Skip to main content
SpringerLink
Log in

In Vitro Evaluation of the Apoptosis Function in Human Activated T Cells

  • Protocol
  • First Online:
CD95

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1557))

Abstract

The apoptosis function can be monitored on human lymphocytes by quantifying the induced-death upon apoptotic stimuli involving either the extrinsic or the intrinsic pathway on in vitro activated T cells. Her, we describe an in vitro assay allowing the monitoring of three different apoptosis pathways: (1) the FAS-induced pathway, (2) the activation-induced cell death (AICD), and (3) the death induced by starvation of the cells, called activated-cell autonomous death (ACAD).

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 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Anilkumar TV, Sarraf CE, Alison MR (1992) The biology and pathology of programmed cell death (apoptosis). Vet Hum Toxicol 34(3):251–254

    CAS  PubMed  Google Scholar 

  2. Aki T, Funakoshi T, Uemura K (2015) Regulated necrosis and its implications in toxicology. Toxicology 333:118–126

    Article  CAS  PubMed  Google Scholar 

  3. Worth A, Thrasher AJ, Gaspar B (2006) Autoimmune lymphoproliferative syndrome: molecular basis of disease and clinical phenotype. Br J Haematol 133:124–140

    Article  CAS  PubMed  Google Scholar 

  4. Sneller MC, Dale JK, Straus SE (2003) Autoimmune lymphoproliferative syndrome. Curr Opin Rheumatol 15:417–421

    Article  CAS  PubMed  Google Scholar 

  5. Rieux-Laucat F, Le Deist F, Fischer A (2003) Autoimmune lymphoproliferative syndromes: genetic defects of apoptosis pathways. Cell Death Differ 10:124–133

    Article  CAS  PubMed  Google Scholar 

  6. Fisher GH, Rosenberg FJ, Straus SE et al (1995) Dominant interfering Fas gene mutations impair apoptosis in a human autoimmune lymphoproliferative syndrome. Cell 81:935–946

    Article  CAS  PubMed  Google Scholar 

  7. Magerus-Chatinet A et al (2011) Onset of autoimmune lymphoproliferative syndrome (ALPS) in humans as a consequence of genetic defect accumulation. J Clin Invest 121(1):106–112

    Article  CAS  PubMed  Google Scholar 

  8. Holzelova E, Cédric Vonarbourg MS, Stolzenberg MC et al (2004) Autoimmune lymphoproliferative syndrome with somatic Fas mutations. N Engl J Med 351:1409–1418

    Article  CAS  PubMed  Google Scholar 

  9. Peter ME, Krammer PH (2003) The CD95(APO-1/Fas) DISC and beyond. Cell Death Differ 10(1):26–35

    Article  CAS  PubMed  Google Scholar 

  10. Cain K, Bratton SB, Cohen GM (2002) The Apaf-1 apoptosome: a large caspase-activating complex. Biochimie 84(2–3):203–214

    Article  CAS  PubMed  Google Scholar 

  11. Hildeman DA, Zhu Y, Mitchell TC, Kappler J, Marrack P (2002) Molecular mechanisms of activated T cell death in vivo. Curr Opin Immunol 14(3):354–359

    Article  CAS  PubMed  Google Scholar 

  12. Trauth BC et al (1989) Monoclonal antibody-mediated tumor regression by induction of apoptosis. Science 245(4915):301–305

    Article  CAS  PubMed  Google Scholar 

  13. Krueger A, Fas SC, Baumann S, Krammer PH (2003) The role of CD95 in the regulation of peripheral T-cell apoptosis. Immunol Rev 193:58–69

    Article  CAS  PubMed  Google Scholar 

  14. Mateo V et al (2007) Perforin-dependent apoptosis functionally compensates Fas deficiency in activation-induced cell death of human T lymphocytes. Blood 110(13):4285–4292

    Article  CAS  PubMed  Google Scholar 

  15. Lanzarotti N et al (2014) RAS-associated lymphoproliferative disease evolves into severe juvenile myelo-monocytic leukemia. Blood 123(12):1960–1963

    Article  CAS  PubMed  Google Scholar 

  16. Niemela JE et al (2011) Somatic KRAS mutations associated with a human nonmalignant syndrome of autoimmunity and abnormal leukocyte homeostasis. Blood 117(10):2883–2886

    Article  PubMed  PubMed Central  Google Scholar 

  17. Oliveira JB et al (2007) NRAS mutation causes a human autoimmune lymphoproliferative syndrome. Proc Natl Acad Sci U S A 104(21):8953–8958

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Jodo S et al (2001) Apoptosis-inducing membrane vesicles. A novel agent with unique properties. J Biol Chem 276(43):39938–39944

    Article  CAS  PubMed  Google Scholar 

  19. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139(2):271–279

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. INSERM UMR 1163, Laboratory of The Immunogenetics of Pediatric Autoimmune Diseases, Paris, France

    Aude Magerus-Chatinet & Frédéric Rieux-Laucat

  2. Imagine Institute, Paris Descartes—Sorbonne Paris Cité University, 24 Boulevard de Montparnasse, 75015, Paris, France

    Aude Magerus-Chatinet & Frédéric Rieux-Laucat

Authors
  1. Aude Magerus-Chatinet
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Frédéric Rieux-Laucat
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Aude Magerus-Chatinet .

Editor information

Editors and Affiliations

  1. ER440-OSS, Label INSERM, Universite de Rennes CLCC Eugene Marquis, RENNES, France

    Patrick Legembre

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Magerus-Chatinet, A., Rieux-Laucat, F. (2017). In Vitro Evaluation of the Apoptosis Function in Human Activated T Cells. In: Legembre, P. (eds) CD95. Methods in Molecular Biology, vol 1557. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6780-3_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6780-3_4

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6778-0

  • Online ISBN: 978-1-4939-6780-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation