Lytic activity against primary AML cells is stimulated in vitro by an autologous whole cell vaccine expressing IL-2 and CD80

  • Nicola Hardwick
  • Lucas Chan
  • Wendy Ingram
  • Ghulam Mufti
  • Farzin FarzanehEmail author
Original Article


Despite being of the myeloid lineage, acute myeloid leukaemia (AML) blasts are of low immunogenicity, probably because they lack the costimulatory molecule CD80 and secrete immunosuppressive factors. We have previously shown that in vitro stimulation of autologous peripheral blood mononuclear cells (PBMCs) with primary AML cells modified to express CD80 and IL-2 promotes proliferation, secretion of Th1 cytokines and expansion of activated CD8+ T cells. In this study, we show that allogeneic effector cells (from a healthy donor or AML patients) when stimulated with IL-2/CD80 modified AML blasts were able to induce the lysis of unmodified AML blasts. Effector cells stimulated with IL-2/CD80AML blasts had higher lytic activity than cells stimulated with AML cells expressing CD80 or IL-2 alone. Similarly, AML patient PBMCs primed with autologous IL-2/CD80 AML cells had a higher frequency of IFN-γ secreting cells and show cytotoxicity against autologous, unmodified blasts. Crucially, the response appears to be leukaemia specific, since stimulated patient PBMCs show higher frequencies of IFN-γ secreting effector cells in response to AML blasts than to remission bone marrow cells from the same patients. Although studied in a small number of heterogeneous patient samples, the data are encouraging and support the continuing development of vaccination for poor prognosis AML patients with autologous cells genetically modified to express IL-2/CD80.


Acute myeloid leukaemia Cellular therapy Gene therapy CD80 IL-2 



We are grateful to Dr Linda Barber for critical feedback on this manuscript. This work was funded by The Leukaemia Research Fund and The Elimination of Leukaemia Fund. The authors also acknowledge financial support from the Department of Health via the National Institute for Health Research (NIHR) comprehensive Biomedical Research Centre award to Guy’s and St Thomas’ NHS Foundation Trust in partnership with King’s College London.


  1. 1.
    Goldstone AH, Burnett AK, Wheatley K, Smith AG, Hutchinson RM, Clark RE (2001) Attempts to improve treatment outcomes in acute myeloid leukaemia (AML) in older patients: the results of the United Kingdom Medical Research Council AML11 trial. Blood 98:1302CrossRefPubMedGoogle Scholar
  2. 2.
    Hann IM, Stevens RF, Goldstone AH, Rees JK, Wheatley K, Gray RG, Burnett AK (1997) Randomized comparison of DAT versus ADE as induction chemotherapy in children and younger adults with acute myeloid leukaemia. Results of the Medical Research Council’s 10th AML trial (MRC AML10). Adult and Childhood Leukaemia Working Parties of the Medical Research Council. Blood 89:2311PubMedGoogle Scholar
  3. 3.
    Chan L, Hardwick NR, Guinn BA, Darling D, Gaken J, Galea-Lauri J, Ho AY, Mufti GJ, Farzaneh F (2006) An immune edited tumour versus a tumour edited immune system: prospects for immune therapy of acute myeloid leukaemia. Cancer Immunol Immunother 55:1017CrossRefPubMedGoogle Scholar
  4. 4.
    Molldrem JJ (2006) Vaccination for leukaemia. Biol Blood Marrow Transplant 12:13CrossRefPubMedGoogle Scholar
  5. 5.
    Kolb HJ, Schmid C, Barrett AJ, Schendel DJ (2004) Graft-versus-leukemia reactions in allogeneic chimeras. Blood 103:767CrossRefPubMedGoogle Scholar
  6. 6.
    Andersen MH, Svane IM, Kvistborg P, Nielsen OJ, Balslev E, Reker S, Becker JC, Straten PT (2005) Immunogenicity of Bcl-2 in patients with cancer. Blood 105:728CrossRefPubMedGoogle Scholar
  7. 7.
    Greiner J, Ringhoffer M, Taniguchi M, Schmitt A, Kirchner D, Krahn G, Heilmann V, Gschwend J, Bergmann L, Dohner H, Schmitt M (2002) Receptor for hyaluronan acid-mediated motility (RHAMM) is a new immunogenic leukemia-associated antigen in acute and chronic myeloid leukemia. Exp Hematol 30:1029CrossRefPubMedGoogle Scholar
  8. 8.
    Greiner J, Ringhoffer M, Taniguchi M, Hauser T, Schmitt A, Dohner H, Schmitt M (2003) Characterization of several leukaemia-associated antigens inducing humoral immune responses in acute and chronic myeloid leukaemia. Int J Cancer 106:224CrossRefPubMedGoogle Scholar
  9. 9.
    Guinn BA, Gilkes AF, Woodward E, Westwood NB, Mufti GJ, Linch D, Burnett AK, Mills KI (2005) Microarray analysis of tumour antigen expression in presentation acute myeloid leukaemia. Biochem Biophys Res Commun 333:703CrossRefPubMedGoogle Scholar
  10. 10.
    Scheibenbogen C, Letsch A, Thiel E, Schmittel A, Mailaender V, Baerwolf S, Nagorsen D, Keilholz U (2002) CD8 T-cell responses to Wilms tumor gene product WT1 and proteinase 3 in patients with acute myeloid leukaemia. Blood 100:2132CrossRefPubMedGoogle Scholar
  11. 11.
    Lim SH, Worman CP, Jewell AP, Goldstone AH (1991) Cellular cytotoxic function and potential in acute myelogenous leukaemia. Leuk Res 15:641CrossRefPubMedGoogle Scholar
  12. 12.
    Morikawa K, Nakano A, Oseko F, Morikawa S (1989) Depressed natural killer (NK) function in blood and marrow is related to the decrease in CD11+ cells in acute leukaemia. Jpn J Med 28:485PubMedGoogle Scholar
  13. 13.
    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 leukaemia. Blood 99:3661CrossRefPubMedGoogle Scholar
  14. 14.
    Buggins AG, Hirst WJ, Pagliuca A, Mufti GJ (1998) Variable expression of CD3-zeta and associated protein tyrosine kinases in lymphocytes from patients with myeloid malignancies. Br J Haematol 100:784CrossRefPubMedGoogle Scholar
  15. 15.
    Wendelbo O, Nesthus I, Sjo M, Paulsen K, Ernst P, Bruserud O (2004) Functional characterization of T lymphocytes derived from patients with acute myelogenous leukaemia and chemotherapy-induced leukopenia. Cancer Immunol Immunother 53:740CrossRefPubMedGoogle Scholar
  16. 16.
    Mackall CL (2000) T-cell immunodeficiency following cytotoxic antineoplastic therapy: a review. Stem Cells 18:10CrossRefPubMedGoogle Scholar
  17. 17.
    Nguyen S, Dhedin N, Vernant JP, Kuentz M, Al Jijakli A, Rouas-Freiss N, Carosella ED, Boudifa A, Debre P, Vieillard V (2005) NK-cell reconstitution after haploidentical hematopoietic stem-cell transplantations: immaturity of NK cells and inhibitory effect of NKG2A override GvL effect. Blood 105:4135CrossRefPubMedGoogle Scholar
  18. 18.
    Whiteway A, Corbett T, Anderson R, Macdonald I, Prentice HG (2003) Expression of co-stimulatory molecules on acute myeloid leukaemia blasts may effect duration of first remission. Br J Haematol 120:442CrossRefPubMedGoogle Scholar
  19. 19.
    Chamuleau ME, Souwer Y, Van Ham SM, Zevenbergen A, Westers TM, Berkhof J, Meijer CJ, van de Loosdrecht AA, Ossenkoppele GJ (2004) Class II-associated invariant chain peptide expression on myeloid leukemic blasts predicts poor clinical outcome. Cancer Res 64:5546CrossRefPubMedGoogle Scholar
  20. 20.
    Buggins AG, Milojkovic D, Arno MJ, Lea NC, Mufti GJ, Thomas NS, Hirst WJ (2001) Microenvironment produced by acute myeloid leukaemia cells prevents T cell activation and proliferation by inhibition of NF-kappaB, c-Myc, and pRb pathways. J Immunol 167:6021PubMedGoogle Scholar
  21. 21.
    Orleans-Lindsay JK, Barber LD, Prentice HG, Lowdell MW (2001) Acute myeloid leukaemia cells secrete a soluble factor that inhibits T and NK cell proliferation but not cytolytic function—implications for the adoptive immunotherapy of leukaemia. Clin Exp Immunol 126:403CrossRefPubMedGoogle Scholar
  22. 22.
    Hirst WJ, Buggins A, Darling D, Gaken J, Farzaneh F, Mufti GJ (1997) Enhanced immune costimulatory activity of primary acute myeloid leukaemia blasts after retrovirus-mediated gene transfer of B7.1. Gene Ther 4:691CrossRefPubMedGoogle Scholar
  23. 23.
    Buggins AG, Lea N, Gaken J, Darling D, Farzaneh F, Mufti GJ, Hirst WJ (1999) Effect of costimulation and the microenvironment on antigen presentation by leukemic cells. Blood 94:3479PubMedGoogle Scholar
  24. 24.
    Chan L, Hardwick N, Darling D, Galea-Lauri J, Gaken J, Devereux S, Kemeny M, Mufti G, Farzaneh F (2005) IL-2/B7.1 (CD80) fusagene transduction of AML blasts by a self-inactivating lentiviral vector stimulates T cell responses in vitro: a strategy to generate whole cell vaccines for AML. Mol Ther 11:120CrossRefPubMedGoogle Scholar
  25. 25.
    Stripecke R, Cardoso AA, Pepper KA, Skelton DC, Yu XJ, Mascarenhas L, Weinberg KI, Nadler LM, Kohn DB (2000) Lentiviral vectors for efficient delivery of CD80 and granulocyte–macrophage- colony-stimulating factor in human acute lymphoblastic leukaemia and acute myeloid leukaemia cells to induce antileukemic immune responses. Blood 96:1317PubMedGoogle Scholar
  26. 26.
    Mutis T, Schrama E, Melief CJ, Goulmy E (1998) CD80-Transfected acute myeloid leukaemia cells induce primary allogeneic T-cell responses directed at patient specific minor histocompatibility antigens and leukaemia-associated antigens. Blood 92:1677PubMedGoogle Scholar
  27. 27.
    Koya RC, Kasahara N, Pullarkat V, Levine AM, Stripecke R (2002) Transduction of acute myeloid leukaemia cells with third generation self-inactivating lentiviral vectors expressing CD80 and GM-CSF: effects on proliferation, differentiation, and stimulation of allogeneic and autologous anti-leukaemia immune responses. Leukaemia 16:1645CrossRefGoogle Scholar
  28. 28.
    Notter M, Willinger T, Erben U, Thiel E (2001) Targeting of a B7-1 (CD80) immunoglobulin G fusion protein to acute myeloid leukaemia blasts increases their costimulatory activity for autologous remission T cells. Blood 97:3138CrossRefPubMedGoogle Scholar
  29. 29.
    Orleans-Lindsay JK, Deru A, Craig JI, Prentice HG, Lowdell MW (2003) In vitro co-stimulation with anti-CD28 synergizes with IL-12 in the generation of T cell immune responses to leukaemic cells; a strategy for ex-vivo generation of CTL for immunotherapy. Clin Exp Immunol 133:467CrossRefPubMedGoogle Scholar
  30. 30.
    Bruserud O, Ulvestad E (2000) Cytokine responsiveness of mitogen-activated T cells derived from acute leukaemia patients with chemotherapy-induced leukopenia. J Interferon Cytokine Res 20:947CrossRefPubMedGoogle Scholar
  31. 31.
    Dunussi-Joannopoulos K, Weinstein HJ, Nickerson PW, Strom TB, Burakoff SJ, Croop JM, Arceci RJ (1996) Irradiated B7-1 transduced primary acute myelogenous leukaemia (AML) cells can be used as therapeutic vaccines in murine AML. Blood 87:2938PubMedGoogle Scholar
  32. 32.
    Boyer MW, Vallera DA, Taylor PA, Gray GS, Katsanis E, Gorden K, Orchard PJ, Blazar BR (1997) The role of B7 costimulation by murine acute myeloid leukaemia in the generation and function of a CD8+ T-cell line with potent in vivo graft-versus-leukaemia properties. Blood 89:3477PubMedGoogle Scholar
  33. 33.
    Meloni G, Foa R, Vignetti M, Guarini A, Fenu S, Tosti S, Tos AG, Mandelli F (1994) Interleukin-2 may induce prolonged remissions in advanced acute myelogenous leukaemia. Blood 84:2158PubMedGoogle Scholar
  34. 34.
    Meloni G, Trisolini SM, Capria S, Torelli GF, Baldacci E, Torromeo C, Valesini G, Mandelli F (2002) How long can we give interleukin-2? Clinical and immunological evaluation of AML patients after 10 or more years of IL2 administration. Leukaemia 16:2016CrossRefGoogle Scholar
  35. 35.
    Hicks C, Cheung C, Lindeman R (2003) Restimulation of tumour-specific immunity in a patient with AML following injection with B7-1 positive autologous blasts. Leuk Res 27:1051CrossRefPubMedGoogle Scholar
  36. 36.
    Zhang WG, Liu SH, Cao XM, Cheng YX, Ma XR, Yang Y, Wang YL (2005) A phase-I clinical trial of active immunotherapy for acute leukaemia using inactivated autologous leukaemia cells mixed with IL-2, GM-CSF, and IL-6. Leuk Res 29:3CrossRefPubMedGoogle Scholar
  37. 37.
    Jedema I, van der Werff NM, Barge RM, Willemze R, Falkenburg JH (2004) New CFSE-based assay to determine susceptibility to lysis by cytotoxic T cells of leukemic precursor cells within a heterogeneous target cell population. Blood 103:2677CrossRefPubMedGoogle Scholar
  38. 38.
    Dermime S, Mavroudis D, Jiang YZ, Hensel N, Molldrem J, Barrett AJ (1997) Immune escape from a graft-versus-leukaemia effect may play a role in the relapse of myeloid leukemias following allogeneic bone marrow transplantation. Bone Marrow Transplant 19:989CrossRefPubMedGoogle Scholar
  39. 39.
    Lehmann C, Zeis M, Schmitz N, Uharek L (2000) Impaired binding of perforin on the surface of tumor cells is a cause of target cell resistance against cytotoxic effector cells. Blood 96:594PubMedGoogle Scholar
  40. 40.
    Ohnishi K, Yamanishi H, Naito K, Utsumi M, Yokomaku S, Hirabayashi N, Ohno R (1998) Reconstitution of peripheral blood lymphocyte subsets in the long-term disease-free survivors of patients with acute myeloblastic leukaemia. Leukaemia 12:52CrossRefGoogle Scholar
  41. 41.
    Buzyn A, Petit F, Ostankovitch M, Figueiredo S, Varet B, Guillet JG, Ameisen JC, Estaquier J (1999) Membrane-bound Fas (Apo-1/CD95) ligand on leukemic cells: a mechanism of tumor immune escape in leukaemia patients. Blood 94:3135PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Nicola Hardwick
    • 1
  • Lucas Chan
    • 1
  • Wendy Ingram
    • 1
  • Ghulam Mufti
    • 1
  • Farzin Farzaneh
    • 1
    Email author
  1. 1.Department of Haematological Medicine, King’s College LondonThe Rayne InstituteLondonUK

Personalised recommendations