmonary / Q000276:immunology* / Q000382:microbiology* / Q000473:pathology; D014617:Vacuoles / Q000378:metabolism / Q000382:microbiology* / Q000648:ultrastructure
chemical_list: D003911:Dextrans; D004136:Dinitrobenzenes; C042432:3-(2,4-dinitroanilino)-3'-amino-N-methyldipropylamine