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Flow cytometric detection of host cell apoptosis induced by bacterial infection

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Methods in Cell Science

Abstract

We detail two methods for detection of cell death induced by infection of a human monocytic cell line with invasive Campylobacter bacteria. Staining with a natural ligand for exposed phosphatidylserine residues coupled with propidum iodide discriminated between apoptosis and necrosis. Additionally, cells infected with a bacterial strain expressing green fluorescent protein stained with dye sensitive to mitochondrial membrane potential demonstrated a direct association of bacteria with dying cells. Analyses of cells stained by these methods employing flow cytometry enumerated proportions of cell populations undergoing either apoptosis or necrosis after bacterial infection in vitro.

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References

  1. Bedner E, Burfeind P, Hsieh TC, Wu JM, Aguero-Rosenfeld ME, Melamed MR, Horowitz HW, Wormser GP, Darzynkiewicz Z (1998). Cell cycle effects and induction of apoptosis caused by infection of HL-60 cells with human granulocytic ehrli-plots. Proportion of cells in each quadrant is shown above each panel. chiosis pathogen measured by flow and laser scanning cytometry. Cytom 33: 47-55.

    Google Scholar 

  2. Berninghausen O, Leippe M (1997). Necrosis versus apoptosis as the mechanism of target cell death induced by Entamoeba histolytica. Infect Immun 65: 3615-3621.

    Google Scholar 

  3. Collazo CM, Galán JE (1997). The invasion-associated type III system of Salmonella typhimurium directs the translocation of Sip proteins into the host cell. Mol Micro 24: 747-756.

    Google Scholar 

  4. Darzynkiewicz Z, Li X, Gong J (1994). Analysis of Cell viability: Discrimination of cells dying by apoptosis. In: Darzynkiewicz Z, Robinson JP, Crissman HA (eds), Methods in cell biology. New York: Academic Press 41: 16-39.

    Google Scholar 

  5. Finlay BB, Cossart P (1997). Exploitation of mammalian host cell functions by bacterial pathogens. Science 276: 718-725.

    Google Scholar 

  6. Hobbie S, Chen LM, Davis RJ, Galán JE (1997). Involvement of mitogen-activated protein kinase pathways in the nuclear responses and cytokine production induced by Salmonella typhimurium in cultured intestinal epithelial cells. J Immunol 159: 5550-5559.

    Google Scholar 

  7. Konkel ME, Kim BJ, Rivera-amill V, Garvis SG (1999). Bacterial secreted proteins are required for the internalization of Campylobacter jejuni into cultured mammalian cells. Mol Micro 32: 691-701.

    Google Scholar 

  8. Konkel ME, Joens LA, Mixter PF (2000). Molecular characterization of Campylobacter jejuni virulence determinants. In: Nachamkin I, Blaser MJ (eds), Campylobacter jejuni, 2nd ed. Washington DC: ASM Press, pp. 217-240.

    Google Scholar 

  9. Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, van Oers MHJ (1994). Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84: 1415-1420.

    Google Scholar 

  10. Kroemer G, Dallaporta B, Rishe-Rignon M (1998). The mitochondrial death/life regulator in apoptosis and necrosis Ann Rev Physiol 60: 619-642.

    Google Scholar 

  11. Lee CA (1997). Type III secretion systems: machines to deliver bacterial proteins into eukaryotic cells? Trends Micro 5: 148-156.

    Google Scholar 

  12. Nicholson DW, Ali A, Thornberry NA, Vaillancourt JP, Ding CK, Gallant M, Gareau Y, Griffin PR, Labelle M, Lazebnik YA, Munday NA, Raju SM, Smulson ME, Ting-Ting Y, Yu VL, Miller DK (1995). Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature 376: 37-43.

    Google Scholar 

  13. Rosqvist R, Hâkansson S, Fosberg A, Wolf-Katz H (1995). Functional conservation of the secretion and translocation machinery for virulence proteins of yersiniae, salmonellae, and shigellae. EMBO J 14: 4187-4195.

    Google Scholar 

  14. Schwartz LM, Osborne BA (1993). Programmed cell death, apoptosis and killer genes. Imm Today 14: 582-590.

    Google Scholar 

  15. Sgnoc R, Boeck G, Dietrich H, Gruber J, Wick G (1994). Simultaneous determination of cell surface antigens and apoptosis. Trends Gen 10: 41-42.

    Google Scholar 

  16. Tsuchiya S, Kobayashi Y, Goto Y, Okumura H, Nakae S, Konno T, Tada K (1982). Induction of maturation in cultured human monocytic leukemia cells by a phorbol diester. Cancer Res 42: 1530-1536.

    Google Scholar 

  17. Wyllie AH (1980). Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284: 555-556.

    Google Scholar 

  18. Zychlinsky A, Prevost MC, Sansonetti PJ (1992). Shigella flexneri induces apoptosis in infected macrophages. Nature 358: 167-169.

    Google Scholar 

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Authors
  1. Michael E. Konkel
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  2. Philip F. Mixter
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Konkel, M.E., Mixter, P.F. Flow cytometric detection of host cell apoptosis induced by bacterial infection. Methods Cell Sci 22, 209–215 (2000). https://doi.org/10.1023/A:1009804427679

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