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Manufacture of gene modified human T cells with a memory stem/ central memory phenotype

Authors

  • R. Gomez-Eerland
  • B. Nuijen
  • B. Heemskerk
  • N. van Rooij
  • J.H. van den Berg
  • J.H. Beijnen
  • W. Uckert
  • P. Kvistborg
  • T. Schumacher
  • J.B.A.G. Haanen
  • A. Jorritsma

Journal

  • Human Gene Therapy Methods

Citation

  • Hum Gene Ther Methods 25 (5): 277-287

Abstract

  • Advances in genetic engineering have made it possible to generate human T cell products that carry desired functionalities, such as the ability to recognize cancer cells. The currently used strategies for the generation of gene-modified T cell products lead to highly differentiated cells within the infusion product, and on the basis of data obtained in preclinical models, this is likely to impact the efficacy of these products. We set out to develop a Good Manufacturing Practice (GMP) protocol that yields TCR gene modified T cells with more favourably properties for clinical application. Here, we show the robust clinical scale production of human peripheral blood T cells with an early memory phenotype that express a MART-1-specific T cell receptor. By combining selection and stimulation using anti-CD3/CD28 beads for retroviral transduction, followed by expansion in the presence of IL-7 and IL-15, production of a well-defined clinical scale TCR gene modified T cell product could be achieved. A major fraction of the T cells generated in this fashion were shown to co-express CD62L and CD45RA, and express CD27 and CD28 indicating a central memory or memory stem-like phenotype. Furthermore, these cells produced IFN{gamma}, TNF{alpha} and IL-2, and displayed cytolytic activity against target cells expressing the relevant antigen. The T cell products manufactured by this robust and validated GMP production process are now undergoing testing in a phase I/IIa clinical trial in HLA-A*02:01 - MART-1 positive advanced stage melanoma patients. To our knowledge, this is the first clinical trial protocol in which the combination of IL-7 and IL-15 has been applied for the generation of gene-modified T cell products.


DOI

doi:10.1089/hgtb.2014.004