Reduced expansion of CD94/NKG2C+ NK cells in chronic lymphocytic leukemia and CLL-like monoclonal B-cell lymphocytosis is not related to increased human cytomegalovirus seronegativity or NKG2C deletions
Anna Puiggros
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorGonzalo Blanco
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorAura Muntasell
Hospital del Mar Medical Research Institute (IMIM), Barcelona, Spain
Search for more papers by this authorMaría Rodríguez-Rivera
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorMireia Altadill
University Pompeu Fabra (UPF), Barcelona, Spain
Search for more papers by this authorEulàlia Puigdecanet
MARGenomics, IMIM, Barcelona, Spain
Faculty of Medicine, University of Vic-Central University of Catalonia (UVic-UCC), Barcelona, Spain
Search for more papers by this authorXavier Calvo
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorEva Gimeno
Hematology Department, Hospital del Mar-IMIM, Barcelona, Spain
Applied Clinical Research in Hematological Malignances, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorEugènia Abella
Hematology Department, Hospital del Mar-IMIM, Barcelona, Spain
Applied Clinical Research in Hematological Malignances, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorPau Abrisqueta
Hematology Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain
Search for more papers by this authorFrancesc Bosch
Hematology Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain
Search for more papers by this authorJosé Yélamos
Immunology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Search for more papers by this authorAna Ferrer
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorMiguel López-Botet
Hospital del Mar Medical Research Institute (IMIM), Barcelona, Spain
University Pompeu Fabra (UPF), Barcelona, Spain
Search for more papers by this authorCorresponding Author
Blanca Espinet
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Correspondence
Blanca Espinet, Laboratori de Citogenètica Molecular, Servei de Patologia, Hospital del Mar, Pg. Marítim 25-29, 08003 Barcelona, Spain.
Email: [email protected]
Search for more papers by this authorAnna Puiggros
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorGonzalo Blanco
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorAura Muntasell
Hospital del Mar Medical Research Institute (IMIM), Barcelona, Spain
Search for more papers by this authorMaría Rodríguez-Rivera
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorMireia Altadill
University Pompeu Fabra (UPF), Barcelona, Spain
Search for more papers by this authorEulàlia Puigdecanet
MARGenomics, IMIM, Barcelona, Spain
Faculty of Medicine, University of Vic-Central University of Catalonia (UVic-UCC), Barcelona, Spain
Search for more papers by this authorXavier Calvo
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorEva Gimeno
Hematology Department, Hospital del Mar-IMIM, Barcelona, Spain
Applied Clinical Research in Hematological Malignances, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorEugènia Abella
Hematology Department, Hospital del Mar-IMIM, Barcelona, Spain
Applied Clinical Research in Hematological Malignances, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorPau Abrisqueta
Hematology Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain
Search for more papers by this authorFrancesc Bosch
Hematology Department, Hospital Universitari Vall d'Hebron, Barcelona, Spain
Search for more papers by this authorJosé Yélamos
Immunology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Search for more papers by this authorAna Ferrer
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Search for more papers by this authorMiguel López-Botet
Hospital del Mar Medical Research Institute (IMIM), Barcelona, Spain
University Pompeu Fabra (UPF), Barcelona, Spain
Search for more papers by this authorCorresponding Author
Blanca Espinet
Molecular Cytogenetics Laboratory, Hematological Cytology Laboratory, Pathology Department, Hospital del Mar, Barcelona, Spain
Translational Research on Hematological Neoplasms Group, Cancer Research Program, IMIM-Hospital del Mar, Barcelona, Spain
Correspondence
Blanca Espinet, Laboratori de Citogenètica Molecular, Servei de Patologia, Hospital del Mar, Pg. Marítim 25-29, 08003 Barcelona, Spain.
Email: [email protected]
Search for more papers by this authorAnna Puiggros and Gonzalo Blanco contributed equally to this study.
Abstract
Introduction
Dysregulated NK cell-mediated immune responses contribute to tumor evasion in chronic lymphocytic leukemia (CLL), although the NK cell compartment in CLL-like monoclonal B-cell lymphocytosis (MBL) is poorly understood. In healthy individuals, human cytomegalovirus (HCMV) induces the expansion of NK cells expressing high levels of CD94/NKG2C NK cell receptor (NKR) specific for HLA-E.
Methods
We analyzed the expression of NKG2A, NKG2C, ILT2, KIR, CD161, and CD57 in 24 MBL and 37 CLL. NKG2C was genotyped in these patients and in 81 additional MBL/CLL, while NKG2C gene expression was assessed in 26 cases. In 8 CLL patients with increased lymphocytosis (≥20 × 109/L), tumor HLA-E and HLA-G expression was evaluated.
Results
NKR distribution did not significantly differ between MBL and CLL patients, although they exhibited reduced NKG2C+ NK cells compared with a non-CLL group (4.6% vs 12.2%, P = .012). HCMV+ patients showed increased percentages of NKG2C+ NK cells compared with HCMV− (7.3% vs 2.9%, P = .176). Frequencies of NKG2C deletions in MBL/CLL were similar to those of the general population. Low/undetectable NKG2C expression was found among NKG2C+/− (45%) and NKG2C+/+ (12%) patients. CLL cases with increased lymphocytosis displayed especially reduced NKG2C expression (1.8% vs 8.1%, P = .029) and tumor cells with high HLA-E (>98%) and variable HLA-G expression (12.4%, range: 0.5-56.4). CLL patients with low NKG2C expression (<7%) showed shorter time to first treatment (P = .037).
Conclusion
Reduced percentages of CD94/NKG2C+ NK cells were observed in CLL and MBL patients independently of HCMV serostatus and NKG2C zygosity, particularly in CLL patients with increased lymphocytosis, which could potentially be related to the exposure to tumor cells.
CONFLICTS OF INTEREST
No potential conflicts of interest were disclosed.
Open Research
DATA AVAILABILITY STATEMENT
The data that support the findings of this study are available from the corresponding author upon reasonable request.
Supporting Information
| Filename | Description |
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| ijlh13494-sup-0001-Supinfo.docxapplication/word, 3.5 MB | Supplemental Methods |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- 1Ramsay AG, Clear AJ, Fatah R, Gribben JG. Multiple inhibitory ligands induce impaired T-cell immunologic synapse function in chronic lymphocytic leukemia that can be blocked with lenalidomide: establishing a reversible immune evasion mechanism in human cancer. Blood. 2012; 120(7): 1412-1421. https://doi.org/10.1182/blood-2012-02-411678
- 2Görgün G, Holderried TA, Zahrieh D, Neuberg D, Gribben JG. Chronic lymphocytic leukemia cells induce changes in gene expression of CD4 and CD8 T cells. J Clin Invest. 2005; 115(7): 1797-1805. https://doi.org/10.1172/JCI24176
- 3Riches JC, Davies JK, McClanahan F, et al. T cells from CLL patients exhibit features of T-cell exhaustion but retain capacity for cytokine production. Blood. 2013; 121(9): 1612-1621. https://doi.org/10.1182/blood-2012-09-457531
- 4Veuillen C, Aurran-Schleinitz T, Castellano R, et al. Primary B-CLL resistance to NK cell cytotoxicity can be overcome in vitro and in vivo by priming NK cells and monoclonal antibody therapy. J Clin Immunol. 2012; 32(3): 632-646. https://doi.org/10.1007/s10875-011-9624-5
- 5Huergo-Zapico L, Acebes-Huerta A, Gonzalez-Rodriguez AP, et al. Expansion of NK cells and reduction of NKG2D expression in chronic lymphocytic leukemia. Correlation with progressive disease. PLoS One. 2014; 9(10):e108326. https://doi.org/10.1371/journal.pone.0108326
- 6Parry HM, Stevens T, Oldreive C, et al. NK cell function is markedly impaired in patients with chronic lymphocytic leukaemia but is preserved in patients with small lymphocytic lymphoma. Oncotarget. 2016; 7(42): 68513-68526. https://doi.org/10.18632/oncotarget.12097
- 7MacFarlane AW, Jillab M, Smith MR, et al. NK cell dysfunction in chronic lymphocytic leukemia is associated with loss of the mature cells expressing inhibitory killer cell Ig-like receptors. Oncoimmunology. 2017; 6(7):e1330235. https://doi.org/10.1080/2162402X.2017.1330235
- 8Hofland T, Endstra S, Gomes CKP, et al. Natural Killer cell hypo-responsiveness in chronic lymphocytic leukemia can be circumvented in vitro by adequate activating signaling. Hemasphere. 2019; 3(6):e308. https://doi.org/10.1097/HS9.0000000000000308
- 9Rawstron AC, Bennett FL, O'Connor SJ, et al. Monoclonal B-cell lymphocytosis and chronic lymphocytic leukemia. N Engl J Med. 2008; 359(6): 575-583. https://doi.org/10.1056/NEJMoa075290
- 10Vardi A, Dagklis A, Scarfò L, et al. Immunogenetics shows that not all MBL are equal: the larger the clone, the more similar to CLL. Blood. 2013; 121(22): 4521-4528. https://doi.org/10.1182/blood-2012-12-471698
- 11Blanco G, Puiggros A, Sherry B, et al. Chronic lymphocytic leukemia-like monoclonal B-cell lymphocytosis exhibits an increased inflammatory signature that is reduced in early-stage chronic lymphocytic leukemia. Exp Hematol. 2021; 1: 1-13. https://doi.org/10.1016/j.exphem.2020.12.007
- 12D'Arena G, Rossi G, Minervini MM, et al. Circulating regulatory T cells in “clinical” monoclonal B-cell lymphocytosis. Int J Immunopathol Pharmacol. 2011; 24(4): 915-923. https://doi.org/10.1177/039463201102400410
- 13Rissiek A, Schulze C, Bacher U, et al. Multidimensional scaling analysis identifies pathological and prognostically relevant profiles of circulating T-cells in chronic lymphocytic leukemia. Int J Cancer. 2014; 135(10): 2370-2379. https://doi.org/10.1002/ijc.28884
- 14Blanco G, Vardi A, Puiggros A, et al. Restricted T cell receptor repertoire in CLL-like monoclonal B cell lymphocytosis and early stage CLL. Oncoimmunology. 2018; 7(6):e1432328. https://doi.org/10.1080/2162402X.2018.1432328
- 15te Raa GD, Tonino SH, Remmerswaal EB, et al. Chronic lymphocytic leukemia specific T-cell subset alterations are clone-size dependent and not present in monoclonal B lymphocytosis. LeukLymphoma. 2012; 53(11): 2321-2325. https://doi.org/10.3109/10428194.2012.698277
- 16Pourgheysari B, Bruton R, Parry H, et al. The number of cytomegalovirus-specific CD4+ T cells is markedly expanded in patients with B cell chronic lymphocytic leukemia and determines the total CD4+ T cell repertoire. Blood. 2010; 116(16): 2968-2974. https://doi.org/10.1182/blood-2009-12-257147
- 17te Raa GD, Pascutti MF, García-Vallejo JJ, et al. CMV-specific CD8+ T cell function is not impaired in chronic lymphocytic leukemia. Blood. 2014; 123(5): 717-724. https://doi.org/10.1182/blood-2013-08-518183
- 18Gumá M, Angulo A, Vilches C, et al. Imprint of human cytomegalovirus infection on the NK cell receptor repertoire. Blood. 2004; 104(12): 3664-3671. https://doi.org/10.1182/blood-2004-05-2058
- 19López-Botet M, Muntasell A, Vilches C. The CD94/NKG2C+ NK cell subset on the edge of innate and adaptive immunity to human cytomegalovirus infection. Semin Immunol. 2014; 26(2): 145-151. https://doi.org/10.1016/j.smim.2014.03.002
- 20Braud VM, Allan DS, O'Callaghan CA, et al. HLA-E binds to natural killer cell receptors CD94/NKG2A, B and C. Nature. 1998; 391(6669): 795-799. https://doi.org/10.1038/35869
- 21Tomasec P, Braud VM, Rickards C, et al. Surface expression of HLA-E, an inhibitor of natural killer cells, enhanced by human cytomegalovirus gpUL40. Science. 2000; 287(5455): 1031. https://doi.org/10.1126/science.287.5455.1031
- 22Hammer Q, Rückert T, Borst EM, et al. Peptide-specific recognition of human cytomegalovirus strains controls adaptive natural killer cells. Nat Immunol. 2018; 19(5): 453-463. https://doi.org/10.1038/s41590-018-0082-6
- 23Muntasell A, López-Montañés M, Vera A, et al. NKG2C zygosity influences CD94/NKG2C receptor function and the NK-cell compartment redistribution in response to human cytomegalovirus. Eur J Immunol. 2013; 43(12): 3268-3278. https://doi.org/10.1002/eji.201343773
- 24Muntasell A, Vilches C, Angulo A, López-Botet M. Adaptive reconfiguration of the human NK-cell compartment in response to cytomegalovirus: a different perspective of the host-pathogen interaction. Eur J Immunol. 2013; 43(5): 1133-1141. https://doi.org/10.1002/eji.201243117
- 25Goodier MR, White MJ, Darboe A, et al. Rapid NK cell differentiation in a population with near-universal human cytomegalovirus infection is attenuated by NKG2C deletions. Blood. 2014; 124(14): 2213-2222. https://doi.org/10.1182/blood-2014-05-576124
- 26Ataya M, Redondo-Pachón D, Llinàs-Mallol L, et al. Pretransplant adaptive NKG2C+ NK cells protect against cytomegalovirus infection in kidney transplant recipients. Am J Transplant. 2020; 20(3): 663-676. https://doi.org/10.1111/ajt.15658
- 27Petersen L, Roug AS, Skovbo A, Thysen AH, Eskelund CW, Hokland ME. TheCD94/NKG2C-expressing NK cell subset is augmented in chronic lymphocytic leukemia patients with positive human cytomegalovirus serostatus. Viral Immunol. 2009; 22(5): 333-337. https://doi.org/10.1089/vim.2009.0032
- 28Sanchez-Correa B, Morgado S, Gayoso I, et al. Human NK cells in acute myeloidleukaemia patients: analysis of NK cell-activating receptors and their ligands. Cancer Immunol Immunother. 2011; 60(8): 1195-1205. https://doi.org/10.1007/s00262-011-1050-2
- 29Moraru M, Cañizares M, Muntasell A, de Pablo R, López-Botet M, Vilches C. Assessment of copy-number variation in the NKG2C receptor gene in a single-tube and characterization of a reference cell panel, using standard polymerase chain reaction. Tissue Antigens. 2012; 80(2): 184-187. https://doi.org/10.1111/j.1399-0039.2012.01911.x
- 30Erikci AA, Karagoz B, Ozyurt M, Ozturk A, Kilic S, Bilgi O. HLA-G expression in B chronic lymphocytic leukemia: a new prognostic marker? Hematology. 2009; 14(2): 101-105. https://doi.org/10.1179/102453309X385197
- 31Costello RT, Knoblauch B, Sanchez C, Mercier D, Le Treut T, Sébahoun G. Expression of natural killer cell activating receptors in patients with chronic lymphocytic leukaemia. Immunology. 2012; 135(2): 151-157. https://doi.org/10.1111/j.1365-2567.2011.03521.x
- 32Muntasell A, Pupuleku A, Cisneros E, et al. Relationship of NKG2C copy number with the distribution of distinct cytomegalovirus-induced adaptive NK cell subsets. J Immunol. 2016; 196(9): 3818-3827. https://doi.org/10.4049/jimmunol.1502438
- 33Campos C, Pera A, Sanchez-Correa B, et al. Effect of age and CMV on NK cell subpopulations. Exp Gerontol. 2014; 54: 130-137. https://doi.org/10.1016/j.exger.2014.01.008
- 34Nückel H, Rebmann V, Dürig J, Dührsen U, Grosse-Wilde H. HLA-G expression is associated with an unfavorable outcome and immunodeficiency in chronic lymphocytic leukemia. Blood. 2005; 105(4): 1694-1698. https://doi.org/10.1182/blood-2004-08-3335
- 35Rezvany MR, Kazemi A, Hajifathali A, Kaviani S, Mellstedt H. Analysis of HLA-G gene expression in B-lymphocytes from chronic lymphocytic leukemia patients. Iran Biomed J. 2007; 11(2): 125-129.
- 36Giannopoulos K, Dmoszyńska A, Bojarska-Junak A, Schmitt M, Roliński J. Expression of HLA-G in patients with B-cell chronic lymphocytic leukemia (B-CLL). Folia Histochem Cytobiol. 2008; 46(4): 457-460. https://doi.org/10.2478/v10042-008-0072-x
- 37Rizzo R, Audrito V, Vacca P, et al. HLA-G is a component of the chronic lymphocytic leukemia escape repertoire to generate immune suppression: impact of the HLA-G 14 base pair (rs66554220) polymorphism. Haematologica. 2014; 99(5): 888-896. https://doi.org/10.3324/haematol.2013.095281
- 38Llano M, Lee N, Navarro F, et al. HLA-E-bound peptides influence recognition by inhibitory and triggering CD94/NKG2 receptors: preferential response to an HLA-G-derived nonamer. Eur J Immunol. 1998; 28(9): 2854-2863. https://doi.org/10.1002/(SICI)1521-4141(199809)28:09<2854:AID-IMMU2854>3.0.CO;2-W
10.1002/(SICI)1521-4141(199809)28:09<2854::AID-IMMU2854>3.0.CO;2-W CAS PubMed Web of Science® Google Scholar
- 39Valés-Gómez M, Reyburn HT, Erskine RA, López-Botet M, Strominger JL. Kinetics and peptide dependency of the binding of the inhibitory NK receptor CD94/NKG2-A and the activating receptor CD94/NKG2-C to HLA-E. EMBO J. 1999; 18(15): 4250-4260. https://doi.org/10.1093/emboj/18.15.4250
- 40Lauterbach N, Wieten L, Popeijus HE, Voorter CE, Tilanus MG. HLA-E regulates NKG2C+ natural killer cell function through presentation of a restricted peptide repertoire. Hum Immunol. 2015; 76(8): 578-586. https://doi.org/10.1016/j.humimm.2015.09.003
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