Generation of Functional, Antigen-Specific CD8+ Human T Cells from Cord Blood Stem Cells Using Exogenous Notch and Tetramer-TCR Signaling

Authors

  • Irina Fernandez,

    1. Department of Biomedical Engineering, The University of Texas at Austin, Austin, Texas, USA
    2. Dell Pediatric Research Institute and, The University of Texas at Austin, Austin, Texas, USA
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  • Tracy P. Ooi,

    1. Department of Biomedical Engineering, The University of Texas at Austin, Austin, Texas, USA
    2. Section of Molecular Cell and Developmental Biology, The Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, Texas, USA
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  • Krishnendu Roy

    Corresponding author
    1. Department of Biomedical Engineering, The University of Texas at Austin, Austin, Texas, USA
    2. Section of Molecular Cell and Developmental Biology, The Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, Texas, USA
    • Correspondence: Krishnendu Roy, Ph.D., Wallace H. Coulter, Department of Biomedical Engineering at Georgia Tech and Emory University and The Parker H. Petit Institute for Bioengineering & Bioscience, IBB 2314, Georgia Institute of Technology, Atlanta, Georgia, USA. Telephone: +1–404-385–6166; e-mail: krish.roy@gatech.edu

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Abstract

In vitro differentiation of mouse and human stem cells into early T cells has been successfully demonstrated using artificial Notch signaling systems. However, generation of mature, antigen-specific, functional T cells, directly from human stem cells has remained elusive, except when using stromal coculture of stem cells retrovirally transfected with antigen-specific T cell receptors (TCRs). Here we show that human umbilical cord blood (UCB)-derived CD34+CD38−/low hematopoietic stem cells can be successfully differentiated into functional, antigen-specific cytotoxic CD8+ T cells without direct stromal coculture or retroviral TCR transfection. Surface-immobilized Notch ligands (DLL1) and stromal cell conditioned medium successfully induced the development of CD1a+CD7+ and CD4+CD8+ early T cells. These cells, upon continued culture with cytomegalovirus (CMV) or influenza-A virus M1 (GIL) epitope-loaded human leukocyte antigen (HLA)-A*0201 tetramers, resulted in the generation of a polyclonal population of CMV-specific or GIL-specific CD8+ T cells, respectively. Upon further activation with antigen-loaded target cells, these antigen-specific, stem cell-derived T cells exhibited cytolytic functionality, specifically CD107a surface mobilization, interferon gamma (IFNg) production, and Granzyme B secretion. Such scalable, in vitro generation of functional, antigen-specific T cells from human stem cells could eventually provide a readily available cell source for adoptive transfer immunotherapies and also allow better understanding of human T cell development. Stem Cells 2014;32:93–104

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