Co-Expansion of Cytokine-Induced Killer Cells and Vγ9Vδ2 T Cells for CAR T-Cell Therapy.
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.
Department of Biological Sciences, National University of Singapore, Singapore, 117543, Singapore.
Shou-Hui
SH
Du
Department of Biological Sciences, National University of Singapore, Singapore, 117543, Singapore.
Zhendong
Z
Li
Tessa Therapeutics, Pte Ltd., Singapore, 239351, Singapore.
Can
C
Chen
Department of Biological Sciences, National University of Singapore, Singapore, 117543, Singapore.|Institute of Bioengineering and Nanotechnology, Singapore, 138669, Singapore.
Wee-Kiat
WK
Tan
Department of Biological Sciences, National University of Singapore, Singapore, 117543, Singapore.
Zhixia
Z
Chi
Tessa Therapeutics, Pte Ltd., Singapore, 239351, Singapore.
Timothy Weixin
http://orcid.org/0000-0003-4029-3533
TW
Kwang
Department of Gastrointestinal Surgery, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510150, China.
Xue-Hu
XH
Xu
Department of Biological Sciences, National University of Singapore, Singapore, 117543, Singapore.|Institute of Bioengineering and Nanotechnology, Singapore, 138669, Singapore.
Shu
S
Wang
D000818:Animals; D018941:Antigens, CD19 / Q000235:genetics / Q000276:immunology; D051940:B7-2 Antigen / Q000235:genetics / Q000276:immunology; D002051:Burkitt Lymphoma / Q000276:immunology / Q000401:mortality / Q000473:pathology / Q000628:therapy*; D002272:Carcinoembryonic Antigen / Q000235:genetics / Q000276:immunology; D049109:Cell Proliferation; D055612:Cytokine-Induced Killer Cells / Q000166:cytology / Q000276:immunology* / Q000637:transplantation; D003602:Cytotoxicity, Immunologic; D061252:Feeder Cells / Q000166:cytology / Q000276:immunology; D015870:Gene Expression; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D020014:K562 Cells; D051379:Mice; D061251:Primary Cell Culture; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000276:immunology; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics / Q000276:immunology*; D017452:Receptors, IgG / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*; D016019:Survival Analysis; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000637:transplantation; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000235:genetics / Q000276:immunology; D023041:Xenograft Model Antitumor Assays
D018941:Antigens, CD19; D051940:B7-2 Antigen; D002272:Carcinoembryonic Antigen; D018719:Erb-b2 Receptor Tyrosine Kinases; D016692:Receptors, Antigen, T-Cell, gamma-delta; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; C000604811:CD19 molecule, human; C508053:ERBB2 protein, human