Large-scale expansion of γδ T cells and peptide-specific cytotoxic T cells using zoledronate for adoptive immunotherapy

Int J Oncol. 2014 Nov;45(5):1847-56. doi: 10.3892/ijo.2014.2634. Epub 2014 Sep 3.

Abstract

Specific cellular immunotherapy for cancer requires efficient generation and expansion of cytotoxic T lymphocytes (CTLs) that recognize tumor-associated antigens. However, it is difficult to isolate and expand functionally active T-cells ex vivo. In this study, we investigated the efficacy of a new method to induce expansion of antigen-specific CTLs for adoptive immunotherapy. We used tumor-associated antigen glypican-3 (GPC3)-derived peptide and cytomegalovirus (CMV)-derived peptide as antigens. Treatment of human peripheral blood mononuclear cells (PBMCs) with zoledronate is a method that enables large-scale γδ T-cell expansion. To induce expansion of γδ T cells and antigen-specific CTLs, the PBMCs of healthy volunteers or patients vaccinated with GPC3 peptide were cultured with both peptide and zoledronate for 14 days. The expansion of γδ T cells and peptide-specific CTLs from a few PBMCs using zoledronate yields cell numbers sufficient for adoptive transfer. The rate of increase of GPC3‑specific CTLs was approximately 24- to 170,000-fold. These CD8(+) cells, including CTLs, showed GPC3-specific cytotoxicity against SK-Hep-1/hGPC3 and T2 pulsed with GPC3 peptide, but not against SK-Hep-1/vec and T2 pulsed with human immunodeficiency virus peptide. On the other hand, CD8(-) cells, including γδ T cells, showed cytotoxicity against SK-Hep-1/hGPC3 and SK-Hep-1/vec, but did not show GPC3 specificity. Furthermore, adoptive cell transfer of CD8(+) cells, CD8(-) cells, and total cells after expansion significantly inhibited tumor growth in an NOD/SCID mouse model. This study indicates that simultaneous expansion of γδ T cells and peptide-specific CTLs using zoledronate is useful for adoptive immunotherapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Neoplasm / immunology
  • Cell Adhesion Molecules / immunology
  • Cytomegalovirus / immunology
  • Diphosphonates / administration & dosage
  • Glypicans / immunology*
  • Humans
  • Imidazoles / administration & dosage
  • Immunotherapy, Adoptive / methods*
  • Leukocytes, Mononuclear / immunology
  • Mice
  • Neoplasm Proteins / immunology
  • Neoplasms / immunology*
  • Neoplasms / pathology
  • Neoplasms / therapy
  • Peptides / immunology
  • T-Lymphocytes, Cytotoxic / immunology*
  • Zoledronic Acid

Substances

  • Antigens, Neoplasm
  • Cell Adhesion Molecules
  • Diphosphonates
  • Glypicans
  • Imidazoles
  • Neoplasm Proteins
  • Peptides
  • Taa1 protein, mouse
  • Zoledronic Acid