Journal article
Generation of autologous tumor-specific T cells for adoptive transfer based on vaccination, in vitro restimulation and CD3/CD28 dynabead-induced T cell expansion
Department of Hematology 54P4, Center for Cancer Immune Therapy, University Hospital Herlev, Herlev, Denmark.1
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® ClinExVivo™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® CD3/CD28 and CD3/CD28/CD137 T-cell expander, for optimized expansion of tumor-specific T cells. A ratio of 1:3 of Dynabeads® 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®-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.
Language: | English |
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Publisher: | Springer-Verlag |
Year: | 2012 |
Pages: | 1221-1231 |
ISSN: | 14320851 and 03407004 |
Types: | Journal article |
DOI: | 10.1007/s00262-011-1199-8 |
ORCIDs: | Svane, Inge Marie |
Adoptive T-cell therapy CD28 Antigens CD3 Complex Cancer Research Cancer Vaccines Cell Culture Techniques Clinical Trials as Topic Dynabeads ClinExVivo CD3/CD28 Flow Cytometry Humans Immunology Immunotherapy, Adoptive Medicine & Public Health Oncology T-Lymphocytes T-cell expansion Tumor-specific T cells