Preview

Medical Immunology (Russia)

Advanced search

HETEROGENEITY OF NK AND NKT LYMPHOCYTE POPULATIONS IN HEALTHY DONORS

https://doi.org/10.15789/1563-0625-2017-4-401-408

Abstract

Natural killer(NK) and  NKT lymphocytes are important components of innate immunity, and compose a first-line defense against cancer. These populations are characterized by high heterogeneity and are divided into several subpopulations, by differences in functional activity as well as CD56  and CD16  expression. Studying  heterogeneity for these  lymphocyte populations in healthy  donors  is important, due  to imbalance between  different  lymphocyte subsets in cancer patients. Changes in the ratio of these subpopulations may be of prognostic and clinical  significance in malignant diseases. The present  study was conducted with peripheral blood  lymphocytes in 50 healthy  donors. When  analysing  population of NK  lymphocytes we have identified 18.0±11.3% of antigen-positive cells, their fluctuations ranged  from 7.6 to 29.2%, whereas average number of cells with  CD3-CD56+ and  CD3-CD16+   phenotypes was equal  to 16,2±8.1%, and  11,0±6.7%, respectively. The  subpopulation analysis  showed  that  the  primary  pool  of NK  cells  was presented by CD56dimCD16dim cells  by  52.3±19.9 percent.  We  detected  minor   subpopulations,  e.g.,  CD56dimCD16bright,  CD56-CD16+, CD56brightCD16- (0.3±0.2%, 1.7±0.9%, and  1.3±0.6%,  respectively). Search  for  intracellular perforin has revealed  that the number of CD56+Perf+ cells comprized 25.1±14.8%, CD16+Perf+, 23.8±16.0%. Cytometric analysis showed that perforin is found, predominantly, in CD56dimCD16dim NK lymphocytes, whereas the cells with CD56dimCD16bright, CD56-CD16+, CD56brightCD16- immunophenotypes did not produce perforin. For the first time, we have discovered a subpopulation of NK cells with the СD56dimCD16dim immunophenotype that did not contain intracellular perforin (2.0%).  The NKT cell population with СD3+CD16/СD56+  phenotype was detected in 7.1% (25% – 3.45; 75% – 8.75) antigen-positive cells, within a range of 2.5 to 11.9%. Analysis with a combination of monoclonal antibodies CD3/CD56/CD16 has shown that the number of CD3+ CD56+ cells was 4.33% (25% – 2.25; 75% – 7.3), whereas the number of CD3+CD16+ was 3.087% (25% – 0.9; 75% – 6.2). These  data  demonstrate that  the differences in results  between  the CD3/CD16/CD56, and  CD3/CD16 test systems are statistically significant  (p < 0.05). It was shown that the primary-pool NKT-cells are CD56+CD16- cells, whose number is about 5.45% (25% – 2.95; 75% – 7.3) among  total CD3+ lymphocyte population. Two minor subpopulations were also detected which differed in expression  of CD56 and CD16 antigens. Hence, the level of CD56-CD16+ cells was 3% (25% – 0.25; 75% – 3.05),  and the number of CD56+CD16+ was equal to 0.67% (25% – 0.25; 75% – 0.9). Hence, the observed wide phenotypic diversity of NK and NKT-cells reflects their  ability  to exert  various  functional activities.  This  study, showing  high  heterogeneity of NK  and  NKT lymphocytes, may serve as a basis for the study of imbalances between  different  subpopulations of these cells in cancer patients.

About the Authors

D V. Tabakov
Research Institute of Clinical Oncology, N. Blokhin Research Center for Oncology
Russian Federation

Tabakov  Dmitry V. - Research Associate, Laboratory of Clinical Immunology of Tumors.

115478, Moscow, Kashirskoye ch., 23. Phone: 7 (962) 956-25-97



T. N. Zabotina
Research Institute of Clinical Oncology, N. Blokhin Research Center for Oncology
Russian Federation

PhD, MD (Biology), Leading Research Associate, Laboratory of Clinical Immunology of Tumors.

Moscow



A. A. Borunova
Research Institute of Clinical Oncology, N. Blokhin Research Center for Oncology
Russian Federation

PhD (Medicine), Senior Research Associate, Laboratory of Clinical Immunology of Tumors.

Moscow



I. O. Panchuk
Research Institute of Clinical Oncology, N. Blokhin Research Center for Oncology
Russian Federation

Resident Physician, Laboratory of Clinical Immunology of Tumors.

Moscow



O. V. Korotkova
Research Institute of Clinical Oncology, N. Blokhin Research Center for Oncology
Russian Federation

PhD (Medicine), Senior Research Associate, Laboratory of Clinical Immunology of Tumors.

Moscow



Z. G. Kadagidze
Research Institute of Clinical Oncology, N. Blokhin Research Center for Oncology
Russian Federation

PhD, MD (Medicine), Professor, Head, Centralized Clinical Laboratory Department, Laboratory of Clinical Immunology of Tumors.

Moscow



References

1. Балмасова И.П., Ющук Н.Д., Знойко О.О., Шмелева Е.В., Дунда Н.И., Еремина О.Ф., Максимов С.Л., Зайцева М.Н., Малова Е.С., Сепиашвили Я.Р. Соотношение субпопуляций ЕК/ЕКТ как прогностический критерий хронического гепатита С // Аллергология и иммунология, 2009. Т. 10, № 3. С. 340-343. [Balmasova I.P., Yuschuk N.D., Znoiko O.O., Shmelyova E.V., Dunda N.I., Eryomina O.F., Maksimov S.L., Zaytseva M.N., Malova E.S., Sepiashvili Ya.R. NK/NKT ratio as a prognostic marker of chronic hepatitis C. Allergologiya i immunologiya = Allergology and Immunology, 2009, Vol. 10, no. 3, pp. 340-343. (In Russ.)]

2. Борунова А.А., Чкадуа Г.З., Заботина Т.Н. Перфорин-опосредованная цитотоксичность СD16+лимфоцитов // Иммунология, 2006. Т. 27, № 1. С. 4-6. [Borunova A.A., Chkadua G.Z., Zabotina Т.N. Perforinmedialing cytotoxicity of CD16+-lymphocytes. Immunologiya = Immunology, 2006, Vol. 27, no.1, pp. 4-6. (In Russ.)]

3. Заботина Т.Н., Короткова О.В., Борунова А.А., Очеева Н.Ю., Бокин И.И., Жорданиа К.И., Паниченко И.В., Сельчук В.Ю., Кузнецов В.В., Кадагидзе З.Г. Субпопуляционная структура лимфоцитов у больных раком яичников // Вестник РОНЦ им. Н.Н. Блохина РАМН, 2010. Т. 21, № 1. С. 46-52. [Zabotina T.N., Korotkova O.V., Borunova A.A., Ocheyeva N.Yu., Bokin I.I., Zhordania K.I., Panichenko I.V., Selchuk V.Yu., Kuznetsov V.V., Kadagidze Z.G. Lymphocyte subset structure in patients with ovarian cancer. Vestnik RONTS im. N.N. Blokhina RAMN = Journal of N.N. Blokhin Russian Cancer Research Center, 2010, Vol. 21, no. 1, pp. 46-52. (In Russ.)]

4. Короткова О.В., Заботина Т.Н., Скотаренко Л.В., Борунова А.А., Очеева Н.Ю., Воротников И.К., Кадагидзе З.Г. Субпопуляции лимфоцитов периферической крови больных РМЖ // Российский биотерапевтический журнал, 2011. Т. 10, № 3. С. 95-98. [Korotkova O.V., Zabotina T.N., Skotarenko L.V., Borunova A.A., Ocheeva N.Yu., Vorotnikov I.K., Kadagidze Z.G. Lymphocytes subpopulation from breast cancer patient’s peripheral blood. Rossiyskiy bioterapevticheskiy zhurnal = Russian Biotherapeutic Journal, 2011, Vol. 10, no. 3, pp. 95-98. (In Russ.)]

5. Biron C.A., Nguyen K.B., Pien G.C., Cousens L.P., Salazar-Mather T.P. Natural killer cells in antiviral defense: function and regulation by innate cytokines. Annu. Rev. Immunol., 1999, Vol. 17, pp. 189-220.

6. Cooper M.A., Fehniger T.A., Caligiuri M.A. The biology of human natural killer-cell subsets. Trends Immunol., 2001, Vol. 22, no. 11, pp. 633-640.

7. Cristiani C.M., Palella E., Sottile R., Tallerico R., Garofalo C., Carbone E. Human NK Cell Subsets in Pregnancy and Disease: Toward a New Biological Complexity. Front. Immunol., 2016, Vol. 7, p. 656.

8. De Maria A., Bozzano F., Cantoni C., Moretta L. Revisiting human natural killer cell subset function revealed cytolytic CD56dimCD16+ NK cells as rapid producers of abundant IFN-γ on activation. Proceedings of the National Academy of Sciences of the United States of America, 2011, Vol. 108, no. 22, pp. 728-732.

9. Del Zotto G., Marcenaro E., Vacca P., Sivori S., Pende D., Della Chiesa M., Moretta F., Ingegnere T., Mingari M.C., Moretta A., Moretta L. Markers and function of human nk cells in normal and pathological conditions. Cytometry B Clin Cytom. 2017 Jan 5. doi: 10.1002/cyto.b.21508. [Epub ahead of print]

10. East J.E., Kennedy A.J., Webb T.J. Raising the Roof: The Preferential Pharmacological Stimulation of Th1 and Th2 Response Mediated by NKT Cells. Med. Res. Rev., 2014, Vol. 34, no. 1, pp. 45-76.

11. Fauriat C., Long E.O., Ljunggren H.-G., Bryceson Y.T. Regulation of human NK-cell cytokine and chemokine production by target cell recognition. Blood, 2010, Vol. 115, no. 11, pp. 2167-2176.

12. Freud A.G., Caligiuri M.A. Human natural killer cell development. Immunological Reviews, 2006, Vol. 214, no. 1, pp. 56-72.

13. Gabrielli S., Ortolani C., Del Zotto G., Luchetti F., Canonico B., Buccella F., Artico M., Papa S., Zamai L. The Memories of NK Cells: Innate-Adaptive Immune Intrinsic Crosstalk. J. Immunol. Res., 2016, Vol. 2016, 1376595. doi: 0.1155/2016/1376595. Epub 2016 Dec 19.

14. Gross C., Schulte-Mecklenbeck A., Wiendl H., Marcenaro E., Kerlero de Rosbo N., Uccelli A., Laroni A. Regulatory Functions of Natural Killer Cells in Multiple Sclerosis. Front. Immunol., 2016, Vol. 7, p. 606.

15. Hazenberg M.D., Spits H. Human innate lymphoid cells. Blood, 2014, Vol. 124, no. 5, pp. 700-709.

16. Jiang Y., Cui X., Cui C., Zhang J., Zhou F., Zhang Z., Fu Y., Xu J., Chu Z., Liu J., Han X., Liao C., Wang Y., Cao Y., Shang H.. The Function of CD3+CD56+ NKT-Like Cells in HIV-Infected Individuals. Biomed. Res. Int., 2014, 863625. Published online 2014 Mar 20.

17. Jiao Y., Huntington N.D., Belz G.T., Seillet C. Type 1 Innate Lymphoid Cell Biology: Lessons Learnt from Natural Killer Cells. Front. Immunol., 2016, Vol. 7, p. 426.

18. Lanier L.L. Shades of grey-the blurring view of innate and adaptive immunity. Nature Reviews Immunology, 2013, Vol. 13, no. 2, pp. 73-74.

19. Lanier L.L., Chang C., Azuma M., Ruitenberg J.J., Hemperly J.J., Phillips J.H. Molecular and functional analysis of human natural killer cell-associated neural cell adhesion molecule (N-CAM/CD56). J. Immunol., 1991, Vol. 146, no. 12, pp. 4421-4426.

20. Littwitz-Salomon E., Dittmer U., Sutter K. Insufficient natural killer cell responses against retroviruses: how to improve NK cell killing of retrovirus-infected cells. Retrovirology, 2016, Vol. 13, p. 77.

21. Lundgren S., Warfvinge C.F., Elebro J., Heby M., Nodin B., Krzyzanowska A., Bjartell A., Leandersson K., Eberhard J., Jirström K. The Prognostic Impact of NK/NKT Cell Density in Periampullary Adenocarcinoma Differs by Morphological Type and Adjuvant Treatment. PLoS One, 2016, Vol. 11, no. 6, e0156497. doi: 10.1371/journal.pone.0156497.

22. Mace E.M., Orange J.S. Genetic Causes of Human NK Cell Deficiency and Their Effect on NK Cell Subsets. Front. Immunol., 2016, Vol. 7, p. 545.

23. Moretta A., Bottino C., Vitale M., Pende D., Cantoni C., Mingari M.C. Activating receptors and coreceptors involved in human natural killer cell-mediated cytolysis. Annu. Rev. Immunol., 2001, Vol. 19, pp. 197-223.

24. Nowak M., Schmidt-Wolf I.G.H. Natural Killer T Cells Subsets in Cancer, Functional Defects in Prostate Cancer and Implications for Immunotherapy. Cancers (Basel), 2011, Vol. 3, no. 3, pp. 3661-3675.

25. Pesce S., Moretta L., Moretta A., and Marcenaro E. Human NK Cell Subsets Redistribution in Pathological Conditions: A Role for CCR7 Receptor. Front. Immunol., 2016, Vol. 7, p. 414.

26. Poli A., Michel T., Thérésine M., Andrès E., Hentges F., Zimmer J. CD56bright natural killer (NK) cells: an important NK cell subset. Immunology, 2009, Vol. 126, no. 4, pp. 458-465.

27. Shevtsov M., Multhoff G. Immunological and Translational Aspects of NK Cell-Based Antitumor Immunotherapies. Front. Immunol., 2016, Vol. 11, no. 7, p. 492.

28. Viale R., Ware R., Maricic I., Chaturvedi V., Kumar V. NKT Cell Subsets Can Exert Opposing Effects in Autoimmunity, Tumor Surveillance and Inflammation. Curr. Immunol. Rev., 2012, Vol. 8, no. 4, pp. 287-296.

29. Zamai L., Del Zotto G., Buccella F., Galeotti L., Canonico B., Luchetti F., Papa S. Cytotoxic functions and susceptibility to apoptosis of human CD56bright NK cells differentiated in vitro from CD34+ hematopoietic progenitors. Cytometry Part A, 2012, Vol. 81, no. 4, pp. 294-302.


Review

For citations:


Tabakov D.V., Zabotina T.N., Borunova A.A., Panchuk I.O., Korotkova O.V., Kadagidze Z.G. HETEROGENEITY OF NK AND NKT LYMPHOCYTE POPULATIONS IN HEALTHY DONORS. Medical Immunology (Russia). 2017;19(4):401-408. (In Russ.) https://doi.org/10.15789/1563-0625-2017-4-401-408

Views: 2115


Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.


ISSN 1563-0625 (Print)
ISSN 2313-741X (Online)