Human CD8 T cells generated in vitro from hematopoietic stem cells are functionally mature

BMC Immunol. 2011 Mar 23;12:22. doi: 10.1186/1471-2172-12-22.

Abstract

Background: T cell development occurs within the highly specialized thymus. Cytotoxic CD8 T cells are critical in adaptive immunity by targeting virally infected or tumor cells. In this study, we addressed whether functional CD8 T cells can be generated fully in vitro using human umbilical cord blood (UCB) hematopoietic stem cells (HSCs) in coculture with OP9-DL1 cells.

Results: HSC/OP9-DL1 cocultures supported the differentiation of CD8 T cells, which were TCR/CD3(hi) CD27(hi) CD1a(neg) and thus phenotypically resembled mature functional CD8 single positive thymocytes. These in vitro-generated T cells also appeared to be conventional CD8 cells, as they expressed high levels of Eomes and low levels of Plzf, albeit not identical to ex vivo UCB CD8 T cells. Consistent with the phenotypic and molecular characterization, upon TCR-stimulation, in vitro-generated CD8 T cells proliferated, expressed activation markers (MHC-II, CD25, CD38), secreted IFN-γ and expressed Granzyme B, a cytotoxic T-cell effector molecule.

Conclusion: Taken together, the ability to direct human hematopoietic stem cell or T-progenitor cells towards a mature functional phenotype raises the possibility of establishing cell-based treatments for T-immunodeficiencies by rapidly restoring CD8 effector function, thereby mitigating the risks associated with opportunistic infections.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / biosynthesis
  • CD8-Positive T-Lymphocytes / cytology
  • CD8-Positive T-Lymphocytes / immunology
  • CD8-Positive T-Lymphocytes / metabolism*
  • Cell Line
  • Coculture Techniques
  • Fetal Blood / cytology
  • Gene Expression Regulation, Developmental / immunology
  • Granzymes / genetics
  • Granzymes / metabolism
  • Hematopoietic Stem Cells / cytology
  • Humans
  • Immunophenotyping
  • Interferon-gamma / genetics
  • Interferon-gamma / metabolism
  • Kruppel-Like Transcription Factors / genetics
  • Kruppel-Like Transcription Factors / immunology
  • Kruppel-Like Transcription Factors / metabolism*
  • Lymphocyte Activation*
  • Lymphopoiesis*
  • Promyelocytic Leukemia Zinc Finger Protein
  • Receptors, Antigen, T-Cell / immunology
  • Stromal Cells / cytology
  • T-Box Domain Proteins / genetics
  • T-Box Domain Proteins / immunology
  • T-Box Domain Proteins / metabolism*

Substances

  • Antigens, CD
  • EOMES protein, human
  • Kruppel-Like Transcription Factors
  • Promyelocytic Leukemia Zinc Finger Protein
  • Receptors, Antigen, T-Cell
  • T-Box Domain Proteins
  • ZBTB16 protein, human
  • Interferon-gamma
  • Granzymes