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" |
