Polyfunctionality of natural killer cell in healthy donors

Yupanun WUTTI-IN; Preeyanat VONGCHAN; Hathairat THANANCHAI

Authors

Yupanun WUTTI-IN Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200
Preeyanat VONGCHAN Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200
Hathairat THANANCHAI Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200

Keywords:

Polyfunction, NK cells, healthy donor, innate immunity

Abstract

Background: Natural killer (NK) cells are important guards of the innate immune system, which act by performing as primary effector cells in viral infections. NK cell function is regulated by the engagement of activating and/or inhibitory receptors on individual NK cell surfaces. Subsequent to activation, the release of preformed cytolytic granules or cytokines occurs. Recently, the polyfunctionality of NK cells has been described as a potent NK cell subset that mediates antiviral response in HIV-infected slow progressors. Objectives: To evaluate the polyfunctional NK cells in healthy individuals. Methods: Peripheral blood mononuclear cells (PBMCs) were separated from 41 healthy blood donors by Ficoll-Hypaque gradient centrifugation. Multicolor flow cytometry was used to investigate the expression of function markers (degranulation marker (CD107a), IFN-γ, and TNF-a) on NK cells following PMA/Ionomycin or K562 stimulation. Results: The percentage of NK cells expressing CD107a, IFN-γ, or TNF-a in response to PMA/Ionomycin were 17.85, 10.56, and 2.66 %, respectively. The NK cells expressing CD107a, IFN-γ, or TNF-a in response to K562 stimulation were 6.43, 2.09, and 0.57 %, respectively. The capability of NK cells to perform polyfunctions was 6.19 % of the total NK cells following PMA/Ionomycin stimulation, while 1.06 % was observed following K562 stimulation. The trifunctional CD107a⁺ / IFN-γ⁺/ TNF-a ⁺ NK cell subset was found to be 0.95 and 0.04 % following PMA/Ionomycin and K562 stimulation, respectively. Conclusion: A small fraction of NK cells was capable of performing polyfunctions following stimulation, with less than 1 % being able to perform trifunctions in this study setting.

Downloads

Download data is not yet available.

Metrics

Metrics Loading ...

Author Biography

Yupanun WUTTI-IN, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200

Department of Medical Technology

References

MA Cooper, TA Fehniger and MA Caligiuri. The biology of human natural killer-cell subsets. Trends Immunol. 2001; 22, 633-40.

LL Lanier. NK cell recognition. Annu. Rev. Immunol. 2005; 23, 225-74.

MA Cooper, TA Fehniger, A Fuchs, M Colonna and MA Caligiuri. NK cell and DC interactions. Trends Immunol. 2004; 25, 47-52.

R Vankayalapati, A Garg, A Porgador, DE Griffith, P Klucar, H Safi, WM Girard, D Cosman, T Spies and PF Barnes. Role of NK cell-activating receptors and their ligands in the lysis of mononuclear phagocytes infected with an intracellular bacterium. J. Immunol. 2005; 175, 4611-7.

L Golden-Mason, L Madrigal-Estebas, E McGrath, MJ Conroy, EJ Ryan, JE Hegarty, C O'Farrelly and DG Doherty. Altered natural killer cell subset distributions in resolved and persistent hepatitis C virus infection following single source exposure. Gut 2008; 57, 1121-28.

JM Milush, S Lopez-Verges, VA York, SG Deeks, JN Martin, FM Hecht, LL Lanier and DF Nixon. CD56negCD16(+) NK cells are activated mature NK cells with impaired effector function during HIV-1 infection. Retrovirology 2013; 10, 158.

D Mavilio, G Lombardo, J Benjamin, D Kim, D Follman, E Marcenaro, MA O'Shea, A Kinter, C Kovacs, A Moretta and AS Fauci. Characterization of CD56-/CD16+ natural killer (NK) cells: a highly dysfunctional NK subset expanded in HIV-infected viremic individuals. Proc. Natl. Acad. Sci. USA 2005; 102, 2886-91.

Y Levy, R Thiebaut, M Montes, C Lacabaratz, L Sloan, B King, S Perusat, C Harrod, A Cobb, LK Roberts, M Surenaud, C Boucherie, S Zurawski, C Delaugerre, L Richert, G Chene, J Banchereau and K Palucka. Dendritic cell-based therapeutic vaccine elicits polyfunctional HIV-specific T-cell immunity associated with control of viral load. Eur. J. Immunol. 2014; 44, 2802-10.

J Lee, J Kim, SJ Shin and EC Shin. DNA immunization of Mycobacterium tuberculosis resuscitation-promoting factor B elicits polyfunctional CD8(+) T cell responses. Clin. Exp. Vaccine Res. 2014; 3, 235-43.

T Prezzemolo, G Guggino, MP La Manna, DD Liberto, F Dieli and N Caccamo. Functional Signatures of Human CD4 and CD8 T Cell Responses to Mycobacterium tuberculosis. Front. Immunol. 2014; 5, 180.

P Kamya, S Boulet, CM Tsoukas, JP Routy, R Thomas, P Cote, MR Boulassel, JG Baril, C Kovacs, SA Migueles, M Connors, TJ Suscovich, C Brander, CL Tremblay and N Bernard. Receptor-ligand requirements for increased NK cell polyfunctional potential in slow progressors infected with HIV-1 coexpressing KIR3DL1*h/*y and HLA-B*57. J. Virol. 2011; 85, 5949-60.

MS de Souza, C Karnasuta, AE Brown, LE Markowitz, S Nitayaphan, RP Garner, JG McNeil, DL Birx and JH Cox. A comparative study of the impact of HIV infection on natural killer cell number and function in Thais and North Americans. AIDS Res. Hum. Retroviruses 2000; 16, 1061-6.

AW Kay, J Fukuyama, N Aziz, CL Dekker, S Mackey, GE Swan, MM Davis, S Holmes and CA Blish. Enhanced natural killer-cell and T-cell responses to influenza A virus during pregnancy. Proc. Natl. Acad. Sci. USA 2014; 111, 14506-11.

H Horton, EP Thomas, JA Stucky, I Frank, Z Moodie, Y Huang, YL Chiu, MJ McElrath and SC De Rosa. Optimization and validation of an 8-color intracellular cytokine staining (ICS) assay to quantify antigen-specific T cells induced by vaccination. J. Immunol. Methods 2007; 323, 39-54.

HT Maecker, A Rinfret, P D'Souza, J Darden, E Roig, C Landry, P Hayes, J Birungi, O Anzala, M Garcia, A Harari, I Frank, R Baydo, M Baker, J Holbrook, J Ottinger, L Lamoreaux, CL Epling, E Sinclair, MA Suni, K Punt, S Calarota, S El-Bahi, G Alter, H Maila, E Kuta, J Cox, C Gray, M Altfeld, N Nougarede, J Boyer, L Tussey, T Tobery, B Bredt, M Roederer, R Koup, VC Maino, K Weinhold, G Pantaleo, J Gilmour, H Horton and RP Sekaly. Standardization of cytokine flow cytometry assays. BMC Immunol. 2005; 6, 13.

V Pascal, N Schleinitz, C Brunet, S Ravet, E Bonnet, X Lafarge, M Touinssi, D Reviron, JF Viallard, JF Moreau, J Dechanet-Merville, P Blanco, JR Harle, J Sampol, E Vivier, F Dignat-George and P Paul. Comparative analysis of NK cell subset distribution in normal and lymphoproliferative disease of granular lymphocyte conditions. Eur. J. Immunol. 2004; 34, 2930-40.

SS Farag and MA Caligiuri. Human natural killer cell development and biology. Blood Rev. 2006; 20, 123-37.

M Mitchai, N Leeratanapetch, V Lulitanond, P Srikoon, S Hattori, K Vaeteewoottacharn, S WongKham and S Okada. Phenotypic characteristic and function of NK cell subsets in cART-treated HIV-1 infected individuals. World J. AIDS 2014; 4, 293-300.

MA Caligiuri. Human natural killer cells. Blood. 2008; 112, 461-69.

N Zucchini, K Crozat, T Baranek, SH Robbins, M Altfeld and M Dalod. Natural killer cells in immunodefense against infective agents. Expert Rev. Anti Infect Ther. 2008; 6, 867-85.

G Alter, JM Malenfant and M Altfeld. CD107a as a functional marker for the identification of natural killer cell activity. J. Immunol. Methods 2004; 294, 15-22.