Co-Expansion of Cytokine-Induced Killer Cells and Vγ9Vδ2 T Cells for CAR T-Cell Therapy

PLoS One. 2016 Sep 6;11(9):e0161820. doi: 10.1371/journal.pone.0161820. eCollection 2016.

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

  • Animals
  • Antigens, CD19 / genetics
  • Antigens, CD19 / immunology
  • B7-2 Antigen / genetics
  • B7-2 Antigen / immunology
  • Burkitt Lymphoma / immunology
  • Burkitt Lymphoma / mortality
  • Burkitt Lymphoma / pathology
  • Burkitt Lymphoma / therapy*
  • Carcinoembryonic Antigen / genetics
  • Carcinoembryonic Antigen / immunology
  • Cell Proliferation
  • Cytokine-Induced Killer Cells / cytology
  • Cytokine-Induced Killer Cells / immunology*
  • Cytokine-Induced Killer Cells / transplantation
  • Cytotoxicity, Immunologic
  • Feeder Cells / cytology
  • Feeder Cells / immunology
  • Gene Expression
  • Humans
  • Immunotherapy, Adoptive / methods*
  • K562 Cells
  • Mice
  • Primary Cell Culture
  • Receptor, ErbB-2 / genetics
  • Receptor, ErbB-2 / immunology
  • Receptors, Antigen, T-Cell, gamma-delta / genetics
  • Receptors, Antigen, T-Cell, gamma-delta / immunology*
  • Receptors, IgG / genetics
  • Receptors, IgG / immunology
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology*
  • Survival Analysis
  • T-Lymphocytes / cytology
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / transplantation
  • Tumor Necrosis Factor Receptor Superfamily, Member 9 / genetics
  • Tumor Necrosis Factor Receptor Superfamily, Member 9 / immunology
  • Xenograft Model Antitumor Assays

Substances

  • Antigens, CD19
  • B7-2 Antigen
  • CD19 molecule, human
  • Carcinoembryonic Antigen
  • Receptors, Antigen, T-Cell, gamma-delta
  • Receptors, IgG
  • Recombinant Fusion Proteins
  • Tumor Necrosis Factor Receptor Superfamily, Member 9
  • ERBB2 protein, human
  • Receptor, ErbB-2

Grant support

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.