Skip to main content
SpringerLink
Log in

Methods of Analyzing Chromatin Changes Accompanying Apoptosis of Target Cells in Killer Cell Assays

  • Protocol
Natural Killer Cell Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 121))

Abstract

Natural killer (NK) cells kill their targets by inducing two distinct modes of cell death—apoptosis and necrosis (1–4). The end result in both cases is target cell lysis, broadly defined as a loss of membrane integrity and most usually assessed by either chromium release or uptake of certain nonvital dyes such as eosin, trypan blue, or propidium iodide.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 74.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Duke R. C., Cohen J. J., and Chervenak R. (1986) Differences in target cell DNA fragmentation induced by mouse cytotoxic T cells and natural killer cells. J.Immunol. 137, 1442–1447.

    PubMed  CAS  Google Scholar 

  2. Zychlinsky A., Zheng L. M., Liu C. C., and Young J. D. E. (1991) Cytotoxic lymphocytes induce both apoptosis and necrosis in target cells.J. Immunol. 146, 393–400.

    PubMed  CAS  Google Scholar 

  3. Shi L., Kam C. M., Powers J. C., Aebersold R., and Greenberg A. H. (1992) Purification of three cytotoxic lymphocyte granule serine proteases that induce apoptosis through distinct substrate and target cell interactions. J. Exp. Med. 176, 1521–1529.

    Article  PubMed  CAS  Google Scholar 

  4. Kagi D., Ledermann B., Burki K., Seiler P., Odermatt B., Olsen K. J., Podack E. R., Zinkernagel R. M., and Hengartner H. (1994) Cytotoxicity mediated by T cells and natural killer cells is greatly impaired in perforin-deficient mice. Nature 369, 31–37.

    Article  PubMed  CAS  Google Scholar 

  5. Duke R. C., Ojcius D., and Young J. D.-E. (1996) Cell suicide in health and disease. Sci. Am. 275, 80–87.

    Article  PubMed  CAS  Google Scholar 

  6. Greenberg A. H. (1996) Granzyme B-induced apoptosis. Adv. Exp. Med. Biol. 406, 219–228.

    PubMed  CAS  Google Scholar 

  7. Arase H., Arase N., and Saito T. (1995) Fas-mediated cytotoxicity by freshly isolated natural killer cells. J. Exp. Med. 181, 1235–1238.

    Article  PubMed  CAS  Google Scholar 

  8. Eischen C. M. and Leibson P. J. (1997) Role for NK-cell-associated Fas ligand in cell-mediated cytotoxicity and apoptosis. Res. Immunol. 148, 164–169.

    Article  PubMed  CAS  Google Scholar 

  9. Duke R. C., Chervenak R., and Cohen J. J. (1983) Endogenous endonuclease-induced DNA fragmentation, an early event in cell-mediated cytolysis. Proc. Natl. Acad. Sci. USA. 80, 6351–6355.

    Article  Google Scholar 

  10. Duke R. C., Zulauf R., Nash P. B., Young J. D.-E., and Ojcius D. (1994) Cytolysis mediated by ionophores and pore-forming agents, role of intracellular calcium in apoptosis. FASEB J. 8, 237–246.

    PubMed  CAS  Google Scholar 

  11. Duke R. C., Persechini P. M., Chang S., Liu C.-C., Cohen J. J., and Young J. D.-E. (1989) Purified perforin induces target cell lysis but not DNA fragmentation. J. Exp. Med. 170, 1451–1456.

    Article  PubMed  CAS  Google Scholar 

  12. Kagi D., Ledermann B., Burki K., Zinkernagel R. M., and Hengartner H. (1996) Molecular mechanisms of lymphocyte-mediated cytotoxicity and their role in immunological protection and pathogenesis in vivo. Annu. Rev. Immunol. 14, 207–232.

    Article  PubMed  CAS  Google Scholar 

  13. Liu C. C., Persechini P. M., and Young J. D. (1995) Perforin and lymphocyte-mediated cytolysis. Imm. Rev. 146, 145–175.

    Article  CAS  Google Scholar 

  14. Wyllie A. H. (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284, 555–557.

    Article  PubMed  CAS  Google Scholar 

  15. Matzinger P. (1991) The JAM test, a simple assay for DNA fragmentation and cell death. J. Immunol. Methods 145, 185–192.

    Article  PubMed  CAS  Google Scholar 

  16. Gavrieli Y., Sherman Y., and Ben-Sasson S. A. (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. CellBiol. 119, 493–501.

    Article  CAS  Google Scholar 

  17. Gorczyca W., Gong J., and Darzynkiewicz Z. (1993) Detection of DNA strand breaks in individual apoptotic cells by the in situ terminal deoxynucleotidyl transferase and nick translation assays. Cancer Res. 53, 1945–1951.

    PubMed  CAS  Google Scholar 

  18. Rouvier E., Luiciani M. F., and Golstein P. 1993. Fas involvement in Ca(2+)-independent T cell-mediated cytotoxicity. J. Exp. Med. 177, 195–200.

    Google Scholar 

  19. Tanaka M., Suda T., Haze K., Nakamura N., Sato K., Kimura F., Motoyoshi K., Mizuki M., Tagawa S., Ohga S., Hatake K., Drummond A. H., and Nagata S. (1996) Fas ligand in human serum. Nat. Med. 2, 317–322.

    Article  PubMed  CAS  Google Scholar 

  20. Vignaux F., Vivier E., Malissen B., Depraetere V., Nagata S., and Golstein P. (1995) TCR/CD3 coupling to Fas-based cytotoxicity. J. Exp. Med. 181, 781–786.

    Article  PubMed  CAS  Google Scholar 

  21. Ojcius D. M., Zychlinski A., Zheng L. M. and Young J. D. Y. (1991) Ionophore-induced apoptosis, role of DNA fragmentation and calcium fluxes. Exp. Cell Res. 197, 43–49.

    Article  PubMed  CAS  Google Scholar 

  22. Duke R. C., Sellins K. S., and Cohen J. J. (1988) Cytotoxic lymphocyte-derived lytic granules do not induce DNA fragmentation in target cells. J. Immunol. 141, 2141–2145.

    Google Scholar 

  23. Miyawaki T., Uehara T., Nibu R., Tsuji T., Yachie A., Yonehara S., and Taniguchi N. (1992) Differential expression of apoptosis-relatedFas antigen on lymphocyte subpopulations in human peripheral blood. J. Immunol. 149, 3753–3758.

    PubMed  CAS  Google Scholar 

  24. Leithauser F., Dhein J., Mechtersheimer G., Koretz K., Bruderlein S., Henne C., Schmidt A., Debatin K. M., Krammer P. H., and Moller P. (1993) Constitutive and induced expression of APO-1, a new member of the nerve growth factor/ tumor necrosis factor receptor superfamily, in normal and neoplastic cells. Lab. Invest. 69, 415–427.

    PubMed  CAS  Google Scholar 

  25. Maciejewski J., Selleri C., Anderson S., and Young N. S. (1995) Fas antigen expression on CD34+ human marrow cells is induced by interferon gamma and tumor necrosis factor alpha and potentiates cytokine-mediated hematopoietic suppression in vitro. Blood 85, 3183–3190.

    PubMed  CAS  Google Scholar 

  26. George T., Yu Y. Y., Liu J., Davenport C., Lemieux S., Stoneman E., Mathew P. A., Kumar V., and Bennett M. (1997) Allorecognition by murine natural killer cells, lysis of T-lymphoblasts and rejection of bone-marrow grafts. Imm. Rev. 155, 29–40.

    Article  CAS  Google Scholar 

  27. Sellins K. S. and Cohen J. J. (1991) Cytotoxic T lymphocytes induce different types of DNA damage in target cells of different origins. J. Immunol. 147, 795–803.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. CERES Pharmaceuticals, Ltd, Denver, CO

    Richard C. Duke

  2. Medical Oncology, University of Colorado Health Science Center, Denver, CO

    Richard C. Duke

  3. Eleanor Roosevelt Institute for Cancer Research, Denver, CO

    Richard C. Duke

Authors
  1. Richard C. Duke
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Fox Chase Cancer Center, Philadelphia, PA

    Kerry S. Campbell

  2. Basel Institute for Immunology, Basel, Switzerland

    Marco Colonna

Rights and permissions

Reprints and permissions

Copyright information

© 1999 Humana Press Inc.

About this protocol

Cite this protocol

Duke, R.C. (1999). Methods of Analyzing Chromatin Changes Accompanying Apoptosis of Target Cells in Killer Cell Assays. In: Campbell, K.S., Colonna, M. (eds) Natural Killer Cell Protocols. Methods in Molecular Biology, vol 121. Humana Press. https://doi.org/10.1385/1-59259-044-6:125

Download citation

  • DOI: https://doi.org/10.1385/1-59259-044-6:125

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-683-3

  • Online ISBN: 978-1-59259-044-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation