The Effect of Pretreatment with Chemotherapeutic Drugs on the Susceptibility to Lymphokine Activated CD8+ T Lymphocyte-Mediated Cytotoxicity in CMK Leukemia Cells
Keywords:Leukemia, cytotoxic T lymphocytes, chemotherapy, apoptosis
Objectives: Certain tumor cells pretreated with chemotherapeutic drugs become more susceptible to death by apoptosis induced by killer cells of the immune system. We examined the CD8+ cytotoxic T lymphocyte (CTL)-mediated cytotoxicity in myeloid leukemia cell lines pretreated with chemotherapeutic drugs.
Methods: Peripheral blood mononuclear cells were expanded in vitro in the presence of phytohemagglutinin-P, interleukins-2 and -15. CD3+ CD8+ cells representing the CTLs were isolated using magnetic immunoselection and used in immune cytotoxicity experiments against K562 and CMK leukemia cells, pretreated with two different concentrations of cytarabine and etoposide.
Results: In CMK cells pretreated with etoposide at 2 M and 20 M concentrations, the mean cell-mediated immune cytotoxicity rose to 21.4 ± 12.9% (p=0.09) and 23.4 ± 12.6% (p=0.046), respectively, when compared to the control value of 6.6 ± 3.8%. In CMK cells pretreated with cytarabine at 1 M and 10M concentrations, the mean immune cytotoxicity rose to 14.3 ± 11.2% and 22.6 ± 15.2%, respectively, compared to the control value of 8.7 ± 6.3%, although these results did not reach statistical significance. However, a similar increase in CTL-mediated immune cytotoxicity was not observed against drug-treated K562 cells.
Conclusion: This study suggests that pretreatment with chemotherapeutic drugs can render CMK leukemia cells more susceptible to immune attack by activated CTLs. Further studies are needed to explore this phenomenon, to establish an immune-enhancing effect of pretreatment with chemotherapy in the treatment of leukemia.
Kolb HJ, et al. Graft-versus-leukemia reactions in allogeneic chimeras. Blood 2004; 103(3): 767-76. http://dx.doi.org/10.1182/blood-2003-02-0342
Shah NN, Dave H, Wayne AS. Immunotherapy for pediatric leukemia. Front Oncol 2013; 3: 166. http://dx.doi.org/10.3389/fonc.2013.00166
Horowitz MM, et al. Graft-versus-leukemia reactions after bone marrow transplantation. Blood 1990; 75(3): 555-62.
Drobyski WR, et al. Molecular remission occurring after donor leukocyte infusions for the treatment of relapsed chronic myelogenous leukemia after allogeneic bone marrow transplantation. Bone Marrow Transplant 1992; 10(3): 301-4.
Chang YJ, Huang XJ. Donor lymphocyte infusions for relapse after allogeneic transplantation: when, if and for whom? Blood Rev 2013; 27(1): 55-62. http://dx.doi.org/10.1016/j.blre.2012.11.002
Cheung NV, Rooney CM. Principles of immune and cellular therapy in Principles and Practice of Pediatric Oncology, P.A. Pizzo and D.G. Poplack, Editors. 2002, Lippincott Williams and Wilkins: Philadelphia. p. 381-408.
Takei F. LAK cell therapy of AML: not to be lost in translation. Exp Hematol 2011; 39(11): 1045-6. http://dx.doi.org/10.1016/j.exphem.2011.08.005
Russell JH, Ley TJ. Lymphocyte-mediated cytotoxicity. Annu Rev Immunol 2002; 20: 323-70. http://dx.doi.org/10.1146/annurev.immunol.20.100201.131730
Smyth MJ, et al. Nature's TRAIL-on a path to cancer immunotherapy. Immunity 2003; 18(1): 1-6. http://dx.doi.org/10.1016/S1074-7613(02)00502-2
Stuckey DW, Shah K. TRAIL on trial: preclinical advances in cancer therapy. Trends Mol Med 2013; 19(11): 685-94. http://dx.doi.org/10.1016/j.molmed.2013.08.007
Kelley SK, Ashkenazi A. Targeting death receptors in cancer with Apo2L/TRAIL. Curr Opin Pharmacol 2004; 4(4): 333-9. http://dx.doi.org/10.1016/j.coph.2004.02.006
Srivastava RK. TRAIL/Apo-2L: mechanisms and clinical applications in cancer. Neoplasia 2001; 3(6): 535-46. http://dx.doi.org/10.1038/sj.neo.7900203
Matsuzaki I, et al. Cisplatin induces fas expression in esophageal cancer cell lines and enhanced cytotoxicity in combination with LAK cells. Oncology 2000; 59(4): 336-43. http://dx.doi.org/10.1159/000012192
Wang MJ, et al. [Chemotherapeutic drugs enhanced rsTRAIL tumoricidal activity]. Zhongguo Yi Xue Ke Xue Yuan Xue Bao 2004; 26(5): 524-8.
Beranova L, et al. The plant alkaloid and anti-leukemia drug homoharringtonine sensitizes resistant human colorectal carcinoma cells to TRAIL-induced apoptosis via multiple mechanisms. Apoptosis 2013; 18(6): 739-50. http://dx.doi.org/10.1007/s10495-013-0823-9
Iwase M, et al. Enhanced susceptibility of oral squamous cell carcinoma cell lines to FAS-mediated apoptosis by cisplatin and 5-fluorouracil. Int J Cancer 2003; 106(4): 619-25. http://dx.doi.org/10.1002/ijc.11239
Mishima K, Nariai Y, Yoshimura Y. Carboplatin induces Fas (APO-1/CD95)-dependent apoptosis of human tongue carcinoma cells: sensitization for apoptosis by upregulation of FADD expression. Int J Cancer 2003; 105(5): 593-600. http://dx.doi.org/10.1002/ijc.11133
Mizutani H, et al. Mechanism of apoptosis induced by doxorubicin through the generation of hydrogen peroxide. Life Sci 2005; 76(13): 1439-53. http://dx.doi.org/10.1016/j.lfs.2004.05.040
Mizutani Y, et al. Enhanced susceptibility of cis-diamminedichloroplatinum-treated K562 cells to lysis by peripheral blood lymphocytes and lymphokine activated killer cells. Cancer 1993; 71(4): 1313-21. http://dx.doi.org/10.1002/1097-0142(19930215)71:4<1313::AID-CNCR2820710424>3.0.CO;2-
Mizutani Y, et al. Doxorubicin sensitizes human bladder carcinoma cells to Fas-mediated cytotoxicity. Cancer 1997; 79(6): 1180-9. http://dx.doi.org/10.1002/(SICI)1097-0142(19970315)79:6<1180::AID-CNCR17>3.0.CO;2-W
Mizutani Y, Yoshida O, Bonavida B. Sensitization of human bladder cancer cells to Fas-mediated cytotoxicity by cis-diamminedichloroplatinum (II). J Urol 1998; 160(2): 561-70. http://dx.doi.org/10.1016/S0022-5347(01)62959-8
Posovszky C, et al. Chemotherapeutic drugs sensitize pre-B ALL cells for CD95- and cytotoxic T-lymphocyte-mediated apoptosis. Leukemia 1999; 13(3): 400-9. http://dx.doi.org/10.1038/sj.leu.2401327
Uslu R, et al. Chemosensitization of human prostate carcinoma cell lines to anti-fas-mediated cytotoxicity and apoptosis. Clin Cancer Res 1997; 3(6): 963-72.
Uslu R, Jewett A, Bonavida B. Sensitization of human ovarian tumor cells by subtoxic CDDP to anti-fas antibody-mediated cytotoxicity and apoptosis. Gynecol Oncol 1996; 62(2): 282-91. http://dx.doi.org/10.1006/gyno.1996.0228
Wu XX, et al. Doxorubicin enhances TRAIL-induced apoptosis in prostate cancer. Int J Oncol 2002; 20(5): 949-54. http://dx.doi.org/10.1002/ijc.10948
Wu XX, et al. Enhancement of TRAIL/Apo2L-mediated apoptosis by adriamycin through inducing DR4 and DR5 in renal cell carcinoma cells. Int J Cancer 2003; 104(4): 409-17.
Wu XX, et al. Enhancement of Fas-mediated apoptosis in renal cell carcinoma cells by adriamycin. Cancer Res 2000; 60(11): 2912-8.
Yamamoto T, et al. Enhanced apoptosis of squamous cell carcinoma cells by interleukin-2-activated cytotoxic lymphocytes combined with radiation and anticancer drugs. Eur J Cancer 2000; 36(15): 2007-17. http://dx.doi.org/10.1016/S0959-8049(00)00244-6
Yamaue H, et al. Cisplatin treatment renders tumor cells more susceptible to attack by lymphokine-activated killer cells. J Clin Lab Immunol 1991; 35(4): 165-70.
Johnston JB, et al. Role of the TRAIL/APO2-L death receptors in chlorambucil- and fludarabine-induced apoptosis in chronic lymphocytic leukemia. Oncogene 2003; 22(51): 8356-69. http://dx.doi.org/10.1038/sj.onc.1207004
Jones DT, et al. Cytotoxic drugs enhance the ex vivo sensitivity of malignant cells from a subset of acute myeloid leukaemia patients to apoptosis induction by tumour necrosis factor receptor-related apoptosis-inducing ligand. Br J Haematol 2003; 121(5): 713-20. http://dx.doi.org/10.1046/j.1365-2141.2003.04340.x
Ozdemir O, Savasan S. Combinational IL-2/IL-15 induction does not further enhance IL-15-induced lymphokine-activated killer cell cytotoxicity against human leukemia/lymphoma cells. Clin Immunol 2005; 115(3): 240-9. http://dx.doi.org/10.1016/j.clim.2005.01.008
Savasan S, et al. Evaluation of cytotoxicity by flow cytometric drug sensitivity assay in childhood T-cell acute lymphoblastic leukemia. Leuk Lymphoma 2005; 46(6): 833-41. http://dx.doi.org/10.1080/10428190500079951
Lozzio BB, Lozzio CB. Properties of the K562 cell line derived from a patient with chronic myeloid leukemia. Int J Cancer 1977; 19(1): 136. http://dx.doi.org/10.1002/ijc.2910190119
Sato T, et al. Establishment of a human leukaemic cell line (CMK) with megakaryocytic characteristics from a Down's syndrome patient with acute megakaryoblastic leukaemia. Br J Haematol 1989; 72(2): 184-90. http://dx.doi.org/10.1111/j.1365-2141.1989.tb07681.x
Ozdemir O, Ravindranath Y, Savasan S, Cell-mediated cytotoxicity evaluation using monoclonal antibody staining for target or effector cells with annexinV/propidium iodide colabeling by fluorosphere-adjusted counts on three-color flow cytometry. Cytometry A 2003; 56(1): 53-60. http://dx.doi.org/10.1002/cyto.a.10081
Ruggeri L, et al. Natural killer cell alloreactivity in haploidentical hematopoietic stem cell transplantation. Int J Hematol 2005; 81(1): 13-7. http://dx.doi.org/10.1532/IJH97.04172
Locatelli F, et al. Cellular and molecular basis of haploidentical hematopoietic stem cell transplantation in the successful treatment of high-risk leukemias: role of alloreactive NK cells. Front Immunol 2013; 4: 15. http://dx.doi.org/10.3389/fimmu.2013.00015
Alves NL, et al. IL-15 induces antigen-independent expansion and differentiation of human naive CD8+ T cells in vitro. Blood 2003; 102(7): 2541-6. http://dx.doi.org/10.1182/blood-2003-01-0183
Weng NP, et al. IL-15 is a growth factor and an activator of CD8 memory T cells. Ann N Y Acad Sci 2002; 975: 46-56. http://dx.doi.org/10.1111/j.1749-6632.2002.tb05940.x
Sone, S, et al. Heterogeneity of human lymphokine (IL-2)-activated killer (LAK) precursors and regulation of their LAK induction by blood monocytes. Int J Cancer 1988; 42(3): 428-34. http://dx.doi.org/10.1002/ijc.2910420320
Taub JW, et al. Expression of chromosome 21-localized genes in acute myeloid leukemia: differences between Down syndrome and non-Down syndrome blast cells and relationship to in vitro sensitivity to cytosine arabinoside and daunorubicin. Blood 1999; 94(4): 1393-400.
Sreenivasan Y, Sarkar A, Manna SK. Mechanism of cytosine arabinoside-mediated apoptosis: role of Rel A (p65) dephosphorylation. Oncogene 2003; 22(28): 4356-69. http://dx.doi.org/10.1038/sj.onc.1206486
Dimery IW, et al. Variation amongst K562 cell cultures. Exp Hematol 1983; 11(7): 601-10.