Abstract
Gamma delta (γδ) T cells and cytokine-induced killer (CIK) cells, which are a heterogeneous population of T lymphocytes and natural killer T (NKT) cells, have been separately expanded ex vivo and shown to be capable of targeting and mediating cytotoxicity against various tumor cells in a major histocompatibility complex-unrestricted manner. However, the co-expansion and co-administration of these immune cells have not been explored. In this study we describe an efficient method to expand simultaneously both CIK and Vγ9Vδ2 T cells, termed as CIKZ cells, from human peripheral blood mononuclear cells (PBMCs) using Zometa, interferon-gamma (IFN-γ), interleukin 2 (IL-2), anti-CD3 antibody and engineered K562 feeder cells expressing CD64, CD137L and CD86. A 21-day culture of PBMCs with this method yielded nearly 20,000-fold expansion of CIKZ cells with γδ T cells making up over 20% of the expanded population. The expanded CIKZ cells exhibited antitumor cytotoxicity and could be modified to express anti-CD19 chimeric antigen receptor (CAR), anti-CEA CAR, and anti-HER2 CAR to enhance their specificity and cytotoxicity against CD19-, CEA-, or HER2-positive tumor cells. The tumor inhibitory activity of anti-CD19 CAR-modified CIKZ cells was further demonstrated in vivo in a Raji tumor mouse model. The findings herein substantiate the feasibility of co-expanding CIK and γδ cells for adoptive cellular immunotherapy applications such as CAR T-cell therapy against cancer.
MeSH terms
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Animals
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Antigens, CD19 / genetics
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Antigens, CD19 / immunology
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B7-2 Antigen / genetics
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B7-2 Antigen / immunology
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Burkitt Lymphoma / immunology
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Burkitt Lymphoma / mortality
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Burkitt Lymphoma / pathology
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Burkitt Lymphoma / therapy*
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Carcinoembryonic Antigen / genetics
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Carcinoembryonic Antigen / immunology
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Cell Proliferation
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Cytokine-Induced Killer Cells / cytology
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Cytokine-Induced Killer Cells / immunology*
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Cytokine-Induced Killer Cells / transplantation
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Cytotoxicity, Immunologic
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Feeder Cells / cytology
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Feeder Cells / immunology
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Gene Expression
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Humans
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Immunotherapy, Adoptive / methods*
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K562 Cells
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Mice
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Primary Cell Culture
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Receptor, ErbB-2 / genetics
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Receptor, ErbB-2 / immunology
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Receptors, Antigen, T-Cell, gamma-delta / genetics
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Receptors, Antigen, T-Cell, gamma-delta / immunology*
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Receptors, IgG / genetics
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Receptors, IgG / immunology
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology*
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Survival Analysis
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T-Lymphocytes / cytology
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T-Lymphocytes / immunology*
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T-Lymphocytes / transplantation
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Tumor Necrosis Factor Receptor Superfamily, Member 9 / genetics
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Tumor Necrosis Factor Receptor Superfamily, Member 9 / immunology
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Xenograft Model Antitumor Assays
Substances
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Antigens, CD19
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B7-2 Antigen
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CD19 molecule, human
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Carcinoembryonic Antigen
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Receptors, Antigen, T-Cell, gamma-delta
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Receptors, IgG
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Recombinant Fusion Proteins
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Tumor Necrosis Factor Receptor Superfamily, Member 9
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ERBB2 protein, human
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Receptor, ErbB-2
Grants and funding
Funding that supported this study was provided to SW from the National Medical Research Council - NMRC/CIRG/1367/2013, NMRC/CIRG/1406/2014 (
http://www.nmrc.gov.sg/index.html), Institute of Bioengineering and Nanotechnology (Biomedical Research Council, Agency for Science, Technology and Research, Singapore;
http://www.ibn.a-star.edu.sg/), and Tessa Therapeutics, Pte Ltd. Singapore (
http://www.tessatherapeutics.com/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.