This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Crowle, A J Articles by May, M H Search for Related Content PubMed PubMed Citation Articles by Crowle, A J Articles by May, M H

 Previous Article  |  Next Article 

Infect Immun. 1991 May; 59(5): 1823-1831

Evidence that vesicles containing living, virulent Mycobacterium tuberculosis or Mycobacterium avium in cultured human macrophages are not acidic.

Webb-Waring Lung Institute, Department of Microbiology and Immunology, University of Colorado Health Sciences Center, Denver 80262.

ABSTRACT

Mycobacterium tuberculosis and Mycobacterium avium multiply in cultured human macrophages (MP) within membrane-enclosed vesicles. These vesicles are generally assumed to be acidic. The evidence most frequently cited for this assumption is that pyrazinamide, which requires an acid pH to be effective, is effective and streptomycin, which loses most of its activity at a low pH, is poorly effective against tubercle bacilli. This assumption was tested by using the two weak bases chloroquine and NH4Cl to raise the pH of acidic vesicles in MP experimentally infected with M. tuberculosis or M. avium. An immunocytochemical locator of acidic regions in the MP was used to monitor the association of intracellular bacilli with acidity. MP were infected with M. tuberculosis or M. avium and incubated with various combinations of the drugs and the weak bases. Replication of the bacteria in the MP was measured by culture counts. Intracellular associations of the mycobacteria with acidity were assessed by electron micrographs and by using the weak base 3-(2,4-dinitroanilino)-3'-amino-N-methyl dipropylamine, which was detected with colloidal gold-labeled antibodies. It was confirmed by immunocytochemistry that both chloroquine and NH4Cl raise the pH of acidic vesicles in the infected MP. However, neither caused any pH-related change in the antimycobacterial activities of pyrazinamide or streptomycin or of the pH-independent drug isoniazid. Immunochemical analyses showed acidity to be associated with killed but not living mycobacteria in the MP. These findings suggest that living M. tuberculosis and M. avium are located in human MP in vesicles which are not acidic.

This article has been cited by other articles:

• Rampini, S. K., Selchow, P., Keller, C., Ehlers, S., Bottger, E. C., Sander, P. (2008). LspA inactivation in Mycobacterium tuberculosis results in attenuation without affecting phagosome maturation arrest. Microbiology 154: 2991-3001 [Abstract] [Full Text]  
• Brumell, J. H., Scidmore, M. A. (2007). Manipulation of Rab GTPase Function by Intracellular Bacterial Pathogens. Microbiol. Mol. Biol. Rev. 71: 636-652 [Abstract] [Full Text]  
• Danelishvili, L., Wu, M., Stang, B., Harriff, M., Cirillo, S., Cirillo, J., Bildfell, R., Arbogast, B., Bermudez, L. E. (2007). Identification of Mycobacterium avium pathogenicity island important for macrophage and amoeba infection. Proc. Natl. Acad. Sci. USA 104: 11038-11043 [Abstract] [Full Text]  
• Vazquez, N., Greenwell-Wild, T., Rekka, S., Orenstein, J. M., Wahl, S. M. (2006). Mycobacterium avium-induced SOCS contributes to resistance to IFN-{gamma}-mediated mycobactericidal activity in human macrophages. J. Leukoc. Biol. 80: 1136-1144 [Abstract] [Full Text]  
• Chu, H., Lee, J.-H., Han, S.-H., Kim, S.-Y., Cho, N.-H., Kim, I.-S., Choi, M.-S. (2006). Exploitation of the Endocytic Pathway by Orientia tsutsugamushi in Nonprofessional Phagocytes. Infect. Immun. 74: 4246-4253 [Abstract] [Full Text]  
• Appelberg, R. (2006). Macrophage nutriprive antimicrobial mechanisms. J. Leukoc. Biol. 79: 1117-1128 [Abstract] [Full Text]  
• Newman, S. L., Gootee, L., Hilty, J., Morris, R. E. (2006). Human Macrophages Do Not Require Phagosome Acidification to Mediate Fungistatic/Fungicidal Activity against Histoplasma capsulatum. J. Immunol. 176: 1806-1813 [Abstract] [Full Text]  
• Cheadle, E. J., O'Donnell, D., Selby, P. J., Jackson, A. M. (2005). Closely Related Mycobacterial Strains Demonstrate Contrasting Levels of Efficacy as Antitumor Vaccines and Are Processed for Major Histocompatibility Complex Class I Presentation by Multiple Routes in Dendritic Cells. Infect. Immun. 73: 784-794 [Abstract] [Full Text]  
• Wagner, D., Maser, J., Lai, B., Cai, Z., Barry, C. E. III, Honer zu Bentrup, K., Russell, D. G., Bermudez, L. E. (2005). Elemental Analysis of Mycobacterium avium-, Mycobacterium tuberculosis-, and Mycobacterium smegmatis-Containing Phagosomes Indicates Pathogen-Induced Microenvironments within the Host Cell's Endosomal System. J. Immunol. 174: 1491-1500 [Abstract] [Full Text]  
• Clemens, D. L., Lee, B.-Y., Horwitz, M. A. (2004). Virulent and Avirulent Strains of Francisella tularensis Prevent Acidification and Maturation of Their Phagosomes and Escape into the Cytoplasm in Human Macrophages. Infect. Immun. 72: 3204-3217 [Abstract] [Full Text]  
• Pais, T. F., Appelberg, R. (2004). Induction of Mycobacterium avium growth restriction and inhibition of phagosome-endosome interactions during macrophage activation and apoptosis induction by picolinic acid plus IFN{gamma}. Microbiology 150: 1507-1518 [Abstract] [Full Text]  
• Mwandumba, H. C., Russell, D. G., Nyirenda, M. H., Anderson, J., White, S. A., Molyneux, M. E., Squire, S. B. (2004). Mycobacterium tuberculosis Resides in Nonacidified Vacuoles in Endocytically Competent Alveolar Macrophages from Patients with Tuberculosis and HIV Infection. J. Immunol. 172: 4592-4598 [Abstract] [Full Text]  
• Fratti, R. A., Chua, J., Deretic, V. (2003). Induction of p38 Mitogen-activated Protein Kinase Reduces Early Endosome Autoantigen 1 (EEA1) Recruitment to Phagosomal Membranes. J. Biol. Chem. 278: 46961-46967 [Abstract] [Full Text]  
• Nagabhushanam, V., Solache, A., Ting, L.-M., Escaron, C. J., Zhang, J. Y., Ernst, J. D. (2003). Innate Inhibition of Adaptive Immunity: Mycobacterium tuberculosis-Induced IL-6 Inhibits Macrophage Responses to IFN-{gamma}. J. Immunol. 171: 4750-4757 [Abstract] [Full Text]  
• Harrison, R. E., Bucci, C., Vieira, O. V., Schroer, T. A., Grinstein, S. (2003). Phagosomes Fuse with Late Endosomes and/or Lysosomes by Extension of Membrane Protrusions along Microtubules: Role of Rab7 and RILP. Mol. Cell. Biol. 23: 6494-6506 [Abstract] [Full Text]  
• Cotter, P. D., Hill, C. (2003). Surviving the Acid Test: Responses of Gram-Positive Bacteria to Low pH. Microbiol. Mol. Biol. Rev. 67: 429-453 [Abstract] [Full Text]  
• Bhattacharyya, A., Pathak, S., Basak, C., Law, S., Kundu, M., Basu, J. (2003). Execution of Macrophage Apoptosis by Mycobacterium avium through Apoptosis Signal-regulating Kinase 1/p38 Mitogen-activated Protein Kinase Signaling and Caspase 8 Activation. J. Biol. Chem. 278: 26517-26525 [Abstract] [Full Text]  
• Smith, I. (2003). Mycobacterium tuberculosis Pathogenesis and Molecular Determinants of Virulence. Clin. Microbiol. Rev. 16: 463-496 [Abstract] [Full Text]  
• Saviola, B., Woolwine, S. C., Bishai, W. R. (2003). Isolation of Acid-Inducible Genes of Mycobacterium tuberculosis with the Use of Recombinase-Based In Vivo Expression Technology. Infect. Immun. 71: 1379-1388 [Abstract] [Full Text]  
• Hou, J. Y., Graham, J. E., Clark-Curtiss, J. E. (2002). Mycobacterium avium Genes Expressed during Growth in Human Macrophages Detected by Selective Capture of Transcribed Sequences (SCOTS). Infect. Immun. 70: 3714-3726 [Abstract] [Full Text]  
• Boshoff, H. I., Mizrahi, V., Barry, C. E. III (2002). Effects of Pyrazinamide on Fatty Acid Synthesis by Whole Mycobacterial Cells and Purified Fatty Acid Synthase I. J. Bacteriol. 184: 2167-2172 [Abstract] [Full Text]  
• Webster, P. (2002). Early Intracellular Events During Internalization of Listeria monocytogenes by J774 Cells. J. Histochem. Cytochem. 50: 503-518 [Abstract] [Full Text]  
• Astarie-Dequeker, C., Carreno, S., Cougoule, C., Maridonneau-Parini, I. (2002). The protein tyrosine kinase Hck is located on lysosomal vesicles that are physically and functionally distinct from CD63-positive lysosomes in human macrophages. J. Cell Sci. 115: 81-89 [Abstract] [Full Text]  
• HUNT, J. F., ERWIN, E., PALMER, L., VAUGHAN, J., MALHOTRA, N., PLATTS-MILLS, T. A. E., GASTON, B. (2002). Expression and Activity of pH-regulatory Glutaminase in the Human Airway Epithelium. Am. J. Respir. Crit. Care Med. 165: 101-107 [Abstract] [Full Text]  
• El-Etr, S. H., Yan, L., Cirillo, J. D. (2001). Fish Monocytes as a Model for Mycobacterial Host-Pathogen Interactions. Infect. Immun. 69: 7310-7317 [Abstract] [Full Text]  
• Maurin, M., Raoult, D. (2001). Use of Aminoglycosides in Treatment of Infections Due to Intracellular Bacteria. Antimicrob. Agents Chemother. 45: 2977-2986 [Full Text]  
• Fairbairn, I. P., Stober, C. B., Kumararatne, D. S., Lammas, D. A. (2001). ATP-Mediated Killing of Intracellular Mycobacteria by Macrophages Is a P2X7-Dependent Process Inducing Bacterial Death by Phagosome-Lysosome Fusion. J. Immunol. 167: 3300-3307 [Abstract] [Full Text]  
• Stober, C. B., Lammas, D. A., Li, C. M., Kumararatne, D. S., Lightman, S. L., McArdle, C. A. (2001). ATP-Mediated Killing of Mycobacterium bovis Bacille Calmette-Guerin Within Human Macrophages Is Calcium Dependent and Associated with the Acidification of Mycobacteria-Containing Phagosomes. J. Immunol. 166: 6276-6286 [Abstract] [Full Text]  
• Neyrolles, O., Gould, K., Gares, M.-P., Brett, S., Janssen, R., O'Gaora, P., Herrmann, J.-L., Prevost, M.-C., Perret, E., Thole, J. E. R., Young, D. (2001). Lipoprotein Access to MHC Class I Presentation During Infection of Murine Macrophages with Live Mycobacteria. J. Immunol. 166: 447-457 [Abstract] [Full Text]  
• Beatty, W. L., Russell, D. G. (2000). Identification of Mycobacterial Surface Proteins Released into Subcellular Compartments of Infected Macrophages. Infect. Immun. 68: 6997-7002 [Abstract] [Full Text]  
• Ullrich, H.-J., Beatty, W. L., Russell, D. G. (2000). Interaction of Mycobacterium avium-Containing Phagosomes with the Antigen Presentation Pathway. J. Immunol. 165: 6073-6080 [Abstract] [Full Text]  
• Clemens, D. L., Lee, B.-Y., Horwitz, M. A. (2000). Mycobacterium tuberculosis and Legionella pneumophila Phagosomes Exhibit Arrested Maturation despite Acquisition of Rab7. Infect. Immun. 68: 5154-5166 [Abstract] [Full Text]  
• Garcia, R. C., Banfi, E., Pittis, M. G. (2000). Infection of Macrophage-Like THP-1 Cells with Mycobacterium avium Results in a Decrease in Their Ability to Phosphorylate Nucleolin. Infect. Immun. 68: 3121-3128 [Abstract] [Full Text]  
• Clemens, D. L., Lee, B.-Y., Horwitz, M. A. (2000). Deviant Expression of Rab5 on Phagosomes Containing the Intracellular Pathogens Mycobacterium tuberculosis and Legionella pneumophila Is Associated with Altered Phagosomal Fate. Infect. Immun. 68: 2671-2684 [Abstract] [Full Text]  
• Miller, B. H., Shinnick, T. M. (2000). Evaluation of Mycobacterium tuberculosis Genes Involved in Resistance to Killing by Human Macrophages. Infect. Immun. 68: 387-390 [Abstract] [Full Text]  
• Birkness, K. A., Swords, W. E., Huang, P.-H., White, E. H., Dezzutti, C. S., Lal, R. B., Quinn, F. D. (1999). Observed Differences in Virulence-Associated Phenotypes between a Human Clinical Isolate and a Veterinary Isolate of Mycobacterium avium. Infect. Immun. 67: 4895-4901 [Abstract] [Full Text]  
• Porte, F., Liautard, J.-P., Kohler, S. (1999). Early Acidification of Phagosomes Containing Brucella suis Is Essential for Intracellular Survival in Murine Macrophages. Infect. Immun. 67: 4041-4047 [Abstract] [Full Text]  
• Gomes, M. S., Paul, S., Moreira, A. L., Appelberg, R., Rabinovitch, M., Kaplan, G. (1999). Survival of Mycobacterium avium and Mycobacterium tuberculosis in Acidified Vacuoles of Murine Macrophages. Infect. Immun. 67: 3199-3206 [Abstract] [Full Text]  
• Berthet, F., Lagranderie, M., Gounon, P., Laurent-Winter, C., Ensergueix, D., Chavarot, P., Thouron, F., Maranghi, E., Pelicic, V., Portnoï, D., Marchal, G., Gicquel, B. (1998). Attenuation of Virulence by Disruption of the Mycobacterium tuberculosis erp Gene. Science 282: 759-762 [Abstract] [Full Text]  
• Stenger, S., Niazi, K. R., Modlin, R. L. (1998). Down-Regulation of CD1 on Antigen-Presenting Cells by Infection with Mycobacterium tuberculosis. J. Immunol. 161: 3582-3588 [Abstract] [Full Text]  
• Schaible, U. E., Sturgill-Koszycki, S., Schlesinger, P. H., Russell, D. G. (1998). Cytokine Activation Leads to Acidification and Increases Maturation of Mycobacterium avium-Containing Phagosomes in Murine Macrophages. J. Immunol. 160: 1290-1296 [Abstract] [Full Text]  
• Via, L., Fratti, R., McFalone, M, Pagan-Ramos, E, Deretic, D, Deretic, V (1998). Effects of cytokines on mycobacterial phagosome maturation. J. Cell Sci. 111: 897-905 [Abstract]  
• Nau, G. J., Guilfoile, P., Chupp, G. L., Berman, J. S., Kim, S. J., Kornfeld, H., Young, R. A. (1997). A chemoattractant cytokine associated with granulomas in tuberculosis and silicosis. Proc. Natl. Acad. Sci. USA 94: 6414-6419 [Abstract] [Full Text]  
• Via, L. E., Deretic, D., Ulmer, R. J., Hibler, N. S., Huber, L. A., Deretic, V. (1997). Arrest of Mycobacterial Phagosome Maturation Is Caused by a Block in Vesicle Fusion between Stages Controlled by rab5 and rab7. J. Biol. Chem. 272: 13326-13331 [Abstract] [Full Text]  
• Alvarez-Dominguez, C, Roberts, R, Stahl, P. (1997). Internalized Listeria monocytogenes modulates intracellular trafficking and delays maturation of the phagosome. J. Cell Sci. 110: 731-743 [Abstract]  
• Small, P., Ramakrishnan, L, Falkow, S (1994). Remodeling schemes of intracellular pathogens. Science 263: 637-639  
• Sturgill-Koszycki, S, Schlesinger, P., Chakraborty, P, Haddix, P., Collins, H., Fok, A., Allen, R., Gluck, S., Heuser, J, Russell, D. (1994). Lack of acidification in Mycobacterium phagosomes produced by exclusion of the vesicular proton-ATPase. Science 263: 678-681 [Abstract]  
• Pancholi, P, Mirza, A, Bhardwaj, N, Steinman, R. (1993). Sequestration from immune CD4+ T cells of mycobacteria growing in human macrophages. Science 260: 984-986 [Abstract]