Cytokine-Induced Killer Cells as an Adoptive Cellular Immunotherapy Strategy for Hepatocellular Carcinoma
Biochemistry and Molecular Biology
Volume 4, Issue 1, January 2019, Pages: 6-16
Received: Mar. 13, 2019;
Accepted: Apr. 13, 2019;
Published: May 17, 2019
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Nahla El-Sayed El-Ashmawy, Department of Biochemistry, Faculty of Pharmacy, Tanta University, Tanta, Egypt
Enas Arafa El-Zamarany, Department of Biochemistry, Faculty of Pharmacy, Tanta University, Tanta, Egypt
Hoda Abd El-Kader El-Bahrawy, Department of Clinical Pathology, Faculty of Medicine, Tanta University, Tanta, Egypt
Enas Abd El-Moneim Zahran, Department of Biochemistry, Faculty of Pharmacy, Tanta University, Tanta, Egypt
Background: Hepatocellular carcinoma (HCC) is the most common histologic type of primary liver cancer. HCC is the second highest mortality rate out of all major malignant carcinomas worldwide. Objectives: The aims of this study were to establish a rapid and easily handled culture method for sufficient expansion of viable and cytotoxic cytokine-induced killer (CIK) cells against HCC. Also, this study aimed to examine the morphologic, phenotypic, and functional characteristics of CIK cells. Method: Peripheral blood mononuclear cells (PBMCs) were cultured in a preliminary static culture to remove adherent cells. The suspended cells were cultured for 14 days with interferon-γ, human monoclonal anti-CD3 antibody and interleukin‑2. Aliquots of induced PBMCs were harvested weekly to assess informative morphologic and phenotypic features of CIK cells. Mature CIK cells were subjected to functional assays that included the production of TNFα and the cytotoxic effect on HCC cell line, HepG2. Findings: CIK cells could be successfully expanded from all samples with a significant increase in T cells, natural killer cells, and natural killer T cells. TNFα concentration in the culture supernatant was significantly increased. The cytotoxic effect of CIK cells on HepG2 cells was nearly 60% at 40:1, effector: target ratio. Regression analysis was used to predict the CIK: HepG2 ratio required to achieve complete cytotoxicity. Conclusion: This study provides a detailed and simple strategy for culturing effective CIK cells. Mature CIK cells showed a high functional capacity against HCC; which will support the further ongoing practice of immunotherapy integration into different current cancer treatment protocols.
Nahla El-Sayed El-Ashmawy,
Enas Arafa El-Zamarany,
Hoda Abd El-Kader El-Bahrawy,
Enas Abd El-Moneim Zahran,
Cytokine-Induced Killer Cells as an Adoptive Cellular Immunotherapy Strategy for Hepatocellular Carcinoma, Biochemistry and Molecular Biology.
Vol. 4, No. 1,
2019, pp. 6-16.
Ferlay J, Soerjomataram I, Dikshit R, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. International journal of cancer. 2015; 136 (5): E359-86. doi: 10.1002/ijc.29210.
van Ginneken V, de Vries E, Verheij E, et al. Potential Biomarkers for ‘Fatty Liver’(Hepatic Steatosis) and Hepatocellular Carcinoma (HCC) and an explanation of their pathogenesis. Gastroenterology and Liver Clinical and Medicals. 2017; 1 (1): 1-14.
Dhanasekaran R, Bandoh S, Roberts LR. Molecular pathogenesis of hepatocellular carcinoma and impact of therapeutic advances. F1000Research. 2016; 5 (879). doi: 10.12688/f1000research.6946.1.
Ghouri YA, Mian I, Rowe JH. Review of hepatocellular carcinoma: Epidemiology, etiology, and carcinogenesis. Journal of carcinogenesis. 2017; 16 (1). doi: 10.4103/jcar.JCar_9_16.
Agosti P, Sabbà C, Mazzocca A. Emerging metabolic risk factors in hepatocellular carcinoma and their influence on the liver microenvironment. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease. 2018;1864(2):607-617. doi: 10.1016/j.bbadis.2017.11.026.
Horst AK, Neumann K, Diehl L, et al. Modulation of liver tolerance by conventional and nonconventional antigen-presenting cells and regulatory immune cells. Cellular & molecular immunology. 2016;13 (3): 277-292. doi: 10.1038/cmi.2015.112.
Ma W, Chen X, Yuan Y. T-cell-associated immunotherapy: a promising strategy for the treatment of hepatocellular carcinoma. Immunotherapy. 2017; 9 (7): 523-525. doi: 10.2217/imt-2017-0053.
Sachdeva M, Chawla YK, Arora SK. Immunology of hepatocellular carcinoma. World Journal of Hepatology. 2015; 7 (17): 2080-2090. doi: 10.4254/wjh.v7.i17.2080.
Parker KH, Beury DW, Ostrand-Rosenberg S. Myeloid-Derived Suppressor Cells: Critical Cells Driving Immune Suppression in the Tumor Microenvironment. Advances in cancer research. 2015; 128: 95-139. doi: 10.1016/bs.acr.2015.04.002.
Orcutt S, A. Anaya D. Liver Resection and Surgical Strategies for Management of Primary Liver Cancer. Cancer Control. 2018; 25 (1). doi: 10.1177/1073274817744621
Rich NE, Yopp AC, Singal AG. Medical Management of Hepatocellular Carcinoma. Journal of oncology practice. 2017; 13 (6): 356-364. doi: 10.1200/jop.2017.022996.
Baradaran Noveiry B, Hirbod-Mobarakeh A, Khalili N, et al. Specific immunotherapy in hepatocellular cancer: A systematic review. Journal of gastroenterology and hepatology. 2017; 32 (2): 339-351. doi: 10.1111/jgh.13449.
Vu BT, Duong QT-N, Le PM, et al., editors. Culture and differentiation of cytokine-induced killer cells from umbilical cord blood-derived mononuclear cells. International Conference on the Development of Biomedical Engineering in Vietnam; 2017: Springer.
Lim O, Jung MY, Hwang YK, et al. Present and Future of Allogeneic Natural Killer Cell Therapy. Frontiers in Immunology. 2015; 6: 286. doi: 10.3389/fimmu.2015.00286.
Rosenberg SA, Restifo NP. Adoptive cell transfer as personalized immunotherapy for human cancer. Science. 2015; 348 (6230): 62-68. doi: 10.1126/science.aaa4967.
Introna M, Correnti F. Innovative Clinical Perspectives for CIK Cells in Cancer Patients. International Journal of Molecular Sciences. 2018; 19 (2): 358-371. doi: 10.3390/ijms19020358.
Mata-Molanes JJ, Sureda González M, Valenzuela Jiménez B, et al. Cancer Immunotherapy with Cytokine-Induced Killer Cells. Targeted Oncology. 2017; 12 (3): 289-299. doi: 10.1007/s11523-017-0489-2.
Introna M. CIK as therapeutic agents against tumors. Journal of Autoimmunity. 2017; 85: 32-44. doi: 10.1016/j.jaut.2017.06.008.
Cappuzzello E, Sommaggio R, Zanovello P, et al. Cytokines for the induction of antitumor effectors: The paradigm of Cytokine-Induced Killer (CIK) cells. Cytokine & Growth Factor Reviews. 2017; 36: 99-105. doi: 10.1016/j.cytogfr.2017.06.003.
Guo Y, Han W. Cytokine-induced killer (CIK) cells: from basic research to clinical translation. Chinese Journal of Cancer. 2015; 34 (3). doi: 10.1186/s40880-015-0002-1.
Meng Y, Yu Z, Wu Y, et al. Cell-based immunotherapy with cytokine-induced killer (CIK) cells: From preparation and testing to clinical application. Human Vaccines & Immunotherapeutics. 2017; 13 (6): 1379-1387. doi: 10.1080/21645515.2017.1285987.
Janetzki S. Sample Preparation. In: Janetzki S, editor. Elispot for Rookies (and Experts Too). Cham: Springer International Publishing; 2016. p. 25-41.
Riedhammer C, Halbritter D, Weissert R. Peripheral Blood Mononuclear Cells: Isolation, Freezing, Thawing, and Culture. Methods in molecular biology. 2016; 1304:53-61. doi: 10.1007/7651_2014_99.
Kerbel RS, Blakeslee D. Rapid adsorption of a foetal calf serum component by mammalian cells in culture. A potential source of artifacts in studies of antisera to cell-specific antigens. Immunology. 1976; 31 (6): 881-891.
Reddy BP, Reddy BP, Rayulu DJ. Effects of fetal bovine serum concentration on the growth and survival of BHK 21 cell lines. International Journal of Applied Biology and Pharmaceutical Technology. 2016; 7 (2): 122-127.
Hontscha C, Borck Y, Zhou H, et al. Clinical trials on CIK cells: first report of the international registry on CIK cells (IRCC). Journal of Cancer Research and Clinical Oncology. 2011; 137 (2): 305-310. doi: 10.1007/s00432-010-0887-7.
Schmidt TL, Negrin RS, Contag CH. A killer choice for cancer immunotherapy. Immunologic Research. 2014; 58 (2): 300-306. doi: 10.1007/s12026-014-8507-2.
Mitra S, Ring AM, Amarnath S, et al. Interleukin-2 activity can be fine tuned with engineered receptor signaling clamps. Immunity. 2015; 42 (5): 826-838. doi: 10.1016/j.immuni.2015.04.018.
Iudicone P, Fioravanti D, Cicchetti E, et al. Interleukin-15 enhances cytokine induced killer (CIK) cytotoxic potential against epithelial cancer cell lines via an innate pathway. Human Immunology. 2016; 77 (12): 1239-1247. doi: 10.1016/j.humimm.2016.09.003.
Guo Q, Zhu D, Bu X, et al. Efficient killing of radioresistant breast cancer cells by cytokine-induced killer cells. Tumour Biology. 2017; 39 (3). doi: 10.1177/1010428317695961.
Niam M, Linn YC, Fook Chong S, et al. Clinical scale expansion of cytokine-induced killer cells is feasible from healthy donors and patients with acute and chronic myeloid leukemia at various stages of therapy. Experimental hematology. 2011; 39 (9): 897-903. doi: 10.1016/j.exphem.2011.06.005.
Liu H, Li J, Wang F, et al. Comparative study of different procedures for the separation of peripheral blood mononuclear cells in cytokine-induced killer cell immunotherapy for hepatocarcinoma. Tumour biology. 2015; 36 (4): 2299-3307. doi: 10.1007/s13277-014-2837-5.
Chan W-C, Linn Y-C. A comparison between cytokine-and bead-stimulated polyclonal T cells: the superiority of each and their possible complementary role. Cytotechnology. 2016; 68 (4): 735-748. doi: 10.1007/s10616-014-9825-x.
Zhang Z, Zhao X, Zhang T, et al. Phenotypic characterization and anti-tumor effects of cytokine-induced killer cells derived from cord blood. Cytotherapy. 2015;17 (1): 86-97. doi: 10.1016/j.jcyt.2014.09.006.
Zhang Q, Wang L, Luo C, et al. Phenotypic and functional characterization of cytokine-induced killer cells derived from preterm and term infant cord blood. Oncology Reports. 2014; 32 (5): 2244-2252. doi: 10.3892/or.2014.
Durrieu L, Lemieux W, Dieng MM, et al. Implication of different effector mechanisms by cord blood-derived and peripheral blood-derived cytokine-induced killer cells to kill precursor B acute lymphoblastic leukemia cell lines. Cytotherapy. 2014; 16 (6): 845-56. doi: 10.1016/j.jcyt.2013.12.010.
Li H, Huang L, Liu L, et al. Selective effect of cytokine-induced killer cells on survival of patients with early-stage melanoma. Cancer immunology, immunotherapy. 2017;66(3):299-308. doi: 10.1007/s00262-016-1939-x.
Wei C, Wang W, Pang W, et al. The CIK cells stimulated with combination of IL-2 and IL-15 provide an improved cytotoxic capacity against human lung adenocarcinoma. Tumour biology. 2014; 35 (3): 1997-2007. doi: 10.1007/s13277-013-1265-2.
Helms MW, Prescher JA, Cao Y-A, et al. IL-12 enhances efficacy and shortens enrichment time in cytokine-induced killer cell immunotherapy. Cancer immunology, immunotherapy. 2010; 59 (9): 1325-1334. doi: 10.1007/s00262-010-0860-y.
Lin G, Wang J, Lao X, et al. Interleukin-6 inhibits regulatory T cells and improves the proliferation and cytotoxic activity of cytokine-induced killer cells. Journal of immunotherapy. 2012; 35 (4): 337-343. doi: 10.1097/CJI.0b013e318255ada3.
Tao Q, Chen T, Tao L, et al. IL-15 improves the cytotoxicity of cytokine-induced killer cells against leukemia cells by upregulating CD3+CD56+ cells and downregulating regulatory T cells as well as IL-35. Journal of immunotherapy. 2013; 36 (9): 462-467. doi: 10.1097/cji.0000000000000001.
Rajbhandary S, Zhao M-F, Zhao N, et al. Multiple cytotoxic factors involved in IL-21 enhanced antitumor function of CIK cells signaled through STAT-3 and STAT5b pathways. Asian Pacific Journal of Cancer Prevention. 2013; 14 (10): 5825-5831.
Ingersoll SB, Srivastava M, Ali G, et al. Cytokine-induced killer cells from ovarian cancer patients expanded ex vivo in the presence of IL-7 improve survival in a xenograft mouse model of ovarian cancer. Gynecologic Oncology. 2014; 133: 120-121. doi: 10.1016/j.ygyno.2014.03.316.
Mesiano G, Todorovic M, Gammaitoni L, et al. Cytokine-induced killer (CIK) cells as feasible and effective adoptive immunotherapy for the treatment of solid tumors. Expert opinion on biological therapy. 2012; 12 (6): 673-84. doi: 10.1517/14712598.2012.675323.
Li GX, Zhao SS, Zhang XG, et al. Comparison of the proliferation, cytotoxic activity and cytokine secretion function of cascade primed immune cells and cytokine-induced killer cells in vitro. Molecular Medicine Reports. 2015; 12 (2): 2629-2635. doi: 10.3892/mmr.2015.3765.
Bonanno G, Iudicone P, Mariotti A, et al. Thymoglobulin, interferon-γ and interleukin-2 efficiently expand cytokine-induced killer (CIK) cells in clinical-grade cultures. Journal of Translational Medicine. 2010; 8 (1): 129. doi: 10.1186/1479-5876-8-129.
Luo H, Gong L, Zhu B, et al. Therapeutic outcomes of autologous CIK cells as a maintenance therapy in the treatment of lung cancer patients: A retrospective study. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie. 2016; 84: 987-993. doi: 10.1016/j.biopha.2016.10.022.
Dembic Z. Chapter 8 - Cytokines Important for Growth and/or Development of Cells of the Immune System. In: Dembic Z, editor. The Cytokines of the Immune System. Amsterdam: Academic Press; 2015. p. 263-281.
Guo ZHI, Liu HAO, He X-P, et al. A clinical study of cytokine-induced killer cells for the treatment of refractory lymphoma. Oncology Letters. 2011; 2 (3): 531-536. doi: 10.3892/ol.2011.269.
Linn YC, Niam M, Chu S, et al. The anti-tumour activity of allogeneic cytokine-induced killer cells in patients who relapse after allogeneic transplant for haematological malignancies. Bone marrow transplantation. 2012; 47 (7): 957-966. doi: 10.1038/bmt.2011.202.
Kornacker M, Moldenhauer G, Herbst M, et al. Cytokine-induced killer cells against autologous CLL: direct cytotoxic effects and induction of immune accessory molecules by interferon-gamma. International journal of cancer. 2006; 119 (6): 1377-82. doi: 10.1002/ijc.21994.
Youlin K, Jian K, Siming L, et al. Potent anti-prostate cancer immune response induced by dendritic cells transduced with recombinant adenoviruses encoding 4-1BBL combined with cytokine-induced killer cells. Immunotherapy. 2015; 7 (1): 13-20. doi: 10.2217/imt.14.92.
Sangiolo D, Mesiano G, Gammaitoni L, et al. Cytokine-induced killer cells eradicate bone and soft-tissue sarcomas. Cancer Research. 2014; 74 (1): 119-129. doi: 10.1158/0008-5472.CAN-13-1559.
Gammaitoni L, Giraudo L, Leuci V, et al. Effective Activity of Cytokine-Induced Killer Cells against Autologous Metastatic Melanoma Including Cells with Stemness Features. Clinical Cancer Research. 2013; 19 (16): 4347-4358. doi: 10.1158/1078-0432.CCR-13-0061 %J Clinical Cancer Research.
Jin J, Joo KM, Lee SJ, et al. Synergistic therapeutic effects of cytokine-induced killer cells and temozolomide against glioblastoma. Oncology Reports. 2011; 25 (1): 33-39. doi: 10.3892/or_00001038.
Wang J, He M, Shi W, et al. Inducible costimulator (ICOS) enhances the cytolytic activity of cytokine-induced killer cells against gallbladder cancer in vitro and in vivo. Cancer investigation. 2009 Mar; 27 (3): 244-250. doi: 10.1080/07357900802239124.