Original Article

Inhibition of Ca²⁺ Signaling by Mycobacterium tuberculosis Is Associated with Reduced Phagosome–Lysosome Fusion and Increased Survival within Human Macrophages

Correspondence to: David J. Kusner, Div. of Infectious Diseases, Dept. of Internal Medicine, University of Iowa, 200 Hawkins Dr., SW 34-I, GH, Iowa City, IA 52242. Tel:319-353-6525 Fax:319-356-4600 E-mail:david-kusner{at}uiowa.edu.

Complement receptor (CR)-mediated phagocytosis of Mycobacterium tuberculosis by macrophages results in intracellular survival, suggesting that M. tuberculosis interferes with macrophage microbicidal mechanisms. As increases in cytosolic Ca²⁺ concentration ([Ca²⁺]_c) promote phagocyte antimicrobial responses, we hypothesized that CR phagocytosis of M. tuberculosis is accompanied by altered Ca²⁺ signaling. Whereas the control complement (C)-opsonized particle zymosan (COZ) induced a 4.6-fold increase in [Ca²⁺]_c in human macrophages, no change in [Ca²⁺]_c occurred upon addition of live, C-opsonized virulent M. tuberculosis. Viability of M. tuberculosis and ingestion via CRs was required for infection of macrophages in the absence of increased [Ca²⁺]_c, as killed M. tuberculosis or antibody (Ab)-opsonized, live M. tuberculosis induced elevations in [Ca²⁺]_c similar to COZ. Increased [Ca²⁺]_c induced by Ab-opsonized bacilli was associated with a 76% reduction in intracellular survival, compared with C-opsonized M. tuberculosis. Similarly, reversible elevation of macrophage [Ca²⁺]_c with the ionophore A23187 reduced intracellular viability by 50%. Ionophore-mediated elevation of [Ca²⁺]_c promoted the maturation of phagosomes containing live C-opsonized bacilli, as evidenced by acidification and accumulation of lysosomal protein markers. These data demonstrate that M. tuberculosis inhibits CR-mediated Ca²⁺ signaling and indicate that this alteration of macrophage activation contributes to inhibition of phagosome–lysosome fusion and promotion of intracellular mycobacterial survival.

Key Words: calcium, macrophages, tuberculosis, bacterial pathogenesis, immunology

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• Dale, D. C., Boxer, L., Liles, W. C. (2008). The phagocytes: neutrophils and monocytes. Blood 112: 935-945 [Abstract] [Full Text]  
• Shi, L., Albuquerque, P. C., Lazar-Molnar, E., Wang, X., Santambrogio, L., Gacser, A., Nosanchuk, J. D. (2008). A Monoclonal Antibody to Histoplasma capsulatum Alters the Intracellular Fate of the Fungus in Murine Macrophages. Eukaryot Cell 7: 1109-1117 [Abstract] [Full Text]  
• Hinkovska-Galcheva, V., Clark, A., VanWay, S., Huang, J.-B., Hiraoka, M., Abe, A., Borofsky, M., Kunkel, R. G., Shanley, T., Shayman, J. A., Lanni, F., Petty, H. R., Boxer, L. A. (2008). Ceramide kinase promotes Ca2+ signaling near IgG-opsonized targets and enhances phagolysosomal fusion in COS-1 cells. J. Lipid Res. 49: 531-542 [Abstract] [Full Text]  
• Weiss, D. J., Souza, C. D., Evanson, O. A., Sanders, M., Rutherford, M. (2008). Bovine monocyte TLR2 receptors differentially regulate the intracellular fate of Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium. J. Leukoc. Biol. 83: 48-55 [Abstract] [Full Text]  
• Schierloh, P., Yokobori, N., Aleman, M., Landoni, V., Geffner, L., Musella, R. M., Castagnino, J., Baldini, M., Abbate, E., de la Barrera, S. S., Sasiain, M. C. (2007). Mycobacterium tuberculosis-Induced Gamma Interferon Production by Natural Killer Cells Requires Cross Talk with Antigen-Presenting Cells Involving Toll-Like Receptors 2 and 4 and the Mannose Receptor in Tuberculous Pleurisy. Infect. Immun. 75: 5325-5337 [Abstract] [Full Text]  
• Huynh, K. K., Grinstein, S. (2007). Regulation of Vacuolar pH and Its Modulation by Some Microbial Species. Microbiol. Mol. Biol. Rev. 71: 452-462 [Abstract] [Full Text]  
• Kusner, D. J., Thompson, C. R., Melrose, N. A., Pitson, S. M., Obeid, L. M., Iyer, S. S. (2007). The Localization and Activity of Sphingosine Kinase 1 Are Coordinately Regulated with Actin Cytoskeletal Dynamics in Macrophages. J. Biol. Chem. 282: 23147-23162 [Abstract] [Full Text]  
• Ferguson, J. S., Martin, J. L., Azad, A. K., McCarthy, T. R., Kang, P. B., Voelker, D. R., Crouch, E. C., Schlesinger, L. S. (2006). Surfactant Protein D Increases Fusion of Mycobacterium tuberculosis- Containing Phagosomes with Lysosomes in Human Macrophages. Infect. Immun. 74: 7005-7009 [Abstract] [Full Text]  
• Chun, J., Prince, A. (2006). Activation of Ca2+-Dependent Signaling by TLR2. J. Immunol. 177: 1330-1337 [Abstract] [Full Text]  
• Stockinger, W., Zhang, S. C., Trivedi, V., Jarzylo, L. A., Shieh, E. C., Lane, W. S., Castoreno, A. B., Nohturfft, A. (2006). Differential Requirements for Actin Polymerization, Calmodulin, and Ca2+ Define Distinct Stages of Lysosome/Phagosome Targeting. Mol. Biol. Cell 17: 1697-1710 [Abstract] [Full Text]  
• Iyer, S. S., Agrawal, R. S., Thompson, C. R., Thompson, S., Barton, J. A., Kusner, D. J. (2006). Phospholipase D1 Regulates Phagocyte Adhesion. J. Immunol. 176: 3686-3696 [Abstract] [Full Text]  
• Heung, L. J., Luberto, C., Del Poeta, M. (2006). Role of Sphingolipids in Microbial Pathogenesis. Infect. Immun. 74: 28-39 [Full Text]  
• Herrmann, T. L., Morita, C. T., Lee, K., Kusner, D. J. (2005). Calmodulin kinase II regulates the maturation and antigen presentation of human dendritic cells. J. Leukoc. Biol. 78: 1397-1407 [Abstract] [Full Text]  
• Tomioka, H., Sano, C., Sato, K., Ogasawara, K., Akaki, T., Sano, K., Cai, S. S., Shimizu, T. (2005). Combined Effects of ATP on the Therapeutic Efficacy of Antimicrobial Drug Regimens against Mycobacterium avium Complex Infection in Mice and Roles of Cytosolic Phospholipase A2-Dependent Mechanisms in the ATP-Mediated Potentiation of Antimycobacterial Host Resistance. J. Immunol. 175: 6741-6749 [Abstract] [Full Text]  
• Kang, P. B., Azad, A. K., Torrelles, J. B., Kaufman, T. M., Beharka, A., Tibesar, E., DesJardin, L. E., Schlesinger, L. S. (2005). The human macrophage mannose receptor directs Mycobacterium tuberculosis lipoarabinomannan-mediated phagosome biogenesis. J. Exp. Med. 202: 987-999 [Abstract] [Full Text]  
• de Valliere, S., Abate, G., Blazevic, A., Heuertz, R. M., Hoft, D. F. (2005). Enhancement of Innate and Cell-Mediated Immunity by Antimycobacterial Antibodies. Infect. Immun. 73: 6711-6720 [Abstract] [Full Text]  
• Fukasawa, Y., Kawamura, I., Uchiyama, R., Yamamoto, K., Kaku, T., Tominaga, T., Nomura, T., Ichiyama, S., Ezaki, T., Mitsuyama, M. (2005). Streptomycin-Dependent Exhibition of Cytokine-Inducing Activity in Streptomycin-Dependent Mycobacterium tuberculosis Strain 18b. Infect. Immun. 73: 7051-7055 [Abstract] [Full Text]  
• Thompson, C. R., Iyer, S. S., Melrose, N., VanOosten, R., Johnson, K., Pitson, S. M., Obeid, L. M., Kusner, D. J. (2005). Sphingosine Kinase 1 (SK1) Is Recruited to Nascent Phagosomes in Human Macrophages: Inhibition of SK1 Translocation by Mycobacterium tuberculosis. J. Immunol. 174: 3551-3561 [Abstract] [Full Text]  
• Vergne, I., Chua, J., Lee, H.-H., Lucas, M., Belisle, J., Deretic, V. (2005). Mechanism of phagolysosome biogenesis block by viable Mycobacterium tuberculosis. Proc. Natl. Acad. Sci. USA 102: 4033-4038 [Abstract] [Full Text]  
• Fajardo, M., Schleicher, M., Noegel, A., Bozzaro, S., Killinger, S., Heuner, K., Hacker, J., Steinert, M. (2004). Calnexin, calreticulin and cytoskeleton-associated proteins modulate uptake and growth of Legionella pneumophila in Dictyostelium discoideum. Microbiology 150: 2825-2835 [Abstract] [Full Text]  
• Iyer, S. S., Barton, J. A., Bourgoin{section}, S., Kusner, D. J. (2004). Phospholipases D1 and D2 Coordinately Regulate Macrophage Phagocytosis. J. Immunol. 173: 2615-2623 [Abstract] [Full Text]  
• Roy, D., Liston, D. R., Idone, V. J., Di, A., Nelson, D. J., Pujol, C., Bliska, J. B., Chakrabarti, S., Andrews, N. W. (2004). A Process for Controlling Intracellular Bacterial Infections Induced by Membrane Injury. Science 304: 1515-1518 [Abstract] [Full Text]  
• Vieira, O. V., Harrison, R. E., Scott, C. C., Stenmark, H., Alexander, D., Liu, J., Gruenberg, J., Schreiber, A. D., Grinstein, S. (2004). Acquisition of Hrs, an Essential Component of Phagosomal Maturation, Is Impaired by Mycobacteria. Mol. Cell. Biol. 24: 4593-4604 [Abstract] [Full Text]  
• Feng, H.-M., Whitworth, T., Olano, J. P., Popov, V. L., Walker, D. H. (2004). Fc-Dependent Polyclonal Antibodies and Antibodies to Outer Membrane Proteins A and B, but Not to Lipopolysaccharide, Protect SCID Mice against Fatal Rickettsia conorii Infection. Infect. Immun. 72: 2222-2228 [Abstract] [Full Text]  
• Hestvik, A. L. K., Hmama, Z., Av-Gay, Y. (2003). Kinome Analysis of Host Response to Mycobacterial Infection: a Novel Technique in Proteomics. Infect. Immun. 71: 5514-5522 [Abstract] [Full Text]  
• Vergne, I., Chua, J., Deretic, V. (2003). Tuberculosis Toxin Blocking Phagosome Maturation Inhibits a Novel Ca2+/Calmodulin-PI3K hVPS34 Cascade. J. Exp. Med. 198: 653-659 [Abstract] [Full Text]  
• Tobian, A. A. R., Potter, N. S., Ramachandra, L., Pai, R. K., Convery, M., Boom, W. H., Harding, C. V. (2003). Alternate Class I MHC Antigen Processing Is Inhibited by Toll-Like Receptor Signaling Pathogen-Associated Molecular Patterns: Mycobacterium tuberculosis 19-kDa Lipoprotein, CpG DNA, and Lipopolysaccharide. J. Immunol. 171: 1413-1422 [Abstract] [Full Text]  
• Lovett, J. L., Sibley, L. D. (2003). Intracellular calcium stores in Toxoplasma gondii govern invasion of host cells. J. Cell Sci. 116: 3009-3016 [Abstract] [Full Text]  
• Smith, I. (2003). Mycobacterium tuberculosis Pathogenesis and Molecular Determinants of Virulence. Clin. Microbiol. Rev. 16: 463-496 [Abstract] [Full Text]  
• Worth, R. G., Kim, M.-K., Kindzelskii, A. L., Petty, H. R., Schreiber, A. D. (2003). Signal sequence within Fcgamma RIIA controls calcium wave propagation patterns: Apparent role in phagolysosome fusion. Proc. Natl. Acad. Sci. USA 100: 4533-4538 [Abstract] [Full Text]  
• Malik, Z. A., Thompson, C. R., Hashimi, S., Porter, B., Iyer, S. S., Kusner, D. J. (2003). Cutting Edge: Mycobacterium tuberculosis Blocks Ca2+ Signaling and Phagosome Maturation in Human Macrophages Via Specific Inhibition of Sphingosine Kinase. J. Immunol. 170: 2811-2815 [Abstract] [Full Text]  
• Duan, L., Gan, H., Golan, D. E., Remold, H. G. (2002). Critical Role of Mitochondrial Damage in Determining Outcome of Macrophage Infection with Mycobacterium tuberculosis. J. Immunol. 169: 5181-5187 [Abstract] [Full Text]  
• Myers, J. T., Swanson, J. A. (2002). Calcium spikes in activated macrophages during Fc{gamma} receptor-mediated phagocytosis. J. Leukoc. Biol. 72: 677-684 [Abstract] [Full Text]  
• Russell, D. G., Mwandumba, H. C., Rhoades, E. E. (2002). Mycobacterium and the coat of many lipids. J. Cell Biol. 158: 421-426 [Abstract] [Full Text]  
• Li, J. S.-y., Chu, F., Reilly, A., Winslow, G. M. (2002). Antibodies Highly Effective in SCID Mice During Infection by the Intracellular Bacterium Ehrlichia chaffeensis Are of Picomolar Affinity and Exhibit Preferential Epitope and Isotype Utilization. J. Immunol. 169: 1419-1425 [Abstract] [Full Text]  
• Worth, R. G., Mayo-Bond, L., Kim, M.-K., van de Winkel, J. G. J., Todd, R. F. III, Petty, H. R., Schreiber, A. D. (2001). The cytoplasmic domain of Fcgamma RIIA (CD32) participates in phagolysosome formation. Blood 98: 3429-3434 [Abstract] [Full Text]  
• Kusner, D. J., Barton, J. A. (2001). ATP Stimulates Human Macrophages to Kill Intracellular Virulent Mycobacterium tuberculosis Via Calcium-Dependent Phagosome-Lysosome Fusion. J. Immunol. 167: 3308-3315 [Abstract] [Full Text]  
• Fratti, R. A., Backer, J. M., Gruenberg, J., Corvera, S., Deretic, V. (2001). Role of phosphatidylinositol 3-kinase and Rab5 effectors in phagosomal biogenesis and mycobacterial phagosome maturation arrest. J. Cell Biol. 154: 631-644 [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]  
• Rook, G.A.W., Seah, G., Ustianowski, A. (2001). M. tuberculosis: immunology and vaccination. Eur Respir J 17: 537-557 [Abstract] [Full Text]  
• Malik, Z. A., Iyer, S. S., Kusner, D. J. (2001). Mycobacterium tuberculosis Phagosomes Exhibit Altered Calmodulin-Dependent Signal Transduction: Contribution to Inhibition of Phagosome-Lysosome Fusion and Intracellular Survival in Human Macrophages. J. Immunol. 166: 3392-3401 [Abstract] [Full Text]  
• Groger, M., Holnthoner, W., Maurer, D., Lechleitner, S., Wolff, K., Mayr, B. B., Lubitz, W., Petzelbauer, P. (2000). Dermal Microvascular Endothelial Cells Express the 180-kDa Macrophage Mannose Receptor In Situ and In Vitro. J. Immunol. 165: 5428-5434 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search

TABLE OF CONTENTS