Background information

Downstream applications

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• Figure 1

  PBMCs from a CMV + donor were stimulated with PepTivator™ CMV pp65 or left untreated. Cells were stained and isolated according to secretion of IFN-γ using the IFN-γ Secretion Assay - Cell Enrichment and Detection Kit. The cells were counterstained with CD8-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and PI fluorescence.

  Stimulated sample

  Before enrichment	After enrichment
  Unstimulated control

  Before enrichment	After enrichment

• Selected references

  1. Brosterhus, H. et al. (1999) Enrichment and detection of live antigen-specific CD4(+) and CD8(+) T cells based on cytokine secretion. Eur. J. Immunol. 29: 4053-4059
  2. Bissinger, A. L. et al. (2002) Isolation and expansion of human cytomegalovirus- specific cytotoxic T lymphocytes using interferon-gamma secretion assay. Exp. Hematol. 30: 1178-1184
  3. Bitmansour, A. D. et al. (2002)
     Direct

     ex vivo

     analysis of human CD4(+) memory T cell activation requirements at the single clonotype level.
     J. Immunol. 169: 1207-1218
  4. Bickham, K. et al. (2001)
     EBNA1-specific CD4

     ⁺

     T cells in healthy carriers of Epstein-Barr virus are primarily Tʜ1 in function.
     J. Clin. Invest. 107: 121-130
  5. Cohen, G. B. et al. (2002) Clonotype tracking of TCR repertoires during chronic virus infections. Virology 304: 474-484
  6. Koehne, G. et al. (2002) Quantitation, selection, and functional characterization of Epstein-Barr virus-specific and alloreactive T cells detected by intracellular interferon-gamma production and growth of cytotoxic precursors. Blood 99: 1730-1740
  7. Altfeld, M. et al. (2001) Vpr is preferentially targeted by CTL during HIV-1 infection. J. Immunol. 167: 2743-2752
  8. Douek, D. C. et al. (2002)
     HIV preferentially infects HIV-specific CD4

     ⁺

     T cells.
     Nature 417: 95-98
  9. Lichterfeld, M. et al. (2004)
     HIV-1-specific cytotoxicity is preferentially mediated by a subset of CD8

     ⁺

     T cells producing both interferon-gamma and tumor necrosis factor-alpha.
     Blood 104: 487-494
  10. Lee, S. K. et al. (2002) The functional CD8 T cell response to HIV becomes type-specific in progressive disease. J. Clin. Invest. 110: 1339-1347
  11. Desombere, I. et al. (2003) The interferon gamma secretion assay: a reliable tool to study interferon gamma production at the single cell level. J. Immunol. Methods 286: 167-185
  12. Meidenbauer, N. et al. (2003) Survival and tumor localization of adoptively transferred Melan-A-specific T cells in melanoma patients. J. Immunol. 170: 2161-2169
  13. Ayyoub, M. et al. (2004)
      An immunodominant SSX-2-derived epitope recognized by CD4

      ⁺

      T cells in association with HLA-DR.
      J. Clin. Invest. 113: 1225-1233
  14. Pittet, M. J. et al. (2001)
      Ex vivo

      IFN-gamma secretion by circulating CD8 T lymphocytes: implications of a novel approach for T cell monitoring in infectious and malignant diseases.
      J. Immunol. 166: 7634-7640
  15. Märten, A. et al. (2002) Generation of activated and antigen-specific T cells with cytotoxic activity after co-culture with dendritic cells. Cancer Immunol. Immunother. 51: 25-32
  16. Meyer zu Büschenfelde, C. M. et al. (2001) The generation of both T killer and Tʜ cell clones specific for the tumor-associated antigen HER2 using retrovirally transduced dendritic cells. J. Immunol. 167: 1712-1719
  17. Becker, C. et al. (2001) Adoptive tumor therapy with T lymphocytes enriched through an IFN-gamma capture assay. Nat. Med. 7: 1159-1162
  18. Novak, E. J. et al. (2001)
      Tetramer-guided epitope mapping: rapid identification and characterization of immunodominant CD4

      ⁺

      T cell epitopes from complex antigens.
      J. Immunol. 166: 6665-6670
  19. Deniz, G. et al. (2002) Human NK1 and NK2 subsets determined by purification of IFN-gamma-secreting and IFN-gamma-nonsecreting NK cells. Eur. J. Immunol. 32: 879-884
  20. Akdis, M. et al. (2004) Immune responses in healthy and allergic individuals are characterized by a fine balance between allergen-specific T regulatory 1 and T helper 2 cells. J. Exp. Med. 199: 1567-1575
  21. Meyer-Olson, D. et al. (2003) Analysis of the TCR beta variable gene repertoire in chimpanzees: identification of functional homologs to human pseudogenes. J. Immunol. 170: 4161-4169
  22. Gutzmer, R. et al. (2003) Human dendritic cells express the IL-18R and are chemoattracted to IL-18. J. Immunol. 171: 6363-6371
  23. Feuchtinger T. et al. (2004)
      Isolation and expansion of human adenovirus-specific CD4

      ⁺

      and CD8

      ⁺

      T cells according to IFN-gamma secretion for adjuvant immunotherapy.
      Exp. Hematol. 32: 282-289

• Customer reports and application notes

  Effector memory T helper cells secrete IFN-γ upon stimulation with cytokines: a role in chronic inflammation. Sattler et al. (2011) MACS&more 13(1).

• Special protocols

  Detection and enrichment of cytokine-secreting cells from whole blood
  Detection and enrichment of cytokine-secreting cells
  Expansion of antigen-specific cytokine-secreting T cells