Functional characterization of NK cells in Mexican pediatric patients with acute lymphoblastic leukemia: Report from the Mexican Interinstitutional Group for the Identification of the Causes of Childhood Leukemia

Autoři: Lucero Valenzuela-Vazquez aff001;  Juan Carlos Núñez-Enríquez aff002;  Jacqueline Sánchez-Herrera aff001;  Elva Jiménez-Hernández aff003;  Jorge Alfonso Martín-Trejo aff004;  Laura Eugenia Espinoza-Hernández aff003;  Aurora Medina-Sanson aff005;  Luz Victoria Flores-Villegas aff006;  José Gabriel Peñaloza-González aff007;  José Refugio Torres-Nava aff008;  Rosa Martha Espinosa-Elizondo aff009;  Raquel Amador-Sánchez aff010;  Jessica Denisse Santillán-Juárez aff011;  Janet Flores-Lujano aff002;  María Luisa Pérez-Saldívar aff002;  Luis Ramiro García-López aff012;  Alejandro Castañeda-Echevarría aff013;  Francisco Rodríguez-Leyva aff014;  Haydeé Rosas-Vargas aff015;  Minerva Mata-Rocha aff015;  David Aldebarán Duarte-Rodríguez aff002;  Omar Alejandro Sepúlveda-Robles aff015;  Ismael Mancilla-Herrera aff016;  Juan Manuel Mejía-Aranguré aff017;  Mario Ernesto Cruz-Munoz aff001
Působiště autorů: Facultad de Medicina, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico aff001;  Unidad de Investigación Médica en Epidemiología Clínica, UMAE Hospital de Pediatría, Centro Médico Nacional (CMN) "Siglo XXI", Instituto Mexicano del Seguro Social (IMSS), Mexico City, Mexico aff002;  Servicio de Hematología Pediátrica, Hospital General “Gaudencio González Garza”, Centro Médico Nacional (CMN) "La Raza", IMSS, Mexico City, Mexico aff003;  Servicio de Hematología Pediátrica, UMAE Hospital de Pediatría, Centro Médico Nacional (CMN) "Siglo XXI", Instituto Mexicano del Seguro Social (IMSS), Mexico City, Mexico aff004;  Servicio de Hemato-Oncologia, Hospital Infantil de México Federico Gómez, Secretaria de Salud (SS), Mexico City, Mexico aff005;  Servicio de Hematología Pediátrica, Centro Médico Nacional (CMN) “20 de Noviembre”, Instituto de Seguridad Social al Servicio de los Trabajadores del Estado (ISSSTE), Mexico City, Mexico aff006;  Servicio de Onco-Pediatria, Hospital Juárez de México, Secretaria de Salud (SS), Mexico City, Mexico aff007;  Servicio de Oncología, Hospital Pediátrico de Moctezuma, Secretaría de Salud del D.F., Mexico City, Mexico aff008;  Servicio de Hematología Pediátrica, Hospital General de México, Secretaria de Salud (SS), Mexico City, Mexico aff009;  Hospital General Regional No. 1 "Carlos McGregor Sánchez Navarro", IMSS, Mexico City, Mexico aff010;  Servicio de Hemato-oncología Pediátrica, Hospital Regional No. 1° de Octubre, ISSSTE, Mexico City, Mexico aff011;  Servicio de Pediatría, Hospital Pediátrico de Tacubaya, Secretaría de Salud (SS), Mexico City, Mexico aff012;  Servicio de Pediatría, HGR No. 25, IMSS, Mexico City, Mexico aff013;  Servicio de Cirugía Pediátrica, HGZ No. 30 IMSS, Mexico City, Mexico aff014;  Unidad de Investigación Médica en Genética Humana, UMAE Hospital de Pediatría, Centro Médico Nacional (CMN) "Siglo XXI", IMSS, Mexico City, Mexico aff015;  Departamento de infectología e inmunología, Instituto Nacional de Perinatología, Mexico City, Mexico aff016;  Coordinación de Investigación en Salud, Instituto Mexicano del Seguro Social (IMSS), Mexico City, Mexico aff017
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: 10.1371/journal.pone.0227314


Acute lymphoblastic leukemia (ALL) is the most common cancer in children around the globe. Mexico City has one of the highest incidence rates of childhood leukemia worldwide with 49.5 cases per million children under the age of 15 which is similar to that reported for Hispanic populations living in the United States. In addition, it has been noted a dismal prognosis in Mexican and Hispanic ALL pediatric population. Although ALL, like cancer in general, has its origins in endogenous, exogenous, and genetic factors, several studies have shown that the immune system also plays a deterministic role in cancer development. Among various elements of the immune system, T lymphocytes and NK cells seem to dominate the immune response against leukemia. The aim of the present study was to perform a phenotypic and functional characterization of NK cells in ALL Mexican children at the moment of diagnosis and before treatment initiation. A case-control study was conducted by the Mexican Interinstitutional Group for the Identification of the Causes of Childhood Leukemia (MIGICCL). 41 cases were incident ALL children younger than 17 years old and residents of Mexico City. 14 controls were children without leukemia, matched by age and sex with cases. NK cell function was evaluated by degranulation assays towards K562 cells and SLAM-associated protein (SAP) expression was measured by intracellular staining. All assays were performed using peripheral blood mononuclear cells from controls and patients. The results indicate that NK mediated cytotoxicity, measured by CD107a degranulation assays in response to K562 cells, was reduced in ALL patients compared to controls. Interestingly, an impaired NK cell killing of target cells was not equally distributed among ALL patients. In contrast to patients classified as high-risk, standard-risk patients did not display a significant reduction in NK cell-mediated cytotoxicity. Moreover, patients presenting a leukocyte count ≥ 50,000xmm3 displayed a reduction in NK-cell mediated cytotoxicity and a reduction in SAP expression, indicating a positive correlation between a reduced SAP expression and an impaired NK cell-mediated citotoxicity. In the present study it was observed that unlike patients with standard-risk, NK cells from children presenting high-risk ALL, harbor an impaired cytotoxicity towards K562 at diagnosis. In addition, NK cell function was observed to be compromised in patients with a leukocyte count ≥50,000xmm3, where also it was noticed a decreased expression of SAP compared to patients with a leukocyte count <50,000xmm3. These data indicate NK cell-mediated cytotoxicity is not equally affected in ALL patients, nevertheless a positive correlation between low SAP expression and decreased NK cell-mediated cytotoxicity was observed in ALL patients with a leukocyte count ≥50,000xmm3. Finally, an abnormal NK cell-mediated cytotoxicity may represent a prognostic factor for high-risk acute lymphoblastic leukemia.

Klíčová slova:

Acute lymphoblastic leukemia – Cancer detection and diagnosis – Cell degranulation – Cytotoxicity – Leukemias – NK cells – Pediatrics – White blood cells


1. Greaves M. A causal mechanism for childhood acute lymphoblastic leukaemia. Nat Rev Cancer. 2018;18(8):471–84. Epub 2018/05/23. doi: 10.1038/s41568-018-0015-6 29784935.

2. Friedmann AM, Weinstein HJ. The role of prognostic features in the treatment of childhood acute lymphoblastic leukemia. Oncologist. 2000;5(4):321–8. Epub 2000/08/31. doi: 10.1634/theoncologist.5-4-321 10965000.

3. Carroll WL, Bhojwani D, Min DJ, Raetz E, Relling M, Davies S, et al. Pediatric acute lymphoblastic leukemia. Hematology Am Soc Hematol Educ Program. 2003:102–31. Epub 2003/11/25. doi: 10.1182/asheducation-2003.1.102 14633779.

4. Perez-Saldivar ML, Fajardo-Gutierrez A, Bernaldez-Rios R, Martinez-Avalos A, Medina-Sanson A, Espinosa-Hernandez L, et al. Childhood acute leukemias are frequent in Mexico City: descriptive epidemiology. BMC Cancer. 2011;11:355. Epub 2011/08/19. doi: 10.1186/1471-2407-11-355 21846410

5. Wilkinson JD, Gonzalez A, Wohler-Torres B, Fleming LE, MacKinnon J, Trapido E, et al. Cancer incidence among Hispanic children in the United States. Rev Panam Salud Publica. 2005;18(1):5–13. Epub 2005/08/18. doi: 10.1590/s1020-49892005000600002 16105320.

6. Abrahao R, Lichtensztajn DY, Ribeiro RC, Marina NM, Keogh RH, Marcos-Gragera R, et al. Racial/ethnic and socioeconomic disparities in survival among children with acute lymphoblastic leukemia in California, 1988–2011: A population-based observational study. Pediatr Blood Cancer. 2015;62(10):1819–25. Epub 2015/04/22. doi: 10.1002/pbc.25544 25894846.

7. Nunez-Enriquez JC, Barcenas-Lopez DA, Hidalgo-Miranda A, Jimenez-Hernandez E, Bekker-Mendez VC, Flores-Lujano J, et al. Gene Expression Profiling of Acute Lymphoblastic Leukemia in Children with Very Early Relapse. Arch Med Res. 2016;47(8):644–55. Epub 2017/05/10. doi: 10.1016/j.arcmed.2016.12.005 28476192.

8. Inaba H, Greaves M, Mullighan CG. Acute lymphoblastic leukaemia. Lancet. 2013;381(9881):1943–55. Epub 2013/03/26. doi: 10.1016/S0140-6736(12)62187-4 23523389

9. Palucka AK, Coussens LM. The Basis of Oncoimmunology. Cell. 2016;164(6):1233–47. Epub 2016/03/12. doi: 10.1016/j.cell.2016.01.049 26967289

10. Chiossone L, Dumas PY, Vienne M, Vivier E. Natural killer cells and other innate lymphoid cells in cancer. Nat Rev Immunol. 2018;18(11):671–88. Epub 2018/09/14. doi: 10.1038/s41577-018-0061-z 30209347.

11. Locatelli F, Pende D, Mingari MC, Bertaina A, Falco M, Moretta A, et al. Cellular and molecular basis of haploidentical hematopoietic stem cell transplantation in the successful treatment of high-risk leukemias: role of alloreactive NK cells. Front Immunol. 2013;4:15. Epub 2013/02/05. doi: 10.3389/fimmu.2013.00015 23378843

12. Locatelli F, Pende D, Maccario R, Mingari MC, Moretta A, Moretta L. Haploidentical hemopoietic stem cell transplantation for the treatment of high-risk leukemias: how NK cells make the difference. Clin Immunol. 2009;133(2):171–8. Epub 2009/06/02. doi: 10.1016/j.clim.2009.04.009 19481979.

13. Pende D, Marcenaro S, Falco M, Martini S, Bernardo ME, Montagna D, et al. Anti-leukemia activity of alloreactive NK cells in KIR ligand-mismatched haploidentical HSCT for pediatric patients: evaluation of the functional role of activating KIR and redefinition of inhibitory KIR specificity. Blood. 2009;113(13):3119–29. Epub 2008/10/24. doi: 10.1182/blood-2008-06-164103 18945967.

14. Maki G, Hayes GM, Naji A, Tyler T, Carosella ED, Rouas-Freiss N, et al. NK resistance of tumor cells from multiple myeloma and chronic lymphocytic leukemia patients: implication of HLA-G. Leukemia. 2008;22(5):998–1006. Epub 2008/02/22. doi: 10.1038/leu.2008.15 18288133.

15. Fauriat C, Just-Landi S, Mallet F, Arnoulet C, Sainty D, Olive D, et al. Deficient expression of NCR in NK cells from acute myeloid leukemia: Evolution during leukemia treatment and impact of leukemia cells in NCRdull phenotype induction. Blood. 2007;109(1):323–30. Epub 2006/08/31. doi: 10.1182/blood-2005-08-027979 16940427.

16. Sanchez-Correa B, Morgado S, Gayoso I, Bergua JM, Casado JG, Arcos MJ, et al. Human NK cells in acute myeloid leukaemia patients: analysis of NK cell-activating receptors and their ligands. Cancer Immunol Immunother. 2011;60(8):1195–205. Epub 2011/06/07. doi: 10.1007/s00262-011-1050-2 21644031.

17. Chang MC, Cheng HI, Hsu K, Hsu YN, Kao CW, Chang YF, et al. NKG2A Down-Regulation by Dasatinib Enhances Natural Killer Cytotoxicity and Accelerates Effective Treatment Responses in Patients With Chronic Myeloid Leukemia. Front Immunol. 2018;9:3152. Epub 2019/02/02. doi: 10.3389/fimmu.2018.03152 30705677

18. Tsirogianni M, Grigoriou E, Kapsimalli V, Dagla K, Stamouli M, Gkirkas K, et al. Natural killer cell cytotoxicity is a predictor of outcome for patients with high risk myelodysplastic syndrome and oligoblastic acute myeloid leukemia treated with azacytidine. Leuk Lymphoma. 2019:1–7. Epub 2019/04/06. doi: 10.1080/10428194.2019.1581935 30947589.

19. Diermayr S, Himmelreich H, Durovic B, Mathys-Schneeberger A, Siegler U, Langenkamp U, et al. NKG2D ligand expression in AML increases in response to HDAC inhibitor valproic acid and contributes to allorecognition by NK-cell lines with single KIR-HLA class I specificities. Blood. 2008;111(3):1428–36. Epub 2007/11/13. doi: 10.1182/blood-2007-07-101311 17993609.

20. Rouce RH, Shaim H, Sekine T, Weber G, Ballard B, Ku S, et al. The TGF-beta/SMAD pathway is an important mechanism for NK cell immune evasion in childhood B-acute lymphoblastic leukemia. Leukemia. 2016;30(4):800–11. Epub 2015/12/02. doi: 10.1038/leu.2015.327 26621337

21. Ramirez-Ramirez D, Padilla-Castaneda S, Galan-Enriquez CS, Vadillo E, Prieto-Chavez JL, Jimenez-Hernandez E, et al. CRTAM(+) NK cells endowed with suppressor properties arise in leukemic bone marrow. J Leukoc Biol. 2019;105(5):999–1013. Epub 2019/02/23. doi: 10.1002/JLB.MA0618-231R 30791148.

22. Moretta L, Pietra G, Montaldo E, Vacca P, Pende D, Falco M, et al. Human NK cells: from surface receptors to the therapy of leukemias and solid tumors. Front Immunol. 2014;5:87. Epub 2014/03/19. doi: 10.3389/fimmu.2014.00087 24639677

23. Long EO, Kim HS, Liu D, Peterson ME, Rajagopalan S. Controlling natural killer cell responses: integration of signals for activation and inhibition. Annu Rev Immunol. 2013;31:227–58. Epub 2013/03/23. doi: 10.1146/annurev-immunol-020711-075005 23516982

24. de Smith AJ, Walsh KM, Ladner MB, Zhang S, Xiao C, Cohen F, et al. The role of KIR genes and their cognate HLA class I ligands in childhood acute lymphoblastic leukemia. Blood. 2014;123(16):2497–503. Epub 2014/02/13. doi: 10.1182/blood-2013-11-540625 24518758

25. Lanier LL. NK cell recognition. Annu Rev Immunol. 2005;23:225–74. Epub 2005/03/18. doi: 10.1146/annurev.immunol.23.021704.115526 15771571.

26. Dong Z, Cruz-Munoz ME, Zhong MC, Chen R, Latour S, Veillette A. Essential function for SAP family adaptors in the surveillance of hematopoietic cells by natural killer cells. Nat Immunol. 2009;10(9):973–80. Epub 2009/08/04. doi: 10.1038/ni.1763 19648922.

27. Dong Z, Veillette A. How do SAP family deficiencies compromise immunity? Trends Immunol. 2010;31(8):295–302. Epub 2010/07/24. doi: 10.1016/ 20650688.

28. Veillette A. NK cell regulation by SLAM family receptors and SAP-related adapters. Immunol Rev. 2006;214:22–34. Epub 2006/11/15. doi: 10.1111/j.1600-065X.2006.00453.x 17100873.

29. Wu N, Zhong MC, Roncagalli R, Perez-Quintero LA, Guo H, Zhang Z, et al. A hematopoietic cell-driven mechanism involving SLAMF6 receptor, SAP adaptors and SHP-1 phosphatase regulates NK cell education. Nat Immunol. 2016;17(4):387–96. Epub 2016/02/16. doi: 10.1038/ni.3369 26878112.

30. Latour S, Veillette A. The SAP family of adaptors in immune regulation. Semin Immunol. 2004;16(6):409–19. Epub 2004/11/16. doi: 10.1016/j.smim.2004.08.020 15541655.

31. Francis SS, Wallace AD, Wendt GA, Li L, Liu F, Riley LW, et al. In utero cytomegalovirus infection and development of childhood acute lymphoblastic leukemia. Blood. 2017;129(12):1680–4. Epub 2016/12/17. doi: 10.1182/blood-2016-07-723148 27979823

32. Marcenaro S, Gallo F, Martini S, Santoro A, Griffiths GM, Arico M, et al. Analysis of natural killer-cell function in familial hemophagocytic lymphohistiocytosis (FHL): defective CD107a surface expression heralds Munc13-4 defect and discriminates between genetic subtypes of the disease. Blood. 2006;108(7):2316–23. Epub 2006/06/17. doi: 10.1182/blood-2006-04-015693 16778144.

33. Bryceson YT, Fauriat C, Nunes JM, Wood SM, Bjorkstrom NK, Long EO, et al. Functional analysis of human NK cells by flow cytometry. Methods Mol Biol. 2010;612:335–52. Epub 2009/12/25. doi: 10.1007/978-1-60761-362-6_23 20033652

34. Bryceson YT, Pende D, Maul-Pavicic A, Gilmour KC, Ufheil H, Vraetz T, et al. A prospective evaluation of degranulation assays in the rapid diagnosis of familial hemophagocytic syndromes. Blood. 2012;119(12):2754–63. Epub 2012/02/02. doi: 10.1182/blood-2011-08-374199 22294731.

35. Aggarwal N, Swerdlow SH, TenEyck SP, Boyiadzis M, Felgar RE. Natural killer cell (NK) subsets and NK-like T-cell populations in acute myeloid leukemias and myelodysplastic syndromes. Cytometry B Clin Cytom. 2016;90(4):349–57. Epub 2015/12/10. doi: 10.1002/cyto.b.21349 26648320.

36. Parrado A, Casares S, Rodriguez-Fernandez JM. Natural killer cytotoxicity and lymphocyte subpopulations in patients with acute leukemia. Leuk Res. 1994;18(3):191–7. Epub 1994/03/01. doi: 10.1016/0145-2126(94)90114-7 7511191.

37. Finn OJ. Immuno-oncology: understanding the function and dysfunction of the immune system in cancer. Ann Oncol. 2012;23 Suppl 8:viii6–9. Epub 2012/08/29. doi: 10.1093/annonc/mds256 22918931

38. Baier C, Fino A, Sanchez C, Farnault L, Rihet P, Kahn-Perles B, et al. Natural killer cells modulation in hematological malignancies. Front Immunol. 2013;4:459. Epub 2014/01/07. doi: 10.3389/fimmu.2013.00459 24391641

39. Coffey AJ, Brooksbank RA, Brandau O, Oohashi T, Howell GR, Bye JM, et al. Host response to EBV infection in X-linked lymphoproliferative disease results from mutations in an SH2-domain encoding gene. Nat Genet. 1998;20(2):129–35. Epub 1998/10/15. doi: 10.1038/2424 9771704.

40. Sayos J, Wu C, Morra M, Wang N, Zhang X, Allen D, et al. The X-linked lymphoproliferative-disease gene product SAP regulates signals induced through the co-receptor SLAM. Nature. 1998;395(6701):462–9. Epub 1998/10/17. doi: 10.1038/26683 9774102.

41. Smith MA, Seibel NL, Altekruse SF, Ries LA, Melbert DL, O’Leary M, et al. Outcomes for children and adolescents with cancer: challenges for the twenty-first century. J Clin Oncol. 2010;28(15):2625–34. Epub 2010/04/21. doi: 10.1200/JCO.2009.27.0421 20404250

42. Linabery AM, Ross JA. Trends in childhood cancer incidence in the U.S. (1992–2004). Cancer. 2008;112(2):416–32. Epub 2007/12/13. doi: 10.1002/cncr.23169 18074355.

43. Cruz-Munoz ME, Valenzuela-Vazquez L, Sanchez-Herrera J, Santa-Olalla Tapia J. From the "missing self" hypothesis to adaptive NK cells: Insights of NK cell-mediated effector functions in immune surveillance. J Leukoc Biol. 2019;105(5):955–71. Epub 2019/03/09. doi: 10.1002/JLB.MR0618-224RR 30848847.

44. Stary J, Zimmermann M, Campbell M, Castillo L, Dibar E, Donska S, et al. Intensive chemotherapy for childhood acute lymphoblastic leukemia: results of the randomized intercontinental trial ALL IC-BFM 2002. J Clin Oncol. 2014;32(3):174–84. Epub 2013/12/18. doi: 10.1200/JCO.2013.48.6522 24344215.

45. Mizia-Malarz A, Sobol-Milejska G. NK Cells as Possible Prognostic Factor in Childhood Acute Lymphoblastic Leukemia. Dis Markers. 2019;2019:3596983. Epub 2019/02/06. doi: 10.1155/2019/3596983 30719179

46. Bekker-Mendez VC, Miranda-Peralta E, Nunez-Enriquez JC, Olarte-Carrillo I, Guerra-Castillo FX, Pompa-Mera EN, et al. Prevalence of gene rearrangements in Mexican children with acute lymphoblastic leukemia: a population study-report from the Mexican Interinstitutional Group for the identification of the causes of childhood leukemia. Biomed Res Int. 2014;2014:210560. Epub 2015/02/19. doi: 10.1155/2014/210560 25692130

47. Schuster V, Kreth HW. X-linked lymphoproliferative disease is caused by deficiency of a novel SH2 domain-containing signal transduction adaptor protein. Immunol Rev. 2000;178:21–8. Epub 2001/02/24. doi: 10.1034/j.1600-065x.2000.17819.x 11213803.

48. Pachlopnik Schmid J, Canioni D, Moshous D, Touzot F, Mahlaoui N, Hauck F, et al. Clinical similarities and differences of patients with X-linked lymphoproliferative syndrome type 1 (XLP-1/SAP deficiency) versus type 2 (XLP-2/XIAP deficiency). Blood. 2011;117(5):1522–9. Epub 2010/12/02. doi: 10.1182/blood-2010-07-298372 21119115.

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