Generation of autologous tumor-specific T cells for adoptive transfer based on vaccination, in vitro restimulation and CD3/CD28 dynabead-induced T cell expansion

Marie Klinge Brimnes, Anne Ortved Gang, Marco Donia, Per Thor Straten, Inge Marie Svane, Sine Reker Hadrup

Research output: Contribution to journalJournal articleResearchpeer-review


Adoptive cell transfer (ACT) of in vitro expanded autologous tumor-infiltrating lymphocytes (TIL) has been shown to exert therapeutic efficacy in melanoma patients. We aimed to develop an ACT protocol based on tumor-specific T cells isolated from peripheral blood and in vitro expanded by Dynabeads(A (R)) ClinExVivo (TM) CD3/CD28. We show here that the addition of an in vitro restimulation step with relevant peptides prior to bead expansion dramatically increased the proportion of tumor-specific T cells in PBMC-cultures. Importantly, peptide-pulsed dendritic cells (DCs) as well as allogeneic tumor lysate-pulsed DCs from the DC vaccine preparation could be used with comparable efficiency to peptides for in vitro restimulation, to increase the tumor-specific T-cell response. Furthermore, we tested the use of different ratios and different types of Dynabeads(A (R)) CD3/CD28 and CD3/CD28/CD137 T-cell expander, for optimized expansion of tumor-specific T cells. A ratio of 1:3 of Dynabeads(A (R)) CD3/CD28 T-cell expander to T cells resulted in the maximum number of tumor-specific T cells. The addition of CD137 did not improve functionality or fold expansion. Both T-cell expansion systems could generate tumor-specific T cells that were both cytotoxic and effective cytokine producers upon antigen recognition. Dynabeads(A (R))-expanded T-cell cultures shows phenotypical characteristics of memory T cells with potential to migrate and expand in vivo. In addition, they possess longer telomeres compared to TIL cultures. Taken together, we demonstrate that in vitro restimulation of tumor-specific T cells prior to bead expansion is necessary to achieve high numbers of tumor-specific T cells. This is effective and easily applicable in combination with DC vaccination, by use of vaccine-generated DCs, either pulsed with peptide or tumor-lysate.
Original languageEnglish
JournalCancer Immunology, Immunotherapy
Issue number8
Pages (from-to)1221-1231
Number of pages11
Publication statusPublished - 2012
Externally publishedYes


  • Cancer Research
  • Oncology
  • Immunology
  • Immunology and Allergy
  • Adoptive T-cell therapy
  • Dynabeads ClinExVivo CD3/CD28
  • T-cell expansion
  • Tumor-speciWc T cells
  • CD137 antigen
  • CD28 antigen
  • CD3 antigen
  • dendritic cell vaccine
  • adoptive transfer
  • antigen recognition
  • article
  • cell expansion
  • cell isolation
  • cell specificity
  • cell transfer
  • cellular immunity
  • clinical protocol
  • controlled study
  • cytokine production
  • dendritic cell
  • human
  • human cell
  • immunostimulation
  • in vitro study
  • in vivo study
  • lymphocyte count
  • lymphocyte migration
  • memory T lymphocyte
  • peripheral blood mononuclear cell
  • priority journal
  • T lymphocyte activation
  • telomere
  • vaccination
  • Antigens, CD28
  • Antigens, CD3
  • Cancer Vaccines
  • Cell Culture Techniques
  • Clinical Trials as Topic
  • Flow Cytometry
  • Humans
  • Immunotherapy, Adoptive
  • T-Lymphocytes
  • Tumor-specific T cells
  • Primates Mammalia Vertebrata Chordata Animalia (Animals, Chordates, Humans, Mammals, Primates, Vertebrates) - Hominidae [86215] human common
  • CD137
  • CD28
  • CD3
  • cytokine
  • peptide
  • 02506, Cytology - Animal
  • 02508, Cytology - Human
  • 10006, Clinical biochemistry - General methods and applications
  • 10064, Biochemistry studies - Proteins, peptides and amino acids
  • 12512, Pathology - Therapy
  • 15002, Blood - Blood and lymph studies
  • 15004, Blood - Blood cell studies
  • 17002, Endocrine - General
  • 18506, Integumentary system - Pathology
  • 24003, Neoplasms - Immunology
  • 24004, Neoplasms - Pathology, clinical aspects and systemic effects
  • 24008, Neoplasms - Therapeutic agents and therapy
  • 34502, Immunology - General and methods
  • 34508, Immunology - Immunopathology, tissue immunology
  • Clinical Chemistry
  • Clinical Immunology
  • Dermatology
  • Methods and Techniques
  • melanoma Melanoma (MeSH) neoplastic disease, integumentary system disease therapy, prevention and control
  • allogeneic tumor lysate
  • Allied Medical Sciences
  • Human Medicine, Medical Sciences
  • dendritic cell immune system
  • peripheral blood mononuclear cell PBMC immune system, blood and lymphatics
  • T cell immune system, blood and lymphatics
  • adoptive cell transfer method ACT method therapeutic and prophylactic techniques, clinical techniques
  • Dynabeads ClinExVivo medical equipment
  • vaccine therapy therapeutic and prophylactic techniques, clinical techniques


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