Optimization and validation of a robust human T-cell culture method for monitoring phenotypic and polyfunctional antigen-specific CD4 and CD8 T-cell responses

Cytotherapy. 2009;11(7):912-22. doi: 10.3109/14653240903136987.


Background aims: Monitoring cellular immune responses is one prerequisite for the rational development of cancer vaccines.

Methods: We describe an extensive effort to optimize and validate quantitatively an in vitro T-cell culture method by determining the phenotype and function of both CD4(+) and CD8(+) T cells, including measurement of the phenotype markers CCR7, CD45RA, CD28 and CD27 and the functional markers interferon (IFN)-gamma, interleukin (IL)-2, macrophage inflammatory protein (MIP)-1beta, tumor necrosis factor (TNF)-alpha and CD107a.

Results: Autologous peripheral blood mononuclear cells (PBMC) were potent stimulators that expanded antigen (Ag)-specific CD8(+) T cells during short-term culture with the addition of IL-2 and IL-15 cytokines. Polyfunctional Ag-specific CD4(+) and CD8(+) T cells were detectable using this method.

Conclusions: Our culture system represents a robust human T-cell culture protocol that permits phenotypic, quantitative and qualitative evaluation of vaccine-induced CD4(+) and CD8(+) T-cell responses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens / immunology
  • Antigens, CD / metabolism
  • Antigens, Differentiation / metabolism
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / metabolism*
  • CD4-Positive T-Lymphocytes / pathology
  • CD8-Positive T-Lymphocytes / immunology
  • CD8-Positive T-Lymphocytes / metabolism*
  • CD8-Positive T-Lymphocytes / pathology
  • Cancer Vaccines
  • Cell Culture Techniques*
  • Cells, Cultured
  • Cytokines / metabolism
  • Feasibility Studies
  • Humans
  • Neoplasms / immunology*
  • Neoplasms / therapy


  • Antigens
  • Antigens, CD
  • Antigens, Differentiation
  • Cancer Vaccines
  • Cytokines