{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T09:34:34Z","timestamp":1762508074105,"version":"build-2065373602"},"reference-count":29,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2020,4,27]],"date-time":"2020-04-27T00:00:00Z","timestamp":1587945600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Cytokine-induced killer (CIK) cells are heterogeneous, major histocompatibility complex (MHC)-unrestricted T lymphocytes that have acquired the expression of several natural killer (NK) cell surface markers following the addition of interferon gamma (IFN-\u03b3), OKT3 and interleukin-2 (IL-2). Treatment with CIK cells demonstrates a practical approach in cancer immunotherapy with limited, if any, graft versus host disease (GvHD) toxicity. CIK cells have been proposed and tested in many clinical trials in cancer patients by autologous, allogeneic or haploidentical administration. The possibility of combining them with specific monoclonal antibodies nivolumab and ipilimumab will further expand the possibility of their clinical utilization. Initially, phenotypic analysis was performed to explore CD3, CD4, CD56, PD-1 and CTLA-4 expression on CIK cells and PD-L1\/PD-L2 expression on tumor cells. We further treated CIK cells with nivolumab and ipilimumab and measured the cytotoxicity of CIK cells cocultured to renal carcinoma cell lines, A-498 and Caki-2. We observed a significant decrease in viability of renal cell lines after treating with CIK cells (p &lt; 0.0001) in comparison to untreated renal cell lines and anti-PD-1 or anti-CTLA-4 treatment had no remarkable effect on the viability of tumor cells. Using CCK-8, Precision Count Beads\u2122 and Cell Trace\u2122 violet proliferation assays, we proved significant increased proliferation of CIK cells in the presence of a combination of anti-PD-1 and anti-CTLA-4 antibodies compared to untreated CIK cells. The IFN-\u03b3 secretion increased significantly in the presence of A-498 and combinatorial blockade of PD-1 and CTLA-4 compared to nivolumab or ipilimumab monotreatment (p &lt; 0.001). In conclusion, a combination of immune checkpoint inhibition with CIK cells augments cytotoxicity of CIK cells against renal cancer cells.<\/jats:p>","DOI":"10.3390\/ijms21093078","type":"journal-article","created":{"date-parts":[[2020,4,28]],"date-time":"2020-04-28T05:05:32Z","timestamp":1588050332000},"page":"3078","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Increase in Efficacy of Checkpoint Inhibition by Cytokine-Induced-Killer Cells as a Combination Immunotherapy for Renal Cancer"],"prefix":"10.3390","volume":"21","author":[{"given":"Mojgan Naghizadeh","family":"Dehno","sequence":"first","affiliation":[{"name":"Department of Integrated Oncology, CIO Bonn, University Hospital Bonn, Venusberg-Campus 1, D 53127 Bonn, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0032-4940","authenticated-orcid":false,"given":"Yutao","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Integrated Oncology, CIO Bonn, University Hospital Bonn, Venusberg-Campus 1, D 53127 Bonn, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7521-9662","authenticated-orcid":false,"given":"Hans","family":"Weiher","sequence":"additional","affiliation":[{"name":"Department of Applied Natural Sciences, Bonn-Rhein-Sieg University of Applied Sciences, D-53359 Rheinbach, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ingo G.H.","family":"Schmidt-Wolf","sequence":"additional","affiliation":[{"name":"Department of Integrated Oncology, CIO Bonn, University Hospital Bonn, Venusberg-Campus 1, D 53127 Bonn, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,4,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1111\/j.1365-2141.1994.tb08297.x","article-title":"Propagation of large numbers of T cells with natural killer cell markers","volume":"87","author":"Lefterova","year":"1994","journal-title":"Br. J. Haematol."},{"key":"ref_2","first-page":"1673","article-title":"Phenotypic characterization and identification of effector cells involved in tumor cell recognition of cytokine-induced killer cells","volume":"21","author":"Lefterova","year":"1993","journal-title":"Exp. Hematol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1115","DOI":"10.1158\/2326-6066.CIR-15-0190","article-title":"T-cell-based immunotherapy: Adoptive cell transfer and checkpoint inhibition","volume":"3","author":"Houot","year":"2015","journal-title":"Cancer Immunol. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1186\/1479-5876-11-83","article-title":"Cytokine-induced killer cells promote antitumor immunity","volume":"11","author":"Jiang","year":"2013","journal-title":"J. Transl. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1186\/s40880-015-0002-1","article-title":"Cytokine-induced killer (CIK) cells: From basic research to clinical translation","volume":"34","author":"Guo","year":"2015","journal-title":"Chin. J. Cancer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1084\/jem.174.1.139","article-title":"Use of SCID mouse\/human lymphoma model to evaluate cytokine-induced killer cells with potent antitumor cell activity","volume":"174","author":"Negrin","year":"1991","journal-title":"J. Exp. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"14632","DOI":"10.3390\/ijms150814632","article-title":"Adoptive Immunotherapy Strategies with Cytokine-Induced Killer (CIK) Cells in the Treatment of Hematological Malignancies","volume":"15","author":"Schmeel","year":"2014","journal-title":"Int. J. Mol. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.imlet.2013.09.017","article-title":"Cytokine Induced Killer (CIK) cells for treatment of haematological neoplasms","volume":"155","author":"Introna","year":"2013","journal-title":"Immunol. Lett."},{"key":"ref_9","first-page":"Doc07","article-title":"Cytokine-induced Killer Cells Are Type II Natural Killer T Cells","volume":"5","author":"Guetgemann","year":"2007","journal-title":"GMS Ger. Med. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1007\/s00432-010-0887-7","article-title":"Clinical trials on CIK cells: First report of the international registry on CIK cells (IRCC)","volume":"137","author":"Hontscha","year":"2011","journal-title":"J. Cancer Res. Clin. Oncol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1212","DOI":"10.1038\/ni.2762","article-title":"A rheostat for immune responses: The unique properties of PD-1 and their advantages for clinical application","volume":"14","author":"Okazaki","year":"2013","journal-title":"Nat. Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1146\/annurev.immunol.26.021607.090331","article-title":"PD-1 and its ligands in tolerance and immunity","volume":"26","author":"Keir","year":"2008","journal-title":"Annu. Rev. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"847","DOI":"10.1517\/14712598.2013.770836","article-title":"Anti-programmed death-1 and anti-programmed death-ligand 1 antibodies in cancer therapy","volume":"13","author":"Hamid","year":"2013","journal-title":"Expert Opin. Biol. Ther."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2263","DOI":"10.1126\/science.282.5397.2263","article-title":"Molecular basis of T cell inactivation by CTLA-4","volume":"282","author":"Lee","year":"1998","journal-title":"Science"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1126\/science.1202947","article-title":"Trans-endocytosis of CD80 and CD86: A molecular basis for the cell-extrinsic function of CTLA-4","volume":"332","author":"Qureshi","year":"2011","journal-title":"Science"},{"key":"ref_16","first-page":"13","article-title":"Renal Cancer Tissue after Nivolumab\/Ipilimumab Combination Therapy for Metastatic Renal Cell Carcinoma","volume":"66","author":"Okuno","year":"2020","journal-title":"Hinyokika Kiyo"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/j.eururo.2019.05.022","article-title":"Updated European Association of Urology Guidelines on Renal Cell Carcinoma: Immune Checkpoint Inhibition Is the New Backbone in First-line Treatment of Metastatic Clear-cell Renal Cell Carcinoma","volume":"76","author":"Albiges","year":"2019","journal-title":"Eur. Urol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.1056\/NEJMoa1712126","article-title":"Nivolumab plus Ipilimumab versus Sunitinib in Advanced Renal-Cell Carcinoma","volume":"378","author":"Motzer","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1016\/S1470-2045(18)30778-2","article-title":"Patient-reported outcomes of patients with advanced renal cell carcinoma treated with nivolumab plus ipilimumab versus sunitinib (CheckMate 214): A randomised, phase 3 trial","volume":"20","author":"Cella","year":"2019","journal-title":"Lancet Oncol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1016\/S1470-2045(15)70076-8","article-title":"Nivolumab versus chemotherapy in patients with advanced melanoma who progressed after anti-CTLA-4 treatment (CheckMate 037): A randomised, controlled, open-label, phase 3 trial","volume":"16","author":"Weber","year":"2015","journal-title":"Lancet Oncol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1056\/NEJMoa1504627","article-title":"Nivolumab versus docetaxel in advanced squamous-cell non\u2013small-cell lung cancer","volume":"373","author":"Brahmer","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1627","DOI":"10.1056\/NEJMoa1507643","article-title":"Nivolumab versus docetaxel in advanced nonsquamous non\u2013small-cell lung cancer","volume":"373","author":"Borghaei","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1803","DOI":"10.1056\/NEJMoa1510665","article-title":"Nivolumab versus everolimus in advanced renal-cell carcinoma","volume":"373","author":"Motzer","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.oraloncology.2018.04.008","article-title":"Nivolumab vs Investigator\u2019s Choice in Recurrent or Metastatic Squamous Cell Carcinoma of the Head and Neck: 2-year Long-Term Survival Update of CheckMate 141 with Analyses by Tumor PD-L1 Expression","volume":"81","author":"Ferris","year":"2018","journal-title":"Oral Oncol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1016\/j.canlet.2015.03.043","article-title":"Cytokine induced killer cell-based immunotherapies in patients with different stages of renal cell carcinoma","volume":"362","author":"Zhao","year":"2015","journal-title":"Cancer Lett."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1583","DOI":"10.1111\/cas.13560","article-title":"High IDO-1 expression in tumor endothelial cells is associated with response to immunotherapy in metastatic renal cell carcinoma","volume":"109","author":"Seeber","year":"2018","journal-title":"Cancer Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1186\/s12943-016-0565-8","article-title":"Choosing the right cell line for renal cell cancer research","volume":"15","author":"Brodaczewska","year":"2016","journal-title":"Mol. Cancer."},{"key":"ref_28","first-page":"S21","article-title":"Combining differential expression, chromosomal and pathway analyses for the molecular characterization of renal cell carcinoma","volume":"1","author":"Furge","year":"2007","journal-title":"Can. Urol. Assoc. J."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1038\/nrc3984","article-title":"MYC: Connecting selective transcriptional control to global RNA production","volume":"15","author":"Kress","year":"2015","journal-title":"Nat. Rev. Cancer"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/9\/3078\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T13:23:55Z","timestamp":1760361835000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/9\/3078"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,4,27]]},"references-count":29,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2020,5]]}},"alternative-id":["ijms21093078"],"URL":"https:\/\/doi.org\/10.3390\/ijms21093078","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2020,4,27]]}}}