title: Large-scale expansion of γδ T cells and peptide-specific cytotoxic T cells using zoledronate for adoptive immunotherapy.
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.
authors.affiliation: Division of Cancer Immunotherapy, Exploratory Oncology Research and Clinical Trial Center, National Cancer Center, Kashiwa 277-8577, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Yoshikawa
authors.affiliation: Medinet Medical Institute, Setagaya-ku, Tokyo 158-0096, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Takahara
authors.affiliation: Medinet Medical Institute, Setagaya-ku, Tokyo 158-0096, Japan.
authors.forename: Mai
authors.initials: M
authors.lastname: Tomiyama
authors.affiliation: Medinet Medical Institute, Setagaya-ku, Tokyo 158-0096, Japan.
authors.forename: Mie
authors.initials: M
authors.lastname: Nieda
authors.affiliation: Medinet Medical Institute, Setagaya-ku, Tokyo 158-0096, Japan.
authors.forename: Ryuji
authors.initials: R
authors.lastname: Maekawa
authors.affiliation: Division of Cancer Immunotherapy, Exploratory Oncology Research and Clinical Trial Center, National Cancer Center, Kashiwa 277-8577, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Nakatsura
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000276:immunology; D003587:Cytomegalovirus / Q000276:immunology; D004164:Diphosphonates / Q000008:administration & dosage; D053673:Glypicans / Q000276:immunology*; D006801:Humans; D007093:Imidazoles / Q000008:administration & dosage; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D051379:Mice; D009363:Neoplasm Proteins / Q000276:immunology; D009369:Neoplasms / Q000276:immunology* / Q000473:pathology / Q000628:therapy; D010455:Peptides / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D000077211:Zoledronic Acid
chemical_list: D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; D004164:Diphosphonates; D053673:Glypicans; D007093:Imidazoles; D009363:Neoplasm Proteins; D010455:Peptides; C476587:Taa1 protein, mouse; D000077211:Zoledronic Acid