Phagocytosis of Candida albicans by RNAi-Treated Drosophila S2 Cells

  • Shannon L. Stroschein-Stevenson
  • Edan Foley
  • Patrick H. O’Farrell
  • Alexander D. Johnson
Part of the Methods in Molecular Biology book series (MIMB, volume 470)

Abstract

Phagocytosis is a highly conserved aspect of innate immunity. Drosophila melanogaster has an innate immune system with many similarities to that of mammals and has been used to successfully model many aspects of innate immunity. The recent availability of Ribo Nucleic Acid interference (RNAi) libraries for Drosophila has made it possible to efficiently screen for genes important in aspects of innate immunity. We have screened an RNAi library representing 7216 fly genes conserved among metazoans to identify proteins required for the phagocytosis of the human fungal pathogen Candida albicans.

Key words

Candida albicans Drosophila S2 cells cell culture RNAi double-stranded RNA phagocytosis immunofluorescence 

References

  1. 1.
    Elrod-Erickson, M., Mishra, S., and Schneider, D. (2000) Interactions between the cellular and humoral immune responses in Drosophila. Curr Biol 10, 781–784.Google Scholar
  2. 2.
    Hoffmann, J. A., Kafatos, F. C., Janeway, C. A., and Ezekowitz, R. A. (1999) Phylogenetic perspectives in innate immunity. Science 284, 1313–1318.CrossRefPubMedGoogle Scholar
  3. 3.
    Romani, L. (2004) Immunity to fungal infections. Nat Rev Immunol 4, 1–23.CrossRefPubMedGoogle Scholar
  4. 4.
    Alarco, A. M., Marcil, A., Chen, J., Suter, B., Thomas, D., and Whiteway, M. (2004) Immune-deficient Drosophila melanogaster: a model for the innate immune response to human fungal pathogens. J Immunol 172, 5622–5628.PubMedGoogle Scholar
  5. 5.
    Cheng, L. W., and Portnoy, D. A. (2003) Drosophila S2 cells: an alternative infection model for Listeria monocytogenes. Cell Microbiol 5, 875–885.CrossRefPubMedGoogle Scholar
  6. 6.
    Dionne, M. S., Ghori, N., and Schneider, D. S. (2003) Drosophila melanogaster is a genetically tractable model host for Mycobacterium marinum. Infect Immun 71, 3540–3550.CrossRefPubMedGoogle Scholar
  7. 7.
    Mansfield, B. E., Dionne, M. S., Schneider, D. S., and Freitag, N. E. (2003) Exploration of host-pathogen interactions using Listeria monocytogenes and Drosophila melanogaster. Cell Microbiol 5, 901–911.CrossRefPubMedGoogle Scholar
  8. 8.
    Schneider, D., and Shahabuddin, M. (2000) Malaria parasite development in a Drosophila model. Science 288, 2376–2379.CrossRefPubMedGoogle Scholar
  9. 9.
    Lavine, M. D., and Strand, M. R. (2002) Insect hemocytes and their role in immunity. Insect Biochem Mol Biol 32, 1295–1309.CrossRefPubMedGoogle Scholar
  10. 10.
    Agaisse, H., Burrack, L. S., Philips, J. A., Rubin, E. J., Perrimon, N., and Higgins, D. E. (2005) Genome-wide RNAi screen for host factors required for intracellular bacterial infection. Science 309, 1248–1251.CrossRefPubMedGoogle Scholar
  11. 11.
    Philips, J. A., Rubin, E. J., and Perrimon, N. (2005) Drosophila RNAi screen reveals CD36 family member required for mycobacterial infection. Science 309, 1251–1253.CrossRefPubMedGoogle Scholar
  12. 12.
    Ramet, M., Manfruelli, P., Pearson, A., Mathey-Prevot, B., and Ezekowitz, R. A. (2002) Functional genomic analysis of phagocytosis and identification of a Drosophila receptor for E. coli. Nature 416, 644–648.Google Scholar
  13. 13.
    Stroschein-Stevenson, S. L., Foley, E., O’Farrell, P. H., and Johnson, A. D. (2006) Identification of Drosophila gene products required for phagocytosis of Candida albicans. PLoS Biol 4, e4.CrossRefPubMedGoogle Scholar
  14. 14.
    Edmond, M. B., Wallace, S. E., McClish, D. K., Pfaller, M. A., Jones, R. N., and Wenzel, R. P. (1999) Nosocomial bloodstream infections in United States hospitals: a three-year analysis. Clin Infect Dis 29, 239–244.CrossRefPubMedGoogle Scholar
  15. 15.
    Calderone, R. (Ed.) (2002) Host recognition by Candida species, ASM Press, ASM Press, Washington, DC.Google Scholar
  16. 16.
    Foley, E., and O’Farrell, P. H. (2004) Functional dissection of an innate immune response by a genome-wide RNAi screen. PLoS Biol 2, E203.CrossRefPubMedGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Shannon L. Stroschein-Stevenson
    • 1
  • Edan Foley
    • 2
  • Patrick H. O’Farrell
    • 3
  • Alexander D. Johnson
    • 4
  1. 1.Department of Microbiology and ImmunologyUniversity of California, San FranciscoSan FranciscoUSA
  2. 2.Department of Medical Microbiology and ImmunologyUniversity of AlbertaEdmontonCanada
  3. 3.Department of Biochemistry and BiophysicsUniversity of California, San FranciscoSan FranciscoUSA
  4. 4.Department of Microbiology and Immunology, and Department of Biochemistry and BiophysicsUniversity of California, San FranciscoSan FranciscoUSA

Personalised recommendations