Activating and propagating polyclonal gamma delta T cells with broad specificity for malignancies

Clin Cancer Res. 2014 Nov 15;20(22):5708-19. doi: 10.1158/1078-0432.CCR-13-3451. Epub 2014 May 15.

Abstract

Purpose: To activate and propagate populations of γδ T cells expressing polyclonal repertoire of γ and δ T-cell receptor (TCR) chains for adoptive immunotherapy of cancer, which has yet to be achieved.

Experimental design: Clinical-grade artificial antigen-presenting cells (aAPC) derived from K562 tumor cells were used as irradiated feeders to activate and expand human γδ T cells to clinical scale. These cells were tested for proliferation, TCR expression, memory phenotype, cytokine secretion, and tumor killing.

Results: γδ T-cell proliferation was dependent upon CD137L expression on aAPC and addition of exogenous IL2 and IL21. Propagated γδ T cells were polyclonal as they expressed TRDV1, TRDV2-2, TRDV3, TRDV5, TRDV7, and TRDV8 with TRGV2, TRGV3F, TRGV7, TRGV8, TRGV9*A1, TRGV10*A1, and TRGV11 TCR chains. IFNγ production by Vδ1, Vδ2, and Vδ1(neg)Vδ2(neg) subsets was inhibited by pan-TCRγδ antibody when added to cocultures of polyclonal γδ T cells and tumor cell lines. Polyclonal γδ T cells killed acute and chronic leukemia, colon, pancreatic, and ovarian cancer cell lines, but not healthy autologous or allogeneic normal B cells. Blocking antibodies demonstrated that polyclonal γδ T cells mediated tumor cell lysis through combination of DNAM1, NKG2D, and TCRγδ. The adoptive transfer of activated and propagated γδ T cells expressing polyclonal versus defined Vδ TCR chains imparted a hierarchy (polyclonal>Vδ1>Vδ1(neg)Vδ2(neg)>Vδ2) of survival of mice with ovarian cancer xenografts.

Conclusions: Polyclonal γδ T cells can be activated and propagated with clinical-grade aAPCs and demonstrate broad antitumor activities, which will facilitate the implementation of γδ T-cell cancer immunotherapies in humans.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adoptive Transfer
  • Animals
  • Antigen-Presenting Cells / immunology
  • Antigen-Presenting Cells / metabolism
  • Antigens, Differentiation, T-Lymphocyte / metabolism
  • Cell Culture Techniques
  • Cell Line, Tumor
  • Cytokines / biosynthesis
  • Cytokines / pharmacology
  • Disease Models, Animal
  • Gene Expression
  • Humans
  • Interferon-gamma / biosynthesis
  • Lymphocyte Activation / immunology*
  • Mice
  • Mice, Transgenic
  • NK Cell Lectin-Like Receptor Subfamily K / metabolism
  • Neoplasms / immunology*
  • Neoplasms / metabolism*
  • Neoplasms / mortality
  • Neoplasms / therapy
  • RNA, Messenger
  • Receptors, Antigen, T-Cell, gamma-delta / genetics
  • Receptors, Antigen, T-Cell, gamma-delta / metabolism*
  • T-Cell Antigen Receptor Specificity / immunology*
  • T-Lymphocyte Subsets / drug effects
  • T-Lymphocyte Subsets / immunology*
  • T-Lymphocyte Subsets / metabolism*
  • Xenograft Model Antitumor Assays

Substances

  • Antigens, Differentiation, T-Lymphocyte
  • CD226 antigen
  • Cytokines
  • KLRK1 protein, human
  • NK Cell Lectin-Like Receptor Subfamily K
  • RNA, Messenger
  • Receptors, Antigen, T-Cell, gamma-delta
  • Interferon-gamma