Ex vivo expansion protocol for human tumor specific T cells for adoptive T cell therapy

J Immunol Methods. 2010 Apr 15;355(1-2):52-60. doi: 10.1016/j.jim.2010.02.004. Epub 2010 Feb 19.


Adoptive T cell therapy is a promising treatment strategy for patients with different types of cancer. The methods used for generation of high numbers of tumor specific T cells usually require long-term ex vivo culture, which frequently lead to generation of terminally differentiated effector cells, demonstrating low persistence in vivo. Therefore, optimization of protocols for generation of T cells for adoptive cell therapy is warranted. The aim of this work was to develop a protocol for expansion of antigen-specific T cells using Dynabeads CD3/CD28 to obtain T cells expressing markers important for in vivo persistence and survival. To achieve high numbers of antigen-specific T cells following expansion, we have tested the effect of depleting regulatory T cells using Dynabeads CD25 and including a pre-stimulation step with peptide prior to the non-specific expansion with Dynabeads. Our data demonstrate that virus- and tumor specific T cells can be expanded to high numbers using Dynabeads CD3/CD28 following optimization of the culture conditions. The expansion protocol presented here results in enrichment of antigen-specific CD8(+) T cells with an early/intermediate memory phenotype. This is observed even when the antigen-specific CD8(+) T cells demonstrated a terminal effector phenotype prior to expansion. This protocol thus results in expanded T cells with a phenotypic profile which may increase the chance of retaining long-term persistence following adoptive transfer. Based on these data we have developed a cGMP protocol for expansion of tumor specific T cells for adoptive T cell therapy.

Publication types

  • Clinical Trial

MeSH terms

  • Adoptive Transfer*
  • Antigens, CD / immunology*
  • CD8-Positive T-Lymphocytes / immunology*
  • Cell Culture Techniques / methods
  • Cell Proliferation*
  • Cell Survival
  • Female
  • Humans
  • Immunologic Memory
  • Immunomagnetic Separation*
  • Male
  • Neoplasms / immunology
  • Neoplasms / therapy


  • Antigens, CD