Skip to main content
Erschienen in: Journal of Cancer Research and Clinical Oncology 12/2016

10.09.2016 | Original Article – Clinical Oncology

Cryopreserved NK cells in the treatment of haematological malignancies: preclinical study

verfasst von: Monika Holubova, Michaela Miklikova, Martin Leba, Daniel Georgiev, Pavel Jindra, Martin Caprnda, Rachele Ciccocioppo, Peter Kruzliak, Daniel Lysak

Erschienen in: Journal of Cancer Research and Clinical Oncology | Ausgabe 12/2016

Einloggen, um Zugang zu erhalten

Abstract

Background

Leukaemia is an aggressive cancer of haematopoiesis. Despite increasing treatment success, the relapse rate is still high. Natural killer (NK) cells play a key role in the immune response to malignancies; thus, it is conceivable that NK cell-based immunotherapy may control relapses, while extending the disease-free survival. In our study, we investigated whether cryopreserved NK cells are able to kill the leukaemic K562 cell line, the necessity of IL-2 co-application and the association of activation marker expression (NKp44, NKG2D and CD25) with cytotoxic potential.

Materials and methods

K562 cells were added to NK cell cultures in different ratios, i.e. 1:5, 1:10 and 1:20 (K562/NK), immediately after thawing NK cells or after 3–6–12–24 h of re-cultivation with or without IL-2.

Results

Our results demonstrated the ability of cryopreserved NK cells to kill K562 in all ratios, times and culture conditions. The number of dead K562 cells depended on the number of NK cells and on the presence of IL-2. NK cells cytotoxic potential decreased gradually in the culture without IL-2. In contrast, NK cell-mediated cytotoxicity remained the same during the entire re-culture period after IL-2 re-application.

Conclusion

Our study proved the efficacy of using cryopreserved ready-for-use NK cells in relapse treatment and the need for simultaneous administration of IL-2.
Literatur
Zurück zum Zitat Bebenek M, Dus D, Kozlak J (2007) Fas/Fas-ligand expressions in primary breast cancer are significant predictors of its skeletal spread. Anticancer Res 27:215–218PubMed Bebenek M, Dus D, Kozlak J (2007) Fas/Fas-ligand expressions in primary breast cancer are significant predictors of its skeletal spread. Anticancer Res 27:215–218PubMed
Zurück zum Zitat Biassoni R, Bottino C, Cantoni C, Moretta A (2002) Human natural killer receptors and their ligands. Curr Protoc Immunol Chapter 14:Unit 14.10 Biassoni R, Bottino C, Cantoni C, Moretta A (2002) Human natural killer receptors and their ligands. Curr Protoc Immunol Chapter 14:Unit 14.10
Zurück zum Zitat Campbell KS, Purdy AK (2011) Structure/function of human killer cell immunoglobulin-like receptors: lessons from polymorphisms, evolution, crystal structures and mutations. Immunology 132:315–325CrossRefPubMedPubMedCentral Campbell KS, Purdy AK (2011) Structure/function of human killer cell immunoglobulin-like receptors: lessons from polymorphisms, evolution, crystal structures and mutations. Immunology 132:315–325CrossRefPubMedPubMedCentral
Zurück zum Zitat Cantoni C, Bottino C, Vitale M, Pessino A, Augugliaro R, Malaspina A, Parolini S, Moretta L, Moretta A, Biassoni R (1999) NKp44, a triggering receptor involved in tumor cell lysis by activated human natural killer cells, is a novel member of the immunoglobulin superfamily. J Exp Med 189:787–796CrossRefPubMedPubMedCentral Cantoni C, Bottino C, Vitale M, Pessino A, Augugliaro R, Malaspina A, Parolini S, Moretta L, Moretta A, Biassoni R (1999) NKp44, a triggering receptor involved in tumor cell lysis by activated human natural killer cells, is a novel member of the immunoglobulin superfamily. J Exp Med 189:787–796CrossRefPubMedPubMedCentral
Zurück zum Zitat Clausen J, Vergeiner B, Enk M, Petzer AL, Gastl G, Gunsilius E (2003) Functional significance of the activation-associated receptors CD25 and CD69 on human NK-cells and NK-like T-cells. Immunobiology 207:85–93CrossRefPubMed Clausen J, Vergeiner B, Enk M, Petzer AL, Gastl G, Gunsilius E (2003) Functional significance of the activation-associated receptors CD25 and CD69 on human NK-cells and NK-like T-cells. Immunobiology 207:85–93CrossRefPubMed
Zurück zum Zitat Cooper MA, Fehniger TA, Caligiuri MA (2001) The biology of human natural killer-cell subsets. Trends Immunol 22:633–640CrossRefPubMed Cooper MA, Fehniger TA, Caligiuri MA (2001) The biology of human natural killer-cell subsets. Trends Immunol 22:633–640CrossRefPubMed
Zurück zum Zitat Costello RT, Sivori S, Marcenaro E, Lafage-Pochitaloff M, Mozziconacci MJ, Reviron D, Gastaut JA, Pende D, Olive D, Moretta A (2002) Defective expression and function of natural killer cell-triggering receptors in patients with acute myeloid leukemia. Blood 99:3661–3667CrossRefPubMed Costello RT, Sivori S, Marcenaro E, Lafage-Pochitaloff M, Mozziconacci MJ, Reviron D, Gastaut JA, Pende D, Olive D, Moretta A (2002) Defective expression and function of natural killer cell-triggering receptors in patients with acute myeloid leukemia. Blood 99:3661–3667CrossRefPubMed
Zurück zum Zitat de Rham C, Ferrari-Lacraz S, Jendly S, Schneiter G, Dayer JM, Villard J (2007) The proinflammatory cytokines IL-2, IL-15 and IL-21 modulate the repertoire of mature human natural killer cell receptors. Arthritis Res Ther 9:R125CrossRefPubMedPubMedCentral de Rham C, Ferrari-Lacraz S, Jendly S, Schneiter G, Dayer JM, Villard J (2007) The proinflammatory cytokines IL-2, IL-15 and IL-21 modulate the repertoire of mature human natural killer cell receptors. Arthritis Res Ther 9:R125CrossRefPubMedPubMedCentral
Zurück zum Zitat Dons’koi BV, Chernyshov VP, Osypchuk DV (2011) Measurement of NK activity in whole blood by the CD69 up-regulation after co-incubation with K562, comparison with NK cytotoxicity assays and CD107a degranulation assay. J Immunol Methods 372:187–195CrossRefPubMed Dons’koi BV, Chernyshov VP, Osypchuk DV (2011) Measurement of NK activity in whole blood by the CD69 up-regulation after co-incubation with K562, comparison with NK cytotoxicity assays and CD107a degranulation assay. J Immunol Methods 372:187–195CrossRefPubMed
Zurück zum Zitat Farag SS, Caligiuri MA (2006) Immunologic approaches to acute leukemia in the elderly. Semin Hematol 43:118–125CrossRefPubMed Farag SS, Caligiuri MA (2006) Immunologic approaches to acute leukemia in the elderly. Semin Hematol 43:118–125CrossRefPubMed
Zurück zum Zitat Fauriat C, Just-Landi S, Mallet F, Arnoulet C, Sainty D, Olive D, Costello RT (2007) 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 109:323–330CrossRefPubMed Fauriat C, Just-Landi S, Mallet F, Arnoulet C, Sainty D, Olive D, Costello RT (2007) 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 109:323–330CrossRefPubMed
Zurück zum Zitat Ferrara F, Palmieri S, Mele G (2004) Prognostic factors and therapeutic options for relapsed or refractory acute myeloid leukemia. Haematologica 89:998–1008PubMed Ferrara F, Palmieri S, Mele G (2004) Prognostic factors and therapeutic options for relapsed or refractory acute myeloid leukemia. Haematologica 89:998–1008PubMed
Zurück zum Zitat Garcia-Iglesias T, Del Toro-Arreola A, Albarran-Somoza B, Del Toro-Arreola S, Sanchez-Hernandez PE, Ramirez-Dueñas MG, Balderas-Peña LM, Bravo-Cuellar A, Ortiz-Lazareno PC, Daneri-Navarro A (2009) Low NKp30, NKp46 and NKG2D expression and reduced cytotoxic activity on NK cells in cervical cancer and precursor lesions. BMC Cancer 9:186CrossRefPubMedPubMedCentral Garcia-Iglesias T, Del Toro-Arreola A, Albarran-Somoza B, Del Toro-Arreola S, Sanchez-Hernandez PE, Ramirez-Dueñas MG, Balderas-Peña LM, Bravo-Cuellar A, Ortiz-Lazareno PC, Daneri-Navarro A (2009) Low NKp30, NKp46 and NKG2D expression and reduced cytotoxic activity on NK cells in cervical cancer and precursor lesions. BMC Cancer 9:186CrossRefPubMedPubMedCentral
Zurück zum Zitat Huergo-Zapico L, Acebes-Huerta A, Gonzalez-Rodriguez AP, Contesti J, Gonzalez-García E, Payer AR, Villa-Alvarez M, Fernández-Guizán A, López-Soto A, Gonzalez S (2014) Expansion of NK cells and reduction of NKG2D expression in chronic lymphocytic leukemia. Correlation with progressive disease. PLoS ONE 9:e108326CrossRefPubMedPubMedCentral Huergo-Zapico L, Acebes-Huerta A, Gonzalez-Rodriguez AP, Contesti J, Gonzalez-García E, Payer AR, Villa-Alvarez M, Fernández-Guizán A, López-Soto A, Gonzalez S (2014) Expansion of NK cells and reduction of NKG2D expression in chronic lymphocytic leukemia. Correlation with progressive disease. PLoS ONE 9:e108326CrossRefPubMedPubMedCentral
Zurück zum Zitat Karre K (2002) NK cells, MHC class I molecules and the missing self. Scand J Immunol 55:221–228CrossRefPubMed Karre K (2002) NK cells, MHC class I molecules and the missing self. Scand J Immunol 55:221–228CrossRefPubMed
Zurück zum Zitat Kern W, Danhauser-Riedl S, Ratei R, Schnittger S, Schoch C, Kolb HJ, Ludwig WD, Hiddemann W, Haferlach T (2003) Detection of minimal residual disease in unselected patients with acute myeloid leukemia using multiparameter flow cytometry for definition of leukemia-associated immunophenotypes and determination of their frequencies in normal bone marrow. Haematologica 88:646–653PubMed Kern W, Danhauser-Riedl S, Ratei R, Schnittger S, Schoch C, Kolb HJ, Ludwig WD, Hiddemann W, Haferlach T (2003) Detection of minimal residual disease in unselected patients with acute myeloid leukemia using multiparameter flow cytometry for definition of leukemia-associated immunophenotypes and determination of their frequencies in normal bone marrow. Haematologica 88:646–653PubMed
Zurück zum Zitat Kim EK, Ahn YO, Kim S, Kim TM, Keam B, Heo DS (2013) Ex vivo activation and expansion of natural killer cells from patients with advanced cancer with feeder cells from healthy volunteers. Cytotherapy 15(231–241):e1 Kim EK, Ahn YO, Kim S, Kim TM, Keam B, Heo DS (2013) Ex vivo activation and expansion of natural killer cells from patients with advanced cancer with feeder cells from healthy volunteers. Cytotherapy 15(231–241):e1
Zurück zum Zitat Liao W, Lin JX, Leonard WJ (2011) IL-2 family cytokines: new insights into the complex roles of IL-2 as a broad regulator of T helper cell differentiation. Curr Opin Immunol 23:598–604CrossRefPubMedPubMedCentral Liao W, Lin JX, Leonard WJ (2011) IL-2 family cytokines: new insights into the complex roles of IL-2 as a broad regulator of T helper cell differentiation. Curr Opin Immunol 23:598–604CrossRefPubMedPubMedCentral
Zurück zum Zitat Luhm J, Brand JM, Koritke P, Hoppner M, Kirchner H, Frohn C (2002) Large-scale generation of natural killer lymphocytes for clinical application. J Hematother Stem Cell Res 11:651–657CrossRefPubMed Luhm J, Brand JM, Koritke P, Hoppner M, Kirchner H, Frohn C (2002) Large-scale generation of natural killer lymphocytes for clinical application. J Hematother Stem Cell Res 11:651–657CrossRefPubMed
Zurück zum Zitat Melder RJ, Walker ER, Herberman RB, Whiteside TL (1990) Surface characteristics, morphology, and ultrastructure of human adherent lymphokine-activated killer cells. J Leukoc Biol 48:163–173PubMed Melder RJ, Walker ER, Herberman RB, Whiteside TL (1990) Surface characteristics, morphology, and ultrastructure of human adherent lymphokine-activated killer cells. J Leukoc Biol 48:163–173PubMed
Zurück zum Zitat Moretta L, Moretta A (2004) Unravelling natural killer cell function: triggering and inhibitory human NK receptors. EMBO J 23:255–259CrossRefPubMed Moretta L, Moretta A (2004) Unravelling natural killer cell function: triggering and inhibitory human NK receptors. EMBO J 23:255–259CrossRefPubMed
Zurück zum Zitat Rodella L, Rezzani R, Zauli G, Mariani AR, Rizzoli R, Vitale M (1998) Apoptosis induced by NK cells is modulated by the NK-active cytokines IL-2 and IL-12. Int Immunol 10:719–725CrossRefPubMed Rodella L, Rezzani R, Zauli G, Mariani AR, Rizzoli R, Vitale M (1998) Apoptosis induced by NK cells is modulated by the NK-active cytokines IL-2 and IL-12. Int Immunol 10:719–725CrossRefPubMed
Zurück zum Zitat Rodella L, Zamai L, Rezzani R, Artico M, Peri G, Falconi M, Facchini A, Pelusi G, Vitale M (2001) Interleukin 2 and interleukin 15 differentially predispose natural killer cells to apoptosis mediated by endothelial and tumour cells. Br J Haematol 115:442–450CrossRefPubMed Rodella L, Zamai L, Rezzani R, Artico M, Peri G, Falconi M, Facchini A, Pelusi G, Vitale M (2001) Interleukin 2 and interleukin 15 differentially predispose natural killer cells to apoptosis mediated by endothelial and tumour cells. Br J Haematol 115:442–450CrossRefPubMed
Zurück zum Zitat Ruggeri L, Mancusi A, Capanni M, Urbani E, Carotti A, Aloisi T, Stern M, Pende D, Perruccio K, Burchielli E, Topini F, Bianchi E, Aversa F, Martelli MF, Velardi A (2007) Donor natural killer cell allorecognition of missing self in haploidentical hematopoietic transplantation for acute myeloid leukemia: challenging its predictive value. Blood 110:433–440CrossRefPubMedPubMedCentral Ruggeri L, Mancusi A, Capanni M, Urbani E, Carotti A, Aloisi T, Stern M, Pende D, Perruccio K, Burchielli E, Topini F, Bianchi E, Aversa F, Martelli MF, Velardi A (2007) Donor natural killer cell allorecognition of missing self in haploidentical hematopoietic transplantation for acute myeloid leukemia: challenging its predictive value. Blood 110:433–440CrossRefPubMedPubMedCentral
Zurück zum Zitat Saito H, Osaki T, Ikeguchi M (2012) Decreased NKG2D expression on NK cells correlates with impaired NK cell function in patients with gastric cancer. Gastric Cancer 15:27–33CrossRefPubMed Saito H, Osaki T, Ikeguchi M (2012) Decreased NKG2D expression on NK cells correlates with impaired NK cell function in patients with gastric cancer. Gastric Cancer 15:27–33CrossRefPubMed
Zurück zum Zitat Salih HR, Rammensee HG, Steinle A (2002) Cutting edge: down-regulation of MICA on human tumors by proteolytic shedding. J Immunol 169:4098–4102CrossRefPubMed Salih HR, Rammensee HG, Steinle A (2002) Cutting edge: down-regulation of MICA on human tumors by proteolytic shedding. J Immunol 169:4098–4102CrossRefPubMed
Zurück zum Zitat Schnittger S, Weisser M, Schoch C, Hiddemann W, Haferlach T, Kern W (2003) New score predicting for prognosis in PML-RARA+, AML1-ETO+, or CBFBMYH11+ acute myeloid leukemia based on quantification of fusion transcripts. Blood 102:2746–2755CrossRefPubMed Schnittger S, Weisser M, Schoch C, Hiddemann W, Haferlach T, Kern W (2003) New score predicting for prognosis in PML-RARA+, AML1-ETO+, or CBFBMYH11+ acute myeloid leukemia based on quantification of fusion transcripts. Blood 102:2746–2755CrossRefPubMed
Zurück zum Zitat Trapani JA, Smyth MJ (2002) Functional significance of the perforin/granzyme cell death pathway. Nat Rev Immunol 2:735–747CrossRefPubMed Trapani JA, Smyth MJ (2002) Functional significance of the perforin/granzyme cell death pathway. Nat Rev Immunol 2:735–747CrossRefPubMed
Zurück zum Zitat Vitale M, Bottino C, Sivori S, Sanseverino L, Castriconi R, Marcenaro E, Augugliaro R, Moretta L, Moretta A (1998) NKp44, a novel triggering surface molecule specifically expressed by activated natural killer cells, is involved in non-major histocompatibility complex-restricted tumor cell lysis. J Exp Med 187:2065–2072CrossRefPubMedPubMedCentral Vitale M, Bottino C, Sivori S, Sanseverino L, Castriconi R, Marcenaro E, Augugliaro R, Moretta L, Moretta A (1998) NKp44, a novel triggering surface molecule specifically expressed by activated natural killer cells, is involved in non-major histocompatibility complex-restricted tumor cell lysis. J Exp Med 187:2065–2072CrossRefPubMedPubMedCentral
Zurück zum Zitat Vujanovic NL, Rabinowich H, Lee YJ, Jost L, Herberman RB, Whiteside TL (1993) Distinct phenotypic and functional characteristics of human natural killer cells obtained by rapid interleukin 2-induced adherence to plastic. Cell Immunol 151:133–157CrossRefPubMed Vujanovic NL, Rabinowich H, Lee YJ, Jost L, Herberman RB, Whiteside TL (1993) Distinct phenotypic and functional characteristics of human natural killer cells obtained by rapid interleukin 2-induced adherence to plastic. Cell Immunol 151:133–157CrossRefPubMed
Zurück zum Zitat Waldhauer I, Steinle A (2008) NK cells and cancer immunosurveillance. Oncogene 27:5932–5943CrossRefPubMed Waldhauer I, Steinle A (2008) NK cells and cancer immunosurveillance. Oncogene 27:5932–5943CrossRefPubMed
Zurück zum Zitat Wallin RP, Screpanti V, Michaelsson J, Grandien A, Ljunggren HG (2003) Regulation of perforin-independent NK cell-mediated cytotoxicity. Eur J Immunol 33:2727–2735CrossRefPubMed Wallin RP, Screpanti V, Michaelsson J, Grandien A, Ljunggren HG (2003) Regulation of perforin-independent NK cell-mediated cytotoxicity. Eur J Immunol 33:2727–2735CrossRefPubMed
Zurück zum Zitat Yawata M, Yawata N, Draghi M, Little AM, Partheniou F, Parham P (2006) Roles for HLA and KIR polymorphisms in natural killer cell repertoire selection and modulation of effector function. J Exp Med 203:633–645CrossRefPubMedPubMedCentral Yawata M, Yawata N, Draghi M, Little AM, Partheniou F, Parham P (2006) Roles for HLA and KIR polymorphisms in natural killer cell repertoire selection and modulation of effector function. J Exp Med 203:633–645CrossRefPubMedPubMedCentral
Metadaten
Titel
Cryopreserved NK cells in the treatment of haematological malignancies: preclinical study
verfasst von
Monika Holubova
Michaela Miklikova
Martin Leba
Daniel Georgiev
Pavel Jindra
Martin Caprnda
Rachele Ciccocioppo
Peter Kruzliak
Daniel Lysak
Publikationsdatum
10.09.2016
Verlag
Springer Berlin Heidelberg
Erschienen in
Journal of Cancer Research and Clinical Oncology / Ausgabe 12/2016
Print ISSN: 0171-5216
Elektronische ISSN: 1432-1335
DOI
https://doi.org/10.1007/s00432-016-2247-8

Weitere Artikel der Ausgabe 12/2016

Journal of Cancer Research and Clinical Oncology 12/2016 Zur Ausgabe

Update Onkologie

Bestellen Sie unseren Fach-Newsletter und bleiben Sie gut informiert.