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Assessment of Natural Killer Cell Responses to Human Cytomegalovirus-Infected Macrophages

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Virus-Host Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1064))

Abstract

Natural killer (NK) cells are major components of the innate immune system and are assumed to play an important role in the defense against cytomegalovirus infection. Human cytomegalovirus (HCMV) is the only virus known that shapes the NK cell receptor repertoire in humans. Assays allowing the simultaneous measurement of multiple cell parameters and the assessment of subpopulations of NK cells are suitable for monitoring the NK cell response to infected cells. Herein, we describe an autologous assay system to assess NK cell responses to HCMV-infected macrophages which is based on flow cytometry to detect CD107a expression and interferon gamma (IFNγ) secretion. Further we established a simple method to handle HCMV-infected macrophages. Our assay provides a practicable approach to investigate NK cell responses to HCMV infection in vitro.

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References

  1. Biron CA, Byron KS, Sullivan JL (1989) Severe herpesvirus infections in an adolescent without natural killer cells. N Engl J Med 320(26):1731–1735

    Article  CAS  PubMed  Google Scholar 

  2. Orange JS (2006) Human natural killer cell deficiencies. Curr Opin Allergy Clin Immunol 6(6):399–409

    Article  PubMed  Google Scholar 

  3. Quinnan GV Jr, Kirmani N, Rook AH et al (1982) Cytotoxic T cells in cytomegalovirus infection: HLA-restricted T-lymphocyte and non-T-lymphocyte cytotoxic responses correlate with recovery from cytomegalovirus infection in bone-marrow-transplant recipients. N Engl J Med 307(1):7–13

    Article  PubMed  Google Scholar 

  4. Kuijpers TW, Baars PA, Dantin C et al (2008) Human NK cells can control CMV infection in the absence of T cells. Blood 112(3):914–915

    Article  CAS  PubMed  Google Scholar 

  5. Wilkinson GW, Tomasec P, Stanton RJ et al (2008) Modulation of natural killer cells by human cytomegalovirus. J Clin Virol 41(3):206–212

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Orange JS, Fassett MS, Koopman LA et al (2002) Viral evasion of natural killer cells. Nat Immunol 3(11):1006–1012

    Article  CAS  PubMed  Google Scholar 

  7. Romo N, Magri G, Muntasell A et al (2011) Natural killer cell-mediated response to human cytomegalovirus-infected macrophages is modulated by their functional polarization. J Leukoc Biol 90(4):717–726

    Article  CAS  PubMed  Google Scholar 

  8. Sinclair J, Sissons P (2006) Latency and reactivation of human cytomegalovirus. J Gen Virol 87(Pt 7):1763–1779

    Article  CAS  PubMed  Google Scholar 

  9. Sinzger C, Hahn G, Digel M et al (2008) Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E. J Gen Virol 89(Pt 2):359–368

    Article  CAS  PubMed  Google Scholar 

  10. Alter G, Malenfant JM, Altfeld M (2004) CD107a as a functional marker for the identification of natural killer cell activity. J Immunol Methods 294(1–2):15–22

    Article  CAS  PubMed  Google Scholar 

  11. Tomescu C, Chehimi J, Maino VC et al (2009) Retention of viability, cytotoxicity, and response to IL-2, IL-15, or IFN-alpha by human NK cells after CD107a degranulation. J Leukoc Biol 85(5):871–876

    Article  CAS  PubMed  Google Scholar 

  12. Al-Hubeshy ZB, Coleman A, Nelson M et al (2011) A rapid method for assessment of natural killer cell function after multiple receptor crosslinking. J Immunol Methods 366(1–2):52–59

    Article  CAS  PubMed  Google Scholar 

  13. Warren HS (2011) Target-induced natural killer cell loss as a measure of NK cell responses. J Immunol Methods 370(1–2):86–92

    Article  CAS  PubMed  Google Scholar 

  14. Duske H, Sputtek A, Binder T et al (2011) Assessment of physiologic natural killer cell cytotoxicity in vitro. Hum Immunol 72(11):1007–1012

    Article  CAS  PubMed  Google Scholar 

  15. Schuessler A, Laib Sampaio K, Sinzger C (2010) Improvement of a quantitative cell tropism assay for rapid and reliable characterization of human cytomegalovirus mutants. J Virol Methods 167(2):218–222

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

The authors thank Ingrid Bennett for secretarial work and colleagues at our institution for discussion and technical help.

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Wu, Z., Frascaroli, G., Mertens, T. (2013). Assessment of Natural Killer Cell Responses to Human Cytomegalovirus-Infected Macrophages. In: Bailer, S., Lieber, D. (eds) Virus-Host Interactions. Methods in Molecular Biology, vol 1064. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-601-6_21

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  • DOI: https://doi.org/10.1007/978-1-62703-601-6_21

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-600-9

  • Online ISBN: 978-1-62703-601-6

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