Analytic and Dynamic Secretory Profile of Patient-Derived Cytokine-lnduced Killer Cells

Adoptive immunotherapy with cytokine induced killer (CIK) cells has shown antitumor activity against several kinds of cancer in preclinical models and clinical trials. CIK cells are a subset of ex vivo expanded T lymphocytes with T-NK phenotype and MHC-unrestricted antitumor activity. The literature provides scant information on cytokines, chemokines and growth factors secreted by CIK cells. Therefore, we investigated the secretory profile of CIK cells generated from tumor patients. The secretome analysis was performed at specific time points (d 1, d 14 and d 21) of CIK cell expansion. Mature CIK cells (d 21) produce a great variety of interleukins and secreted proteins that can be divided into three groups based on their secretion quantity: high (interleukin [IL]-13, regulated on activation normal T cell expressed and secreted [RANTES] chemokine, MIP-1α and 1β), medium (IL-1Ra, IL-5, IL-8, IL-10, IL-17, IP-10, INF-γ, vascular endothelial growth factor [VEGF] and granulocyte-macrophage colony-stimulating factor [GM-CSF]) and low (IL-1β, IL-4, IL-6, IL-7, IL-9, IL-12, IL-15, eotaxin, platelet-derived growth factor-bb, basic fibroblast growth factor, G-CSF and monocyte chemoattractant protein [MCP]-1). Moreover, comparing peripheral blood mononuclear cells (PBMCs) (d 1) and mature CIK cells (d 14 and 21) secretomes, we observed that IL-5, IL-10, IL-13, GM-CSF and VEGF were greatly upregulated, while IL-1β, IL-6, IL-8, IL-15, IL-17, eotaxin, MCP-1 and RANTES were downregulated. We also performed a gene expression profile analysis of patient-derived CIK cells, showing that mRNA for the different cytokines and secreted proteins was modulated during PBMC-to-CIK differentiation. We highlight previously unknown secretory properties and provide, for the first time, a comprehensive molecular characterization of CIK cells. Our findings provide a rationale to explore the functional implications and possible therapeutic modulation of CIK secretome.

[1]  E. Celis,et al.  Cancer immunotherapy: moving forward with peptide T cell vaccines. , 2017, Current opinion in immunology.

[2]  M. Colombo,et al.  Matricellular proteins tune myeloid‐derived suppressor cell recruitment and function in breast cancer , 2017, Journal of leukocyte biology.

[3]  K. Lim,et al.  Immunotherapy in gastrointestinal cancers. , 2017, Journal of gastrointestinal oncology.

[4]  Y. Matsumura,et al.  Therapeutic vaccination based on side population cells transduced by the granulocyte–macrophage colony-stimulating factor gene elicits potent antitumor immunity , 2017, Cancer Gene Therapy.

[5]  B. Li,et al.  Chemerin has a protective role in hepatocellular carcinoma by inhibiting the expression of IL-6 and GM-CSF and MDSC accumulation , 2017, Oncogene.

[6]  I. Zizzari,et al.  Interleukin-15 enhances cytokine induced killer (CIK) cytotoxic potential against epithelial cancer cell lines via an innate pathway. , 2016, Human immunology.

[7]  Burkhard Becher,et al.  GM-CSF: From Growth Factor to Central Mediator of Tissue Inflammation. , 2016, Immunity.

[8]  D. Ritchie,et al.  CIK immunotherapy in refractory hematologic malignancies. , 2016, Leukemia research.

[9]  Li Zhang,et al.  Systemic GM-CSF Recruits Effector T Cells into the Tumor Microenvironment in Localized Prostate Cancer , 2016, Cancer Immunology Research.

[10]  T. Itoh,et al.  Tumor‐Associated Macrophages Associate with Cerebrospinal Fluid Interleukin‐10 and Survival in Primary Central Nervous System Lymphoma (PCNSL) , 2016, Brain pathology.

[11]  Guoying Ni,et al.  Blocking IL-10 signalling at the time of immunization renders the tumour more accessible to T cell infiltration in mice. , 2016, Cellular immunology.

[12]  P. Piña-Sánchez,et al.  Mesenchymal stromal cells derived from cervical cancer tumors induce TGF-β1 expression and IL-10 expression and secretion in the cervical cancer cells, resulting in protection from cytotoxic T cell activity. , 2015, Cytokine.

[13]  J. Galić,et al.  Expression of proinflammatory cytokine interleukin-6 in tissue samples of human prostate obtained by needle biopsy. , 2015, Pathology, research and practice.

[14]  A. Goel,et al.  Functional genetic variability at promoters of pro‐(IL‐18) and anti‐(IL‐10) inflammatory affects their mRNA expression and survival in prostate carcinoma patients: Five year follow‐up study , 2015, The Prostate.

[15]  A. Głowacka,et al.  Inhibitors of Angiogenesis in Cancer Therapy - Synthesis and Biological Activity. , 2015, Current medicinal chemistry.

[16]  Jian Huang,et al.  Serum IL-10 Predicts Worse Outcome in Cancer Patients: A Meta-Analysis , 2015, PloS one.

[17]  Ruiqin Han,et al.  A novel anti-cancer agent Icaritin suppresses hepatocellular carcinoma initiation and malignant growth through the IL-6/Jak2/Stat3 pathway , 2015, Oncotarget.

[18]  E. Giannoni,et al.  Integrated gene and miRNA expression analysis of prostate cancer associated fibroblasts supports a prominent role for interleukin-6 in fibroblast activation , 2015 .

[19]  N. Lemoine,et al.  New role of Interleukin-10 in enhancing the antitumor efficacy of oncolytic vaccinia virus for treatment of pancreatic cancer , 2015, Oncoimmunology.

[20]  W. Paul,et al.  Innate Immune Function of TH2 Cells in vivo , 2015, Nature Immunology.

[21]  C. Coch,et al.  Cytokine-induced killer (CIK) cells in cancer immunotherapy: report of the international registry on CIK cells (IRCC) , 2015, Journal of Cancer Research and Clinical Oncology.

[22]  M. Wasik.  IL-13 as a novel growth factor in CTCL. , 2015, Blood.

[23]  S. Markovic,et al.  Adjuvant GM-CSF Improves Survival in High-risk Stage IIIC Melanoma: A Single-center Study , 2014, American journal of clinical oncology.

[24]  I. Schmidt-Wolf,et al.  An update on new adoptive immunotherapy strategies for solid tumors with cytokine-induced killer cells , 2014, Expert opinion on biological therapy.

[25]  C. Slingluff,et al.  Current status of granulocyte–macrophage colony-stimulating factor in the immunotherapy of melanoma , 2014, Journal of Immunotherapy for Cancer.

[26]  H. Abken,et al.  Arming cytokine-induced killer cells with chimeric antigen receptors: CD28 outperforms combined CD28-OX40 "super-stimulation". , 2013, Molecular therapy : the journal of the American Society of Gene Therapy.

[27]  D. Quail,et al.  Microenvironmental regulation of tumor progression and metastasis , 2013, Nature Medicine.

[28]  Huiping Wang,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 , 2013, Journal of immunotherapy.

[29]  M. Introna,et al.  Cytokine Induced Killer (CIK) cells for the treatment of haematological neoplasms. , 2013, Immunology letters.

[30]  Yantao Tian,et al.  Preclinical evaluation of herpes simplex virus armed with granulocyte-macrophage colony-stimulating factor in pancreatic carcinoma. , 2013, World journal of gastroenterology.

[31]  T. Venesio,et al.  Effective Activity of Cytokine-Induced Killer Cells against Autologous Metastatic Melanoma Including Cells with Stemness Features , 2013, Clinical Cancer Research.

[32]  Jingting Jiang,et al.  Cytokine-induced killer cells promote antitumor immunity , 2013, Journal of Translational Medicine.

[33]  W. Piacibello,et al.  Cytokine-induced killer (CIK) cells as feasible and effective adoptive immunotherapy for the treatment of solid tumors , 2012, Expert opinion on biological therapy.

[34]  X. Hao,et al.  Autologous cytokine-induced killer cell immunotherapy in lung cancer: a phase II clinical study , 2012, Cancer Immunology, Immunotherapy.

[35]  N. Cheung,et al.  Activation of peripheral-blood granulocytes is strongly correlated with patient outcome after immunotherapy with anti-GD2 monoclonal antibody and granulocyte-macrophage colony-stimulating factor. , 2012, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[36]  L. Moretta,et al.  Dual-functional capability of CD3+CD56+ CIK cells, a T-cell subset that acquires NK function and retains TCR-mediated specific cytotoxicity. , 2011, Blood.

[37]  D. Stocken,et al.  Elevated myeloid-derived suppressor cells in pancreatic, esophageal and gastric cancer are an independent prognostic factor and are associated with significant elevation of the Th2 cytokine interleukin-13 , 2011, Cancer Immunology, Immunotherapy.

[38]  R. Schreiber,et al.  Natural innate and adaptive immunity to cancer. , 2011, Annual review of immunology.

[39]  K. Hui,et al.  Cytokine-Induced NK-Like T Cells: From Bench to Bedside , 2010, Journal of biomedicine & biotechnology.

[40]  A. Contento,et al.  Immunotherapy with cytokine induced killer cells in solid and hematopoietic tumours: a pilot clinical trial , 2009, Hematological oncology.

[41]  Y. Shu,et al.  CD137 enhances cytotoxicity of CD3(+)CD56(+) cells and their capacities to induce CD4(+) Th1 responses. , 2009, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[42]  R. Bicknell,et al.  Anticancer strategies involving the vasculature , 2009, Nature Reviews Clinical Oncology.

[43]  M. Introna,et al.  Cytokine-induced killer cells are terminally differentiated activated CD8 cytotoxic T-EMRA lymphocytes. , 2009, Experimental hematology.

[44]  Gabriele Bergers,et al.  Modes of resistance to anti-angiogenic therapy , 2008, Nature Reviews Cancer.

[45]  M. Aglietta,et al.  Alloreactivity and anti-tumor activity segregate within two distinct subsets of cytokine-induced killer (CIK) cells: implications for their infusion across major HLA barriers. , 2008, International immunology.

[46]  R. Gambari,et al.  Evaluation of chemokine and cytokine profiles in osteoblast progenitors from umbilical cord blood stem cells by BIO‐PLEX technology , 2008, Cell biology international.

[47]  Hui Li,et al.  CD4+CD25+ Regulatory T Cells Decreased the Antitumor Activity of Cytokine-Induced Killer (CIK) Cells of Lung Cancer Patients , 2007, Journal of Clinical Immunology.

[48]  Robert S Negrin,et al.  Role of NKG2D signaling in the cytotoxicity of activated and expanded CD8+ T cells. , 2004, Blood.

[49]  Rafael A Irizarry,et al.  Exploration, normalization, and summaries of high density oligonucleotide array probe level data. , 2003, Biostatistics.

[50]  S. Shinkaruk,et al.  Vascular endothelial cell growth factor (VEGF), an emerging target for cancer chemotherapy. , 2003 .

[51]  A. Diefenbach,et al.  The role of the NKG2D immunoreceptor in immune cell activation and natural killing. , 2002, Immunity.

[52]  R. Negrin,et al.  Studies of ex Vivo Activated and Expanded CD8+ NK-T Cells in Humans and Mice , 2002, Journal of Clinical Immunology.

[53]  R. Negrin,et al.  Expansion of cytolytic CD8(+) natural killer T cells with limited capacity for graft-versus-host disease induction due to interferon gamma production. , 2001, Blood.

[54]  R. Armitage,et al.  ULBPs, novel MHC class I-related molecules, bind to CMV glycoprotein UL16 and stimulate NK cytotoxicity through the NKG2D receptor. , 2001, Immunity.

[55]  N. Shastri,et al.  Ligands for the murine NKG2D receptor: expression by tumor cells and activation of NK cells and macrophages , 2000, Nature Immunology.

[56]  D. Huhn,et al.  Phase I clinical study applying autologous immunological effector cells transfected with the interleukin-2 gene in patients with metastatic renal cancer, colorectal cancer and lymphoma , 1999, British Journal of Cancer.

[57]  T. Spies,et al.  Broad tumor-associated expression and recognition by tumor-derived γδ T cells of MICA and MICB , 1999 .

[58]  D D Donaldson,et al.  Interleukin-13: central mediator of allergic asthma , 1998 .

[59]  R. Negrin,et al.  A novel population of expanded human CD3+CD56+ cells derived from T cells with potent in vivo antitumor activity in mice with severe combined immunodeficiency. , 1994, Journal of immunology.

[60]  D. Huhn,et al.  Propagation of large numbers of T cells with natural killer cell markers , 1994, British journal of haematology.

[61]  P. Casellas,et al.  lnterleukin-13 is a new human lymphokine regulating inflammatory and immune responses , 1993, Nature.

[62]  I. Weissman,et al.  Use of a SCID mouse/human lymphoma model to evaluate cytokine-induced killer cells with potent antitumor cell activity , 1991, The Journal of experimental medicine.

[63]  A. Bertotti,et al.  Cytokine-induced killer cells eradicate bone and soft-tissue sarcomas. , 2014, Cancer research.

[64]  S. Fulda,et al.  The cytotoxic potential of interleukin-15-stimulated cytokine-induced killer cells against leukemia cells. , 2012, Cytotherapy.

[65]  R. Negrin,et al.  Role of NKG 2 D signaling in the cytotoxicity of activated and expanded CD 8 T cells , 2004 .

[66]  S. Bauer,et al.  Broad tumor-associated expression and recognition by tumor-derived gamma delta T cells of MICA and MICB. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[67]  M. Kaghad,et al.  Interleukin-13 is a new human lymphokine regulating inflammatory and immune responses. , 1993, Nature.

[68]  Wenqi Jiang,et al.  Oncotargets and Therapy Dovepress Increased Serum Levels of Interleukin-10 Predict Poor Prognosis in Extranodal Natural Killer/t-cell Lymphoma Patients Receiving Asparaginase-based Chemotherapy , 2022 .