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Chapter 11.13 : Natural Killer Cell Assays

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Abstract:

Natural killer (NK) cells are another group of cytolytic lymphocytes, distinct from B lymphocytes and T lymphocytes, that participate in both innate immunity and adaptive immunity. NK cells are lymphocytes that lack B-cell receptors and T-cell receptors. They are designed to kill certain mutant cells and virus-infected cells. Normally present as a small subpopulation of circulating lymphocytes, NK cells morphologically appear as so-called “large granular lymphocytes.” Like cytotoxic T cells, they attack and kill tumor cells and protect against a wide variety of infectious microbes. They are “natural” killers because they do not need additional stimulation or to recognize a specific antigen in order to attack and kill. NK cells appear to play a role in a variety of human diseases. Compromised or absent natural immunity, as measured in vitro by decreased NK activity and/or depressed absolute numbers of circulating NK cells, has been linked to the development and progression of cancer, chronic and acute viral infections (including AIDS), various immunodeficiencies, and certain autoimmune diseases.

Citation: Garcia L. 2010. Natural Killer Cell Assays, p 219-231. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch11.13
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Figures

Image of Figure 11.13.2-1
Figure 11.13.2-1

NK cell panel interpretation. As per the 2005 Practice Parameter for the Diagnosis and Management of Primary Immunodeficiency, a mutation in the Fc gamma R3III gene affecting NK cell function can be screened for by demonstrating decreased detection of CD16 using the B73.1 clone. If the CD16-positive population is detectable using both the anti-CD16 clones (B73.1 and 3G8) it is unlikely that the individual is homozygous for this mutation.

Citation: Garcia L. 2010. Natural Killer Cell Assays, p 219-231. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch11.13
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Image of Figure 11.13.2-2
Figure 11.13.2-2

Example of a normal CD45/side scatter histogram.

Citation: Garcia L. 2010. Natural Killer Cell Assays, p 219-231. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch11.13
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Download as Powerpoint

References

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1. Comans-Bitter, W. M., et al. 1997. Immunophenotyping of blood lymphocytes in childhood. Reference values for lymphocyte subpopulations. J. Pediatr. 130:388393.
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3. Parks, D. R.,, L. A. Herzenberg,, and L. Herzenberg,. 1989. Flow cytometry and fluorescence-activated cell sorting, p. 781802. In W. E. Paul (ed.), Fundamental Immunology, 2nd ed. Raven Press, New York, NY.
4. Shapiro, H. M.,, E. R. Schildkraut,, R. Curbelo,, R. B. Turner, et al. 1977. Cytomat-R: a computer-controlled multiple laser source multiparameter flow cytophotometer system. J. Histochem. Cytochem. 25:836844.
1. Bryant, J.,, R. Day,, T. L. Whiteside,, and R. B. Herbeman. 1992. Calculation of lytic units for the expression of cell-mediated cytotoxicity. J. Immunol. Methods 146:91103.
2. Douglas, S. D.,, S. Durako,, N. B. Tustin,, J. Houser,, L. Muenz,, S. E. Starr,, and C. Wilson for the Adolescent Medicine HIV/AIDS Research Network. 2001. Natural killer cell enumeration and function in HIV-infected and high-risk uninfected adolescents. AIDS Res. Hum. Retrovir. 17:543552.
3. Evans, D.,, K. Lynch,, T. Benton,, B. Dube,, D. Gettes,, N. Tustin,, J. Lai,, D. Metzger,, and S. Douglas. 2008. Selective serotonin reuptake inhibitor and substance P antagonist enhancement of natural killer cell innate immunity in human immunodeficiency virus/acquired immunodeficienty syndrome. Biol. Psychiatr. 63:899905.
4. Hay, R.,, J. Caputo,, T. R. Chen,, M. Macy,, P. McClintock,, and Y. Reid (ed.). 1994. American Type Culture Collection (ATCC) Catalogue of Cell Lines and Hybridomas, 8th ed. 1994 reference guide. American Type Culture Collection, Manassas, VA.
5. Lozzio, C. B.,, and B. B. Lozzio. 1975. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood 45:321334.
6. Whiteside, T.,, C. R. Rindalo,, and R. B. Herberman,. 1992. Cytolytic cell functions, p. 220230. In N. R. Rose,, E. Conway de Macario,, J. L. Fahey,, H. Friedman,, and G. M. Penn (ed.), Manual of Clinical Laboratory Immunology, 4th ed. American Society for Microbiology, Washington, DC.
7. Whiteside, T. L., 2006. Measurement of NK cell activity in humans, p. 296300. In B. Detrick,, R. G. Hamilton,, and J. D. Folds (ed.), Manual of Clinical Laboratory Immunology, 7th ed. ASM Press, Washington, DC.
1.. Hay, R.,, J. Caputo,, T. R. Chen,, M. Macy,, P. McClintock,, and Y. Reid. 1994. American Type Culture Collection (ATCC) Catalogue of Cell Lines and Hybridomas, 8th ed. 1994 reference guide. American Type Culture Collection, Manassas, VA.
2.. Lozzio, C. B.,, and B. B. Lozzio. 1975. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood 45:321334.

Tables

Generic image for table
Table 11.13.2-1

Example of tube labeling for NK cell flow cytometry assay

MsIgG1, mouse immunoglobulin G1; FITC, fluorescein isothiocyanate; PE, phycoerythrin; ECD, phycoerythrin with Texas Red.

Citation: Garcia L. 2010. Natural Killer Cell Assays, p 219-231. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch11.13
Generic image for table
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Citation: Garcia L. 2010. Natural Killer Cell Assays, p 219-231. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch11.13

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