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


Authors: Lin, Y.; Gallardo, H. F.; Ku, G. Y.; Li, H.; Manukian, G.; Rasalan, T. S.; Xu, Y.; Terzulli, S. L.; Old, L. J.; Allison, J. P.; Houghton, A. N.; Wolchok, J. D.; Yuan, J.
Article Title: Optimization and validation of a robust human T-cell culture method for monitoring phenotypic and polyfunctional antigen-specific CD4 and CD8 T-cell responses
Abstract: 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<sup>+</sup> and CD8<sup>+</sup> T cells, including measurement of the phenotype markers CCR7, CD45RA, CD28 and CD27 and the functional markers interferon (IFN)-γ, interleukin (IL)-2, macrophage inflammatory protein (MIP)-1β, tumor necrosis factor (TNF)-α and CD107a. Results Autologous peripheral blood mononuclear cells (PBMC) were potent stimulators that expanded antigen (Ag)-specific CD8<sup>+</sup> T cells during short-term culture with the addition of IL-2 and IL-15 cytokines. Polyfunctional Ag-specific CD4<sup>+</sup> and CD8<sup>+</sup> 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 <sup>+</sup> and CD8<sup>+</sup> T-cell responses. © 2009 ISCT.
Keywords: controlled study; human cell; neoplasms; cd8+ t lymphocyte; cd8-positive t-lymphocytes; cells, cultured; interleukin 2; macrophage inflammatory protein 1beta; in vitro study; validation study; cytokines; cellular immunity; immune response; tumor necrosis factor alpha; human cell culture; antigens; gamma interferon; cancer vaccines; antigen specificity; feasibility studies; quantitative analysis; ny eso 1 antigen; cd4+ t lymphocyte; cd4-positive t-lymphocytes; cell culture techniques; cytokine production; immunophenotyping; melan a; immunological monitoring; antigens, cd; culture technique; chemokine receptor ccr7; phenotypic variation; cell expansion; recombinant interleukin 7; cd28 antigen; peripheral blood mononuclear cell; lymphocyte function; antigen-specifi c t cell; cd4 t cell; cd8 t cell; immune monitoring,phenotype; polyfunctional analysis; cd27 antigen; cd45ra antigen; lysosome associated membrane protein 1; recombinant interleukin 15; recombinant interleukin 2; tetramer; cell culture monitoring; antigens, differentiation
Journal Title: Cytotherapy
Volume: 11
Issue: 7
ISSN: 1465-3249
Publisher: Elsevier Science Ltd.  
Date Published: 2009-01-01
Start Page: 912
End Page: 922
Language: English
DOI: 10.3109/14653240903136987
PUBMED: 19903103
PROVIDER: scopus
PMCID: PMC2932850
DOI/URL:
Notes: --- - "Cited By (since 1996): 1" - "Export Date: 30 November 2010" - "CODEN: CYTRF" - "Source: Scopus"
Altmetric Score
MSK Authors
  1. Jedd D Wolchok
    669 Wolchok
  2. Hao Li
    7 Li
  3. Yun Lin
    3 Lin
  4. Geoffrey Yuyat Ku
    92 Ku
  5. James P Allison
    129 Allison
  6. Jianda Yuan
    100 Yuan
  7. Alan N Houghton
    273 Houghton
  8. Lloyd J Old
    386 Old
  9. Teresa Rasalan
    25 Rasalan
  10. Yinyan Xu
    11 Xu