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Modulation of Caspase Activation by Toxoplasma gondii

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Host-Pathogen Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 470))

Abstract

Apoptosis plays crucial roles for the outcome of infection with various infectious agents. The host’s apoptotic program may be modulated after infection in order to combat the pathogen or to restrict the immune response. In addition, distinct microorganisms alter the apoptotic program of the host in order to meet the requirements for their further distribution. The activation of caspases (i.e., cysteine proteases with specificity for aspartic acid residues) preludes the disassembly of the cell in response to apoptosis-inducing stimuli. This depends on the proteolytic cleavage of inactive proforms into catalytically active subunits. Analyses of the proteolysis and the enzymatic activity of caspases therefore represent valuable tools to study apoptotic programs during infection. The apicomplexan parasite Toxoplasma gondii interferes with the caspase cascade of its host cell in order to facilitate intracellular survival. The modulation of caspase activation by T. gondii is determined by SDS-PAGE and immunoblotting with caspase-specific antibodies. Furthermore, the impact of the parasite on caspase activity is fluorimetrically determined by measuring the cleavage of caspase-specific substrate analogues.

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References

  1. Liles, W. C. (1997) Apoptosis – role in infection and inflammation. Curr. Opin. Infect. Dis. 10, 165–170.

    Article  Google Scholar 

  2. Lüder C. G. K., Gross, U. and Lopes, M. F. (2001) Intracellular protozoan parasites and apoptosis: diverse strategies to modulate parasite-host interactions. Trends Parasitol. 17, 480–486.

    Article  PubMed  Google Scholar 

  3. Opferman, J. T. and Korsmeyer, S. J. (2003) Apoptosis in the development and maintenance of the immune system. Nature Immunol. 4, 410–415.

    Article  CAS  Google Scholar 

  4. Williams, G. T. (1994) Programmed cell death: a fundamental protective response to pathogens. Trends Microbiol. 2, 463–464.

    Article  CAS  PubMed  Google Scholar 

  5. Hengartner, M. O. (2000) The biochemistry of apoptosis. Nature 407, 770–776.

    Article  CAS  PubMed  Google Scholar 

  6. Earnshaw, W. C., Martins, L. M. and Kaufmann, S. H. (1999) Mammalian caspases: structure, activation, substrates, and functions during apoptosis. Ann. Rev. Biochem. 68, 383–424.

    Article  CAS  PubMed  Google Scholar 

  7. Krammer, P. H. (2000) CD95’s deadly mission in the immune system. Nature 407, 789–795.

    Article  CAS  PubMed  Google Scholar 

  8. Danial, N. N. and Korsmeyer, S. J. (2004) Cell death: critical control points. Cell 116, 205–219.

    Article  CAS  PubMed  Google Scholar 

  9. Green, D. R. and Reed, J. C. (1998) Mitochondria and apoptosis. Science 281, 1309–1312.

    Article  CAS  PubMed  Google Scholar 

  10. Anderson, P. (1997) Kinase cascades regulating entry into apoptosis. Microbiol. Mol. Biol. Rev. 61, 33–46.

    CAS  PubMed  Google Scholar 

  11. Li, P., Nijhawan, D., Budihardjo, I., Srinivasula, S. M., Ahmad, M., Alnemri, E. S. and Wang, X. (1997) Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91, 479–489.

    Article  CAS  PubMed  Google Scholar 

  12. Schaumburg, F., Hippe, D., Vutova, P. and Lüder, C. G. K. (2006) Pro- and antiapoptotic activities of protozoan parasites. Parasitology 132, S69–S85.

    Article  CAS  PubMed  Google Scholar 

  13. Lüder, C. G. K. and Gross, U. (2005) Apoptosis and its modulation during infection with Toxoplasma gondii: Molecular mechanisms and role in pathogenesis. In: Griffin, D. E, ed. Role of apoptosis in infection. Curr. Topics Microbiol. Immunol. 289, 219–238.

    Google Scholar 

  14. Goebel, S., Gross, U. and Lüder, C. G. K. (2001) Inhibition of host cell apoptosis by Toxoplasma gondii is accompanied by reduced activation of the caspase cascade and alterations of poly(ADP-ribose) polymerase expression. J. Cell Sci. 114, 3495–3505.

    CAS  PubMed  Google Scholar 

  15. Vutova, P., Wirth, M., Hippe, D., Gross, U., Schulze-Osthoff, K., Schmitz, I. and Lüder, C. G. K. (2007) Toxoplasma gondii inhibits Fas/CD95-triggered cell death by inducing aberrant processing and degradation of caspase 8. Cell. Microbiol. 9, 1556–1570.

    Article  CAS  PubMed  Google Scholar 

  16. Scaffidi, C., Fulda, S., Srinivasan, A., Friesen, C., Li, F., Tomaselli, K. J., Debatin, K.-M., Krammer, P. H. and Peter, M. E. (1998) Two CD95 (APO-1/Fas) signaling pathways. EMBO J. 17, 1675–1687.

    Article  CAS  PubMed  Google Scholar 

  17. Ferrari, D., Stepczynska, A., Los, M., Wesselborg, S. and Schulze-Osthoff, K. (1998) Differential regulation and ATP requirement for caspase-8 and caspase-3 activation during CD95- and anticancer drug-induced apoptosis. J. Exp. Med. 188, 979–984.

    Article  CAS  PubMed  Google Scholar 

  18. Riedl, S. J. and Shi, Y. (2004) Molecular mechanisms of caspase regulation during apoptosis. Nature Rev. Mol. Cell Biol. 5, 897–907.

    Article  CAS  Google Scholar 

  19. Gross, U., Müller, W. A., Knapp, S. and Heesemann, J. (1991) Identification of a virulence-associated antigen of Toxoplasma gondii by use of a mouse monoclonal antibody. Infect. Immun. 59, 4511–4516.

    CAS  PubMed  Google Scholar 

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Hippe, D., Gais, A., Gross, U., Lüder, C.G. (2009). Modulation of Caspase Activation by Toxoplasma gondii . In: Rupp, S., Sohn, K. (eds) Host-Pathogen Interactions. Methods in Molecular Biology, vol 470. Humana Press. https://doi.org/10.1007/978-1-59745-204-5_19

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  • DOI: https://doi.org/10.1007/978-1-59745-204-5_19

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-886-7

  • Online ISBN: 978-1-59745-204-5

  • eBook Packages: Springer Protocols

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