Skip to main content
SpringerLink
Log in

Psychoneuroimmunology and Natural Killer Cells: The Chromium Release Whole Blood Assay

  • Protocol
  • First Online:
Psychoneuroimmunology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 934))

Abstract

Natural killer (NK) cells are an essential component of innate immunity. These lymphocytes are also sensitive barometers of the effects of endogenous and exogenous stressors on the immune system. This chapter will describe a chromium (51Cr) release bioassay designed to measure the target cell killing capacity of NK cells (NKCC). Key features of the cytotoxicity assay are that it is done with whole blood and that numbers of effector cells are determined for each sample by flow cytometry and lymphocyte count. Effector cells are defined as CD3−CD56+ lymphocytes. Target cells are the K562 eyrthroleukemia cell line. Killing capacity is defined as number of target cells killed per effector cell, at an effector cell/target cell ratio of 1:1 during a 4 h in vitro assay.

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 159.00
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. Herberman RB, Holden HT (1979) Natural killer cells as antitumor effector cells. J Natl Cancer Inst 62:441–445

    PubMed  CAS  Google Scholar 

  2. Solomon GF, Benton D, Harker JO, Bonavida B, Fletcher MA (1994) Prolonged asymptomatic states in HIV-seropositive persons with fewer than 50 CD4+ T cells per MM3. Psychoneuroimmunologic findings. Ann N Y Acad Sci 741:85–90

    Article  Google Scholar 

  3. Su DM, Vankayalapati R (2010) A new avenue to cure cancer by turning adaptive immune T cells to innate immune NK cells via reprogramming. J Mol Cell Biol 2:237–239

    Article  PubMed  CAS  Google Scholar 

  4. Ironson G, Wyings C, Schneiderman N et al (1997) Post-traumatic stress symptoms, intrusive thoughts, loss and immune function after Hurricane Andrew. Psychosom Med 59:128–141

    PubMed  CAS  Google Scholar 

  5. Schedlowski M, Jacobs R, Stratmann G et al (1993) Changes of natural killer cells during acute psychological stress. J Clin Immunol 13:119–126

    Article  PubMed  CAS  Google Scholar 

  6. Segerstrom SC, Miller GE (2004) Psychological stress and the human immune system: a meta-analytic study of 30 years of inquiry. Psychol Bull 130:601–630

    Article  PubMed  Google Scholar 

  7. Klimas NG, Rosenthal M, Fletcher MA (2006) Immune effects of an acute exercise challenge in Gulf War Illness. Assoc Military Surgeons US, San Antonio

    Google Scholar 

  8. Harvey JM, Barnes Z, Zeng et al (2011) Exercise effects on biomarkers of CFS/ME and GWI. Presented to the International Conference on CFS/ME, Ottawa, Canada

    Google Scholar 

  9. Cohen S, Tyrrell DA, Smith AP (1991) Psychological stress and susceptibility to the common cold. N Engl J Med 325:606–612

    Article  PubMed  CAS  Google Scholar 

  10. Okamoto H, Tsunoda T, Teruya K et al (2008) An occupational health study of emergency physicians in Japan: health assessment by immune variables (CD4, CD8, CD56, and NK cell activity) at the beginning of work. J Occup Health 50:136–146

    Article  PubMed  Google Scholar 

  11. Fletcher MA, Zeng XR, Maher K et al (2010) Biomarkers in chronic fatigue syndrome: evaluation of natural killer cell function and dipeptidyl peptidase IV/CD26. PLoS One 5(5):e10817

    Article  PubMed  Google Scholar 

  12. Maher K, Klimas NG, Fletcher MA (2005) Chronic fatigue syndrome is associated with diminished intracellular perforin. Clin Exp Immunol 142:505–511

    PubMed  CAS  Google Scholar 

  13. Alter G, Malenfant JM, Altfeld M (2004) CD107a as a functional marker for the identification of natural killer cell activity. J Immunol Methods 294:15–22

    Article  PubMed  CAS  Google Scholar 

  14. Brunner K, Mauel TJ, Cerottini J-C et al (1968) Quantitative assay of the lytic action of immune lymphoid cells on Cr labeled allogeneic target cells in vitro Inhibition by isoantibody and by drugs. Immunology 14:181

    PubMed  CAS  Google Scholar 

  15. Baron GA, Klimas NG, Fischl MA et al (1985) Decreased natural killer cell-mediated cytotoxicity per effector cell. Diagn Immunol 3:197–204

    PubMed  CAS  Google Scholar 

  16. Fletcher MA, Baron GA, Ashman MA et al (1987) Use of whole blood methods in assessment of immune parameters in immunodeficiency syndromes. Diagn Clin Immunol 5:69–81

    PubMed  CAS  Google Scholar 

  17. Bourin P, Mansour I, Doinel C, Roué R et al (1993) Circadian rhythms of circulating NK cells in healthy and human immunodeficiency virus-infected men. Chronobiol Int 10:298–305

    Article  PubMed  CAS  Google Scholar 

  18. Cleland WW (1967) The statistical analysis of enzyme kinetic data. Adv Enzymol Relat Areas Mol Biol 29:1

    PubMed  CAS  Google Scholar 

  19. Callewaert DM, Mahle NH (1985) Kinetic models of natural killer cells and their use for studying activated NK cells. In: Herberman RB, Callewaert DM (eds) Mechanisms of cytotoxicity by NK cells. Academic, Orlando, p 381

    Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the NIAAA: R21AA016635 (PI MA Fletcher); NIAID: R01AI065723 (PI MA Fletcher); CFIDS Assoc. of America: (PI N Klimas); NIAID: UO1 AI459940 (PI N Klimas); NIAMSD R01 AR057853-01A1 (PI N Klimas); VA merit awards (PI N Klimas) Dynamic Modeling in GWI, CFS/ME; GW080152 (PI N Klimas).

Author information

Authors and Affiliations

  1. Department of Medicine, University of Miami, Miami, FL, USA

    Mary Ann Fletcher & Zachary Barnes

  2. Miami Veterans Affairs Medical Center, Miami, FL, USA

    Mary Ann Fletcher & Nancy G. Klimas

  3. Department of Medicine, Division of Pulmonary Medicine, University of Alberta, Edmonton, AB, Canada

    Gordon Broderick

  4. Institute for Neuro-immune Medicine, Nova Southeastern University, Ft. Lauderdale, FL, USA

    Nancy G. Klimas

Authors
  1. Mary Ann Fletcher
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zachary Barnes
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gordon Broderick
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nancy G. Klimas
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mary Ann Fletcher .

Editor information

Editors and Affiliations

  1. PharmTao, Lafayette St 4601, Santa Clara, 95056, California, USA

    Qing Yan

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Fletcher, M.A., Barnes, Z., Broderick, G., Klimas, N.G. (2012). Psychoneuroimmunology and Natural Killer Cells: The Chromium Release Whole Blood Assay. In: Yan, Q. (eds) Psychoneuroimmunology. Methods in Molecular Biology, vol 934. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-071-7_16

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-071-7_16

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-070-0

  • Online ISBN: 978-1-62703-071-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation