Macrophages are the first line of defense against Mycobacterium tuberculosis and other microorganisms that circumvent the mechanical defenses of the airways. When macrophages encounter particles, they ingest them in a portion of the plasma membrane and form an intracellular vesicle termed a phagosome. Simultaneous with ingestion, signal transduction pathways are triggered that activate the microbicidal systems of the macrophage. Most bacteria are killed when phagosome membranes fuse with lysosome membranes and lysosome contents gain access to the ingested bacteria. In contrast, M. tuberculosis survives in macrophages within phagosomes that do not fuse with lysosomes, and thereby avoid being killed. The overall goal of the proposed research is to determine the molecular mechanism whereby M tuberculosis avoids phagosome-lysosome fusion. We will test the hypothesis that M. tuberculosis specifically blocks a step in the signal transduction pathway that culminates in Ca2+-dependent phagosome-lysosome fusion. As increases in intracellular ionized calcium concentrations ([Ca2+]i) are proven to be essential for phagosome-lysosome fusion in human neutrophils, we will determine whether increases in [Ca2+]i occur when macrophages ingest M. tuberculosis. If not increases in [Ca2+]i occur, we will identify the most proximal step in the signal transduction pathway that is defective and will characterize the mechanism of inhibition of this step in detail. If increases in [Ca2+]i do occur when macrophages ingest M. tuberculosis, we will determine whether Ca2+- dependent effectors (annexins and calmodulin-dependent protein kinase II) are functional. If we find signal transduction is not directly affected by Mr. tuberculosis, we will test the alternative hypothesis that M. tuberculosis enters macrophages by a receptor-mediated pathway that does not normally culminate in phagosome-lysosome fusion. If this proves to be the case, we will characterize the receptor(s) involved. We will also characterize the mechanism of ingestion, to determine whether it more closely resembles phagocytosis or receptor-mediated endocytosis. The information obtained from the proposed studies will guide efforts to prevent persistent infection with M. tuberculosis by significantly enhancing our understanding of the microbe-macrophage interaction.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL051992-01
Application #
3370547
Study Section
Special Emphasis Panel (ZHL1-CSR-N (S3))
Project Start
1993-09-30
Project End
1996-08-31
Budget Start
1993-09-30
Budget End
1994-08-31
Support Year
1
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of California San Francisco
Department
Type
Schools of Medicine
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Peters, Wendy; Ernst, Joel D (2003) Mechanisms of cell recruitment in the immune response to Mycobacterium tuberculosis. Microbes Infect 5:151-8
Nagabhushanam, Vijaya; Solache, Alejandra; Ting, Li-Min et al. (2003) Innate inhibition of adaptive immunity: Mycobacterium tuberculosis-induced IL-6 inhibits macrophage responses to IFN-gamma. J Immunol 171:4750-7
Kincaid, Eleanor Z; Ernst, Joel D (2003) Mycobacterium tuberculosis exerts gene-selective inhibition of transcriptional responses to IFN-gamma without inhibiting STAT1 function. J Immunol 171:2042-9
Peters, W; Scott, H M; Chambers, H F et al. (2001) Chemokine receptor 2 serves an early and essential role in resistance to Mycobacterium tuberculosis. Proc Natl Acad Sci U S A 98:7958-63
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Ernst, J D (2000) Bacterial inhibition of phagocytosis. Cell Microbiol 2:379-86
Ting, L M; Kim, A C; Cattamanchi, A et al. (1999) Mycobacterium tuberculosis inhibits IFN-gamma transcriptional responses without inhibiting activation of STAT1. J Immunol 163:3898-906
Majeed, M; Perskvist, N; Ernst, J D et al. (1998) Roles of calcium and annexins in phagocytosis and elimination of an attenuated strain of Mycobacterium tuberculosis in human neutrophils. Microb Pathog 24:309-20
Ernst, J D (1998) Macrophage receptors for Mycobacterium tuberculosis. Infect Immun 66:1277-81
Larsson, M; Majeed, M; Ernst, J D et al. (1997) Role of annexins in endocytosis of antigens in immature human dendritic cells. Immunology 92:501-11

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