Cancer Immunology, Immunotherapy

, Volume 65, Issue 4, pp 485–492 | Cite as

NK-92: an ‘off-the-shelf therapeutic’ for adoptive natural killer cell-based cancer immunotherapy

  • Garnet Suck
  • Marcus Odendahl
  • Paulina Nowakowska
  • Christian Seidl
  • Winfried S. Wels
  • Hans G. Klingemann
  • Torsten TonnEmail author
Symposium-in-writing paper


Natural killer (NK) cells are increasingly considered as immunotherapeutic agents in particular in the fight against cancers. NK cell therapies are potentially broadly applicable and, different from their T cell counterparts, do not cause graft-versus-host disease. Efficacy and clinical in vitro or in vivo expansion of primary NK cells will however always remain variable due to individual differences of donors or patients. Long-term storage of clinical NK cell lots to allow repeated clinical applications remains an additional challenge. In contrast, the established and well-characterized cell line NK-92 can be easily and reproducibly expanded from a good manufacturing practice (GMP)-compliant cryopreserved master cell bank. Moreover, no cost-intensive cell purification methods are required. To date, NK-92 has been intensively studied. The cells displayed superior cytotoxicity against a number of tumor types tested, which was confirmed in preclinical mouse studies. Subsequent clinical testing demonstrated safety of NK-92 infusions even at high doses. Despite the phase I nature of the trials conducted so far, some efficacy was noted, particularly against lung tumors. Furthermore, to overcome tumor resistance and for specific targeting, NK-92 has been engineered to express a number of different chimeric antigen receptors (CARs), including targeting, for example, CD19 or CD20 (anti-B cell malignancies), CD38 (anti-myeloma) or human epidermal growth factor receptor 2 (HER2; ErbB2; anti-epithelial cancers). The concept of an NK cell line as an allogeneic cell therapeutic produced ‘off-the-shelf’ on demand holds great promise for the development of effective treatments.


NK-92 Cellular immunotherapy CAR Clinical trial NK cell line Tumor targeting 



Antibody-dependent cell-mediated cytotoxicity


Acute lymphoblastic leukemia


Bi-specific or tri-specific killer engager


Chimeric antigen receptor


Chronic lymphocytic leukemia


Epstein–Barr virus


Epithelial cell adhesion molecule




Fragment crystallizable region


Good manufacturing practice


Graft-versus-host disease

HER2; ErbB2

Human epidermal growth factor receptor 2


Human leukocyte antigen


Immunoglobulin G




Killer cell immunoglobulin-like receptor


Leukocyte immunoglobulin-like receptor


MHC class I chain-related gene A


MHC class I chain-related gene B


Natural killer


Natural killer group


Peripheral blood mononuclear cell


Renal cell carcinoma


Response Evaluation Criteria in Solid Tumors


Single-chain variable fragment


Severe combined immunodeficiency


Compliance with ethical standards

Conflict of interest

Dr. H. G. Klingemann is affiliated with NantKwest Inc, CA, USA (formerly known as Conkwest, Inc.). All other authors declare no conflicts of interest.


  1. 1.
    Klingemann HG, Martinson J (2004) Ex vivo expansion of natural killer cells for clinical applications. Cytotherapy 6:15–22CrossRefPubMedGoogle Scholar
  2. 2.
    Suck G, Koh MB (2010) Emerging natural killer cell immunotherapies: large-scale ex vivo production of highly potent anticancer effectors. Hematol Oncol Stem Cell Ther 3:135–142CrossRefPubMedGoogle Scholar
  3. 3.
    Childs RW, Berg M (2013) Bringing natural killer cells to the clinic: ex vivo manipulation. Hematol Am Soc Hematol Educ Progr 2013:234–246CrossRefGoogle Scholar
  4. 4.
    Gong JH, Maki G, Klingemann HG (1994) Characterization of a human cell line (nk-92) with phenotypical and functional characteristics of activated natural killer cells. Leukemia 8:652–658PubMedGoogle Scholar
  5. 5.
    Tam YK, Martinson JA, Doligosa K, Klingemann HG (2003) Ex vivo expansion of the highly cytotoxic human natural killer-92 cell-line under current good manufacturing practice conditions for clinical adoptive cellular immunotherapy. Cytotherapy 5:259–272CrossRefPubMedGoogle Scholar
  6. 6.
    Tonn T, Becker S, Esser R, Schwabe D, Seifried E (2001) Cellular immunotherapy of malignancies using the clonal natural killer cell line NK-92. J Hematother Stem Cell Res 10:535–544CrossRefPubMedGoogle Scholar
  7. 7.
    Maki G, Klingemann HG, Martinson JA, Tam YK (2001) Factors regulating the cytotoxic activity of the human natural killer cell line, nk-92. J Hematother Stem Cell Res 10:369–383CrossRefPubMedGoogle Scholar
  8. 8.
    Luetke-Eversloh M, Killig M, Romagnani C (2013) Signatures of human nk cell development and terminal differentiation. Front Immunol 4:499CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Matsuo Y, Drexler HG (2003) Immunoprofiling of cell lines derived from natural killer-cell and natural killer-like t-cell leukemia-lymphoma. Leuk Res 27:935–945CrossRefPubMedGoogle Scholar
  10. 10.
    Cooley S, Xiao F, Pitt M, Gleason M, McCullar V, Bergemann TL, McQueen KL, Guethlein LA, Parham P, Miller JS (2007) A subpopulation of human peripheral blood nk cells that lacks inhibitory receptors for self-mhc is developmentally immature. Blood 110:578–586CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Faure M, Long EO (2002) Kir2dl4 (cd158d), an nk cell-activating receptor with inhibitory potential. J Immunol 168:6208–6214CrossRefPubMedGoogle Scholar
  12. 12.
    Pazmany L, Mandelboim O, Vales-Gomez M, Davis DM, Reyburn HT, Strominger JL (1996) Protection from natural killer cell-mediated lysis by hla-g expression on target cells. Science 274:792–795CrossRefPubMedGoogle Scholar
  13. 13.
    Romanski A, Bug G, Becker S, Kampfmann M, Seifried E, Hoelzer D, Ottmann OG, Tonn T (2005) Mechanisms of resistance to natural killer cell-mediated cytotoxicity in acute lymphoblastic leukemia. Exp Hematol 33:344–352CrossRefPubMedGoogle Scholar
  14. 14.
    Burshtyn DN, Scharenberg AM, Wagtmann N, Rajagopalan S, Berrada K, Yi T, Kinet JP, Long EO (1996) Recruitment of tyrosine phosphatase hcp by the killer cell inhibitor receptor. Immunity 4:77–85CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Heidenreich S, ZuEulenburg C, Hildebrandt Y, Stubig T, Sierich H, Badbaran A, Eiermann TH, Binder TM, Kroger N (2012) Impact of the nk cell receptor lir-1 (ilt-2/cd85j/lilrb1) on cytotoxicity against multiple myeloma. Clin Dev immunol. 2012:652130CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Suck G, Branch DR, Smyth MJ, Miller RG, Vergidis J, Fahim S, Keating A (2005) Khyg-1, a model for the study of enhanced natural killer cell cytotoxicity. Exp Hematol 33:1160–1171CrossRefPubMedGoogle Scholar
  17. 17.
    Gong JH, Maki G, Klingemann HG (1994) Characterization of a human cell line (nk-92) with phenotypical and functional characteristics of activated natural killer cells. Leukemia 8:652–658PubMedGoogle Scholar
  18. 18.
    Klingemann HG, Wong E, Maki G (1996) A cytotoxic nk-cell line (nk-92) for ex vivo purging of leukemia from blood. Biol Blood Marrow Transplant Biol Blood Marrow Transplant 2:68–75PubMedGoogle Scholar
  19. 19.
    Yan Y, Steinherz P, Klingemann HG, Dennig D, Childs BH, McGuirk J, O’Reilly RJ (1998) Antileukemia activity of a natural killer cell line against human leukemias. Clin Cancer Res 4:2859–2868PubMedGoogle Scholar
  20. 20.
    Lowdell MW, Theocharous P (1997) “Less is more”: the role of purging in hematopoietic stem cell transplantation. Oncologist 2:268–274PubMedGoogle Scholar
  21. 21.
    Yahng SA, Yoon JH, Shin SH, Lee SE, Cho BS, Eom KS, Kim YJ, Lee S, Kim HJ, Min CK, Kim DW, Lee JW, Min WS, Park CW, Kim Y, Cho SG (2014) Influence of ex vivo purging with clinimacs cd34(+) selection on outcome after autologous stem cell transplantation in non-Hodgkin lymphoma. Br J Haematol 164:555–564CrossRefPubMedGoogle Scholar
  22. 22.
    Bais S, Bartee E, Rahman MM, McFadden G, Cogle CR (2012) Oncolytic virotherapy for hematological malignancies. Adv Virol 2012:186512CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Maki G, Tam YK, Berkahn L, Klingemann HG (2003) Ex vivo purging with nk-92 prior to autografting for chronic myelogenous leukemia. Bone Marrow Transplant 31:1119–1125CrossRefPubMedGoogle Scholar
  24. 24.
    Maki G (2001) Ex vivo purging of stem cell autografts using cytotoxic cells. J Hematother Stem Cell Res 10:545–551CrossRefPubMedGoogle Scholar
  25. 25.
    Klingemann HG (2013) Cellular therapy of cancer with natural killer cells-where do we stand? Cytotherapy 15:1185–1194CrossRefPubMedGoogle Scholar
  26. 26.
    Tonn T, Seifried E (2006) Natural killer cells for the treatment of malignancies. Transfus Med Hemother 33:144–149CrossRefGoogle Scholar
  27. 27.
    Tonn T, Schwabe D, Klingemann HG, Becker S, Esser R, Koehl U, Suttorp M, Seifried E, Ottmann OG, Bug G (2013) Treatment of patients with advanced cancer with the natural killer cell line NK-92. Cytotherapy 15:1563–1570CrossRefPubMedGoogle Scholar
  28. 28.
    Arai S, Meagher R, Swearingen M, Myint H, Rich E, Martinson J, Klingemann H (2008) Infusion of the allogeneic cell line nk-92 in patients with advanced renal cell cancer or melanoma: a phase i trial. Cytotherapy 10:625–632CrossRefPubMedGoogle Scholar
  29. 29.
    Tam YK, Miyagawa B, Ho VC, Klingemann HG (1999) Immunotherapy of malignant melanoma in a scid mouse model using the highly cytotoxic natural killer cell line nk-92. J Hematother 8:281–290CrossRefPubMedGoogle Scholar
  30. 30.
    Whiteside TL, Griffin DL, Stanson J, Gooding W, McKenna D, Sumstad D, Kadidlo D, Gee A, Durett A, Lindblad R, Wood D, Styers D (2011) Shipping of therapeutic somatic cell products. Cytotherapy 13:201–213CrossRefPubMedGoogle Scholar
  31. 31.
    Koepsell SA, Kadidlo DM, Fautsch S, McCullough J, Klingemann H, Wagner JE, Miller JS, McKenna DH Jr (2013) Successful “in-flight” activation of natural killer cells during long-distance shipping. Transfusion 53:398–403CrossRefPubMedGoogle Scholar
  32. 32.
    Suck G, Branch DR, Aravena P, Mathieson M, Helke S, Keating A (2006) Constitutively polarized granules prime khyg-1 nk cells. Int Immunol 18:1347–1354CrossRefPubMedGoogle Scholar
  33. 33.
    Suck G, Branch DR, Keating A (2006) Irradiated khyg-1 retains cytotoxicity: Potential for adoptive immunotherapy with a natural killer cell line. Int J Radiat Biol 82:355–361CrossRefPubMedGoogle Scholar
  34. 34.
    Suck G, Tan SM, Chu S, Niam M, Vararattanavech A, Lim TJ, Koh MB (2011) Khyg-1 and nk-92 represent different subtypes of lfa-1-mediated nk cell adhesiveness. Front Biosci 3:166–178CrossRefGoogle Scholar
  35. 35.
    Mallett CL, McFadden C, Chen Y, Foster PJ (2012) Migration of iron-labeled khyg-1 natural killer cells to subcutaneous tumors in nude mice, as detected by magnetic resonance imaging. Cytotherapy 14:743–751CrossRefPubMedGoogle Scholar
  36. 36.
    Swift BE, Williams BA, Kosaka Y, Wang XH, Medin JA, Viswanathan S, Martinez-Lopez J, Keating A (2012) Natural killer cell lines preferentially kill clonogenic multiple myeloma cells and decrease myeloma engraftment in a bioluminescent xenograft mouse model. Haematologica 97:1020–1028CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Yagita M, Huang CL, Umehara H, Matsuo Y, Tabata R, Miyake M, Konaka Y, Takatsuki K (2000) A novel natural killer cell line (khyg-1) from a patient with aggressive natural killer cell leukemia carrying a p53 point mutation. Leukemia 14:922–930CrossRefPubMedGoogle Scholar
  38. 38.
    Porter DL, Levine BL, Kalos M, Bagg A, June CH (2011) Chimeric antigen receptor-modified t cells in chronic lymphoid leukemia. N Engl J Med 365:725–733CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Wieczorek A, Uharek L (2013) Genetically modified t cells for the treatment of malignant disease. Transfus Med Hemother 40:388–402CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Bhat R, Watzl C (2007) Serial killing of tumor cells by human natural killer cells–enhancement by therapeutic antibodies. PLoS ONE 2:e326CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Choi PJ, Mitchison TJ (2013) Imaging burst kinetics and spatial coordination during serial killing by single natural killer cells. Proc Natl Acad Sci USA 110:6488–6493CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Boissel L, Betancur M, Lu W, Wels WS, Marino T, Van Etten RA, Klingemann H (2012) Comparison of mRNA and lentiviral based transfection of natural killer cells with chimeric antigen receptors recognizing lymphoid antigens. Leuk Lymphoma 53:958–965CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Suck G (2006) Novel approaches using natural killer cells in cancer therapy. Semin Cancer Biol 16:412–418CrossRefPubMedGoogle Scholar
  44. 44.
    Boissel L, Betancur M, Wels WS, Tuncer H, Klingemann H (2009) Transfection with mrna for cd19 specific chimeric antigen receptor restores nk cell mediated killing of cll cells. Leuk Res 33:1255–1259CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Muller T, Uherek C, Maki G, Chow KU, Schimpf A, Klingemann HG, Tonn T, Wels WS (2008) Expression of a cd20-specific chimeric antigen receptor enhances cytotoxic activity of nk cells and overcomes nk-resistance of lymphoma and leukemia cells. Cancer Immunol Immunother 57:411–423CrossRefPubMedGoogle Scholar
  46. 46.
    Boissel L, Betancur-Boissel M, Lu W, Krause DS, Van Etten RA, Wels WS, Klingemann H (2013) Retargeting nk-92 cells by means of cd19- and cd20-specific chimeric antigen receptors compares favorably with antibody-dependent cellular cytotoxicity. Oncoimmunology 2:e26527CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Miller JS (2013) Therapeutic applications: Natural killer cells in the clinic. Hematol Am Soc Hematol Educ Progr 2013:247–253CrossRefGoogle Scholar
  48. 48.
    Uherek C, Tonn T, Uherek B, Becker S, Schnierle B, Klingemann HG, Wels W (2002) Retargeting of natural killer-cell cytolytic activity to erbb2-expressing cancer cells results in efficient and selective tumor cell destruction. Blood 100:1265–1273PubMedGoogle Scholar
  49. 49.
    Daldrup-Link HE, Meier R, Rudelius M, Piontek G, Piert M, Metz S, Settles M, Uherek C, Wels W, Schlegel J, Rummeny EJ (2005) In vivo tracking of genetically engineered, anti-her2/neu directed natural killer cells to her2/neu positive mammary tumors with magnetic resonance imaging. Eur Radiol 15:4–13CrossRefPubMedGoogle Scholar
  50. 50.
    Meier R, Piert M, Piontek G, Rudelius M, Oostendorp RA, Senekowitsch-Schmidtke R, Henning TD, Wels WS, Uherek C, Rummeny EJ, Daldrup-Link HE (2008) Tracking of [18f]fdg-labeled natural killer cells to her2/neu-positive tumors. Nucl Med Biol 35:579–588CrossRefPubMedGoogle Scholar
  51. 51.
    Schönfeld K, Sahm C, Zhang C, Naundorf S, Brendel C, Odendahl M, Nowakowska P, Bönig H, Köhl U, Kloess S, Köhler S, Holtgreve-Grez H, Jauch A, Schmidt M, Schubert R, Kühlcke K, Seifried E, Klingemann HG, Rieger MA, Tonn T, Grez M, Wels WS (2015) Selective inhibition of tumor growth by NK cells expressing an ErbB2/HER2-specific chimeric antigen receptor. Mol Ther 23:330–338CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Tam YK, Maki G, Miyagawa B, Hennemann B, Tonn T, Klingemann HG (1999) Characterization of genetically altered, interleukin 2-independent natural killer cell lines suitable for adoptive cellular immunotherapy. Hum Gene Ther 10:1359–1373CrossRefPubMedGoogle Scholar
  53. 53.
    Reid GS, Bharya S, Klingemann HG, Schultz KR (2002) Differential killing of pre-b acute lymphoblastic leukaemia cells by activated nk cells and the nk-92 ci cell line. Clin Exp Immunol 129:265–271CrossRefPubMedPubMedCentralGoogle Scholar
  54. 54.
    Sahm C, Schonfeld K, Wels WS (2012) Expression of il-15 in nk cells results in rapid enrichment and selective cytotoxicity of gene-modified effectors that carry a tumor-specific antigen receptor. Cancer Immunol Immunother 61:1451–1461CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Garnet Suck
    • 1
  • Marcus Odendahl
    • 2
  • Paulina Nowakowska
    • 3
  • Christian Seidl
    • 3
  • Winfried S. Wels
    • 4
  • Hans G. Klingemann
    • 5
  • Torsten Tonn
    • 2
    • 3
    • 6
    Email author
  1. 1.Institute for Transfusion MedicineGerman Red Cross Blood Donation Service North-EastBerlinGermany
  2. 2.Institute for Transfusion MedicineGerman Red Cross Blood Donation Service North-EastDresdenGermany
  3. 3.Institute for Transfusion Medicine and ImmunohematologyGerman Red Cross Blood Donation Service Baden-Württemberg-HessenFrankfurt am MainGermany
  4. 4.Institute for Tumor Biology and Experimental TherapyGeorg-Speyer-HausFrankfurt am MainGermany
  5. 5.NantKwest IncCulver CityUSA
  6. 6.Medical Faculty Carl Gustav CarusTechnische Universität DresdenDresdenGermany

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