Granulocyte–colony-stimulatory factor: a strong inhibitor of natural killer cell function
Laura Schlahsa
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
Search for more papers by this authorYarúa Jaimes
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
Search for more papers by this authorRainer Blasczyk
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
Search for more papers by this authorConstança Figueiredo
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
Search for more papers by this authorLaura Schlahsa
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
Search for more papers by this authorYarúa Jaimes
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
Search for more papers by this authorRainer Blasczyk
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
Search for more papers by this authorConstança Figueiredo
From the Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
Search for more papers by this authorAbstract
BACKGROUND: The human cytokine granulocyte–colony stimulatory factor (G-CSF) has found widespread application in the medical treatment of neutropenia and to mobilize hematopoietic stem cells used for transplantation. So far, the effect of G-CSF on natural killer (NK) cells has not been fully investigated.
STUDY DESIGN AND METHODS: The effect of G-CSF on the phenotype, cytokine secretion profile, and cytotoxicity of NK cells was assessed. NK cells incubated in vitro in presence of G-CSF for 48 hours as well as NK cells isolated from peripheral blood of G-CSF–mobilized stem cell donors (in vivo) were used.
RESULTS: In vitro, G-CSF caused a strongly altered phenotype in NK cells with 49% down regulation of NKp44 frequency. Furthermore, the expression of the activating receptors NKp46 and NKG2D decreased 40 and 64%, respectively. The expression of KIR2DL1 and KIR2DL2 decreased by 46% each. In cytotoxicity assays, the lytic capacity of G-CSF–exposed NK cells is reduced by up to 68% in vitro and up to 83% in vivo. Accordingly, granzyme B levels of in vivo G-CSF–exposed NK cells were reduced by up to 87% in comparison to nonstimulated NK cells. Cytokine production of in vitro and in vivo incubated NK cells was strongly decreased for interferon-γ, tumor necrosis factor-α, and granulocyte macrophage colony-stimulating factor as well as interleukin (IL)-6 and IL-8. Furthermore, we observed a reduction in proliferation and a positive feedback loop that increased the expression of the G-CSF receptor.
CONCLUSION: G-CSF was demonstrated to be a strong inhibitor of NK cells activity and may prevent their graft-versus-leukemia effect after transplantation.
Supporting Information
Figure S1. Effect of G-CSF and a Non-specific control on NK cell phenotype. NK cells freshly isolated from healthy donors were incubated for 48 hours, with the cytokines: IL-2 plus IL-15, G-CSF alone, IL-2 plus IL-15 in combination with G-CSF, Non-specific control (NS), and IL-2 plus IL-15 in combination with NS. The frequency of NKp44 and the expression of NKG2A was analysed by FACS.
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REFERENCES
- 1 Pan L, Bressler S, Cooke KR, Krenger W, Karandikar M, Ferrara JL. Long-term engraftment, graft-vs.-host disease, and immunologic reconstitution after experimental transplantation of allogeneic peripheral blood cells from G-CSF-treated donors. Biol Blood Marrow Transplant 1996; 2: 126-33.
- 2 Lane TA, Law P, Maruyama M, Young D, Burgess J, Mullen M, Mealiffe M, Terstappen LW, Hardwick A, Moubayed M, et al. Harvesting and enrichment of hematopoietic progenitor cells mobilized into the peripheral blood of normal donors by granulocyte-macrophage colony-stimulating factor (GM-CSF) or G-CSF: potential role in allogeneic marrow transplantation. Blood 1995; 85: 275-82.
- 3 Klumpp TR, Mangan KF, Goldberg SL, Pearlman ES, MacDonald JS. Granulocyte colony-stimulating factor accelerates neutrophil engraftment following peripheral-blood stem-cell transplantation: a prospective, randomized trial. J Clin Oncol 1995; 13: 1323-7.
- 4 Ringden O, Labopin M, Gorin NC, Le Blanc K, Rocha V, Gluckman E, Reiffers J, Arcese W, Vossen JM, Jouet JP, Cordonnier C, Frassoni F. Treatment with granulocyte colony-stimulating factor after allogeneic bone marrow transplantation for acute leukemia increases the risk of graft-versus-host disease and death: a study from the Acute Leukemia Working Party of the European Group for Blood and Marrow Transplantation. J Clin Oncol 2004; 22: 416-23.
- 5 Ringden OT, Le Blanc K, Remberger M. Granulocyte and granulocyte-macrophage colony-stimulating factors in allografts: uses, misuses, misconceptions, and future applications. Exp Hematol 2005; 33: 505-12.
- 6 Gazitt Y. Immunologic profiles of effector cells and peripheral blood stem cells mobilized with different hematopoietic growth factors. Stem Cells 2000; 18: 390-8.
- 7 Rutella S, Zavala F, Danese S, Kared H, Leone G. Granulocyte colony-stimulating factor: a novel mediator of T cell tolerance. J Immunol 2005; 175: 7085-91.
- 8 Morris ES, MacDonald KP, Kuns RD, Morris HM, Banovic T, Don AL, Rowe V, Wilson YA, Raffelt NC, Engwerda CR, Burman AC, Markey KA, Godfrey DI, Smyth MJ, Hill GR. Induction of natural killer T cell-dependent alloreactivity by administration of granulocyte colony-stimulating factor after bone marrow transplantation. Nat Med 2009; 15: 436-41.
- 9 Tanaka J, Kobayashi S, Mori A, Ohta S, Yamamoto Y, Hashino S, Asaka M, Imamura M. Immunophenotype of peripheral blood mononuclear cells and NK cell activity after allogeneic bone marrow transplantation using recombinant human granulocyte colony-stimulating factor. Acta Haematol 1999; 102: 196-8.
- 10 Pavletic ZS, Joshi SS, Pirruccello SJ, Tarantolo SR, Kollath J, Reed EC, Bierman PJ, Vose JM, Warkentin PI, Gross TG, Nasrati K, Armitage JO, Kessinger A, Bishop MR. Lymphocyte reconstitution after allogeneic blood stem cell transplantation for hematologic malignancies. Bone Marrow Transplant 1998; 21: 33-41.
- 11 Ruggeri L, Capanni M, Urbani E, Perruccio K, Shlomchik WD, Tosti A, Posati S, Rogaia D, Frassoni F, Aversa F, Martelli MF, Velardi A. Effectiveness of donor natural killer cell alloreactivity in mismatched hematopoietic transplants. Science 2002; 295: 2097-100.
- 12 Ringden O, Karlsson H, Olsson R, Omazic B, Uhlin M. The allogeneic graft-versus-cancer effect. Br J Haematol 2009; 147: 614-33.
- 13 Pende D, Marcenaro S, Falco M, Martini S, Bernardo ME, Montagna D, Romeo E, Cognet C, Martinetti M, Maccario R, Mingari MC, Vivier E, Moretta L, Locatelli F, Moretta A. 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: 3119-29.
- 14 Yu J, Venstrom JM, Liu XR, Pring J, Hasan RS, O'Reilly RJ, Hsu KC. Breaking tolerance to self, circulating natural killer cells expressing inhibitory KIR for non-self HLA exhibit effector function after T cell-depleted allogeneic hematopoietic cell transplantation. Blood 2009; 113: 3875-84.
- 15 Hsu KC, Keever-Taylor CA, Wilton A, Pinto C, Heller G, Arkun K, O'Reilly RJ, Horowitz MM, Dupont B. Improved outcome in HLA-identical sibling hematopoietic stem-cell transplantation for acute myelogenous leukemia predicted by KIR and HLA genotypes. Blood 2005; 105: 4878-84.
- 16 Bignon JD, Gagne K. KIR matching in hematopoietic stem cell transplantation. Curr Opin Immunol 2005; 17: 553-9.
- 17 Farag SS, Fehniger T, Ruggeri L, Velardi A, Caligiuri MA. Natural killer cells: biology and application in stem-cell transplantation. Cytotherapy 2002; 4: 445-6.
- 18 Ruggeri L, Mancusi A, Perruccio K, Burchielli E, Martelli MF, Velardi A. Natural killer cell alloreactivity for leukemia therapy. J Immunother 2005; 28: 175-82.
- 19 Jedema I, van der Werff NM, Barge RM, Willemze R, Falkenburg JH. New CFSE-based assay to determine susceptibility to lysis by cytotoxic T cells of leukemic precursor cells within a heterogeneous target cell population. Blood 2004; 103: 2677-82.
- 20 Vago L, Forno B, Sormani MP, Crocchiolo R, Zino E, Di Terlizzi S, Lupo Stanghellini MT, Mazzi B, Perna SK, Bondanza A, Middleton D, Palini A, Bernardi M, Bacchetta R, Peccatori J, Rossini S, Roncarolo MG, Bordignon C, Bonini C, Ciceri F, Fleischhauer K. Temporal, quantitative, and functional characteristics of single-KIR-positive alloreactive natural killer cell recovery account for impaired graft-versus-leukemia activity after haploidentical hematopoietic stem cell transplantation. Blood 2008; 112: 3488-99.
- 21 Freedman MH, Bonilla MA, Fier C, Bolyard AA, Scarlata D, Boxer LA, Brown S, Cham B, Kannourakis G, Kinsey SE, Mori PG, Cottle T, Welte K, Dale DC. Myelodysplasia syndrome and acute myeloid leukemia in patients with congenital neutropenia receiving G-CSF therapy. Blood 2000; 96: 429-36.
- 22 Rosenberg PS, Alter BP, Bolyard AA, Bonilla MA, Boxer LA, Cham B, Fier C, Freedman M, Kannourakis G, Kinsey S, Schwinzer B, Zeidler C, Welte K, Dale DC; Severe Chronic Neutropenia International Registry. The incidence of leukemia and mortality from sepsis in patients with severe congenital neutropenia receiving long-term G-CSF therapy. Blood 2006; 107: 4628-35.
- 23 Beekman R, Touw IP. G-CSF and its receptor in myeloid malignancy. Blood 2010; 115: 531-6.
- 24 Freedman MH, Alter BP. Malignant myeloid transformation in congenital forms of neutropenia. Isr Med Assoc J 2002; 4: 1011-4.
- 25 Kudo K, Nagai H, Numata S, Ichihara M, Kinoshita T, Horibe K, Kato K, Matsuyama T, Kodera Y, Kojima S. Absence of mutations in the granulocyte colony-stimulating factor (G-CSF) receptor gene in patients with myelodysplastic syndrome/acute myeloblastic leukaemia occurring after treatment of aplastic anaemia with G-CSF. Br J Haematol 2000; 111: 656-8.
- 26 Bohlius J, Reiser M, Schwarzer G, Engert A. Granulopoiesis-stimulating factors to prevent adverse effects in the treatment of malignant lymphoma. Cochrane Database Syst Rev 2004; CD003189.
- 27 Ruggeri L, Capanni M, Casucci M, Volpi I, Tosti A, Perruccio K, Urbani E, Negrin RS, Martelli MF, Velardi A. Role of natural killer cell alloreactivity in HLA-mismatched hematopoietic stem cell transplantation. Blood 1999; 94: 333-9.
- 28 Miller JS, Soignier Y, Panoskaltsis-Mortari A, McNearney SA, Yun GH, Fautsch SK, McKenna D, Le C, Defor TE, Burns LJ, Orchard PJ, Blazar BR, Wagner JE, Slungaard A, Weisdorf DJ, Okazaki IJ, McGlave PB. Successful adoptive transfer and in vivo expansion of human haploidentical NK cells in patients with cancer. Blood 2005; 105: 3051-7.
- 29 Passweg JR, Stern M, Koehl U, Uharek L, Tichelli A. Use of natural killer cells in hematopoetic stem cell transplantation. Bone Marrow Transplant 2005; 35: 637-43.
- 30 Koehl U, Sorensen J, Esser R, Zimmermann S, Grüttner HP, Tonn T, Seidl C, Seifried E, Klingebiel T, Schwabe D. IL-2 activated NK cell immunotherapy of three children after haploidentical stem cell transplantation. Blood Cells Mol Dis 2004; 33: 261-6.
- 31 Velardi A, Ruggeri L, Mancusi A, Aversa F, Christiansen FT. Natural killer cell allorecognition of missing self in allogeneic hematopoietic transplantation: a tool for immunotherapy of leukemia. Curr Opin Immunol 2009; 21: 525-30.
- 32 Pende D, Rivera P, Marcenaro S, Chang CC, Biassoni R, Conte R, Kubin M, Cosman D, Ferrone S, Moretta L, Moretta A. Major histocompatibility complex class I-related chain A and UL16-binding protein expression on tumor cell lines of different histotypes: analysis of tumor susceptibility to NKG2D-dependent natural killer cell cytotoxicity. Cancer Res 2002; 62: 6178-86.
- 33 Lopez-Botet M, Bellon T. Natural killer cell activation and inhibition by receptors for MHC class I. Curr Opin Immunol 1999; 11: 301-7.
- 34 Byrd A, Hoffmann SC, Jarahian M, Momburg F, Watzl C. Expression analysis of the ligands for the Natural Killer cell receptors NKp30 and NKp44. Plos One 2007; 2: e1339.
- 35 Sloand EM, Kim S, Maciejewski JP, Van Rhee F, Chaudhuri A, Barrett J, Young NS. Pharmacologic doses of granulocyte colony-stimulating factor affect cytokine production by lymphocytes in vitro and in vivo. Blood 2000; 95: 2269-74.
- 36 Kitabayashi A, Hirokawa M, Hatano Y, Lee M, Kuroki J, Niitsu H, Miura AB. Granulocyte colony-stimulating factor downregulates allogeneic immune responses by posttranscriptional inhibition of tumor necrosis factor-alpha production. Blood 1995; 86: 2220-7.
- 37 Yoshimura T, Matsushima K, Tanaka S, Robinson EA, Appella E, Oppenheim JJ, Leonard EJ. Purification of a human monocyte-derived neutrophil chemotactic factor that has peptide sequence similarity to other host defense cytokines. Proc Natl Acad Sci U S A 1987; 84: 9233-7.
- 38 Walz A, Peveri P, Aschauer H, Baggiolini M. Purification and amino acid sequencing of NAF, a novel neutrophil-activating factor produced by monocytes. Biochem Biophys Res Commun 1987; 149: 755-61.
- 39 Nicola NA, Metcalf D. Binding of 125I-labeled granulocyte colony-stimulating factor to normal murine hemopoietic cells. J Cell Physiol 1985; 124: 313-21.
- 40 Lieschke GJ, Grail D, Hodgson G, Metcalf D, Stanley E, Cheers C, Fowler KJ, Basu S, Zhan YF, Dunn AR. Mice lacking granulocyte colony-stimulating factor have chronic neutropenia, granulocyte and macrophage progenitor cell deficiency, and impaired neutrophil mobilization. Blood 1994; 84: 1737-46.
- 41 Henslee PJ, Thompson JS, Romond EH, Doukas MA, Metcalfe M, Marshall ME, MacDonald JS. T cell depletion of HLA and haploidentical marrow reduces graft-versus-host disease but it may impair a graft-versus-leukemia effect. Transplant Proc 1987; 19(1 Pt 3): 2701-6.
- 42 Barfield RC, Otto M, Houston J, Holladay M, Geiger T, Martin J, Leimig T, Gordon P, Chen X, Handgretinger R. A one-step large-scale method for T- and B-cell depletion of mobilized PBSC for allogeneic transplantation. Cytotherapy 2004; 6: 1-6.