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Ex vivo expansion of polyclonal and antigen-specific cytotoxic T lymphocytes by artificial APCs expressing ligands for the T-cell receptor, CD28 and 4-1BB

Nature Biotechnology volume 20, pages 143–148 (2002)Cite this article

Abstract

The ex vivo priming and expansion of human cytotoxic T lymphocytes (CTLs) has potential for use in immunotherapy applications for cancer and infectious diseases. To overcome the difficulty in obtaining sufficient numbers of CTLs, we have developed artificial antigen-presenting cells (aAPCs) expressing ligands for the T-cell receptor (TCR) and the CD28 and 4-1BB co-stimulatory surface molecules. These aAPCs reproducibly activate and rapidly expand polyclonal or antigen-specific CD8+ T cells. The starting repertoire of CD8+ T cells was preserved during culture. Furthermore, apoptosis of cultured CD8+ T cells was diminished by this approach. This approach may have important therapeutic implications for adoptive immunotherapy.

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Figure 1: Construction of artificial APCs (aAPCs) from the parental K562 cell line.
Figure 2: Long-term growth of primary polyclonal human CD8+ T cells stimulated with aAPCs in the absence of exogenous cytokines.
Figure 3: Propagation of antigen-specific cytotoxic T cells from an HLA A*0201 donor using K32/4-1BBL aAPCs.
Figure 4: Maintenance of the TCR Vβ repertoire in polyclonal CD8+ T cells after expansion with K32/4-1BBL aAPCs.
Figure 5: Expression of genes involved in T-cell growth and survival after stimulation with aAPCs.
Figure 6: Distinct effects on apoptosis in cultures of polyclonal human CD8+ T cells stimulated with various aAPCs.

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Acknowledgements

We thank William DeMuth for flow sorting, Patricia Rivers and Alanna Toll for technical help, and Coral Hass for administrative excellence. MHC tetramers were supplied by the NIAID Tetramer Facility and the NIH AIDS Research and Reference Reagent Program. Peptide synthesis was provided by the Protein Chemistry Laboratory of the Medical School of the University of Pennsylvania supported by core grants of the Diabetes and Cancer Centers (DK-19525 and CA-16520). M.V.M. was supported in part by training grant DK07748 from the NIH; A.K.T. was supported in part by the Dr. Mildred-Scheel-Stiftung Deutsche Krebschilfe Foundation Grant.

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  1. Abramson Family Cancer Research Institute at the University of Pennsylvania Cancer Center, Philadelphia, 19104, PA

    Marcela V. Maus, Anna K. Thomas, David Allman, Katia Schlienger, James L. Riley & Carl H. June

  2. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, 19104, PA

    Debra G.B. Leonard, David Allman, James L. Riley & Carl H. June

  3. Molecular Diagnostic Core, University of Pennsylvania, Philadelphia, 19104, PA

    Debra G.B. Leonard & Kathakali Addya

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Correspondence to Carl H. June.

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Maus, M., Thomas, A., Leonard, D. et al. Ex vivo expansion of polyclonal and antigen-specific cytotoxic T lymphocytes by artificial APCs expressing ligands for the T-cell receptor, CD28 and 4-1BB. Nat Biotechnol 20, 143–148 (2002). https://doi.org/10.1038/nbt0202-143

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