IL-4 Secretion Assay – Cell Enrichment and Detection Kit (PE), human

Background information

Downstream applications

Columns

• Figure 1

  PBMCs from a CMV + donor were stimulated for 16 hours with CMV lysate. The responding cells were stained and isolated according to secretion of IL-4 using the IL-4 Secretion Assay - Cell Enrichment and Detection Kit. In the stimulated sample, 24 IL-4 secreting CD4 + T cells were enriched per 10 6 CD4 + T cells using MS Columns and a MiniMACS™ Separator. In the unstimulated control sample, two IL-4 secreting CD4 + T cells were detected per 10 6 CD4 + T cells after enrichment. There seemed to be no difference between stimulated and unstimulated sample before enrichment.

  Stimulated sample
  Before enrichment	After enrichment
  Unstimulated control
  Before enrichment	After enrichment
  *Please refer to the data sheet for detailed gating strategy and analysis.

  ** Percentage represents frequency among enriched cells.

• 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. Akdis et al. (2000) MACS&more 4: 9-10
  3. Oelke, M. et al. (2000)
     Functional characterization of CD8

     ⁺

     antigen-specific cytotoxic T lymphocytes after enrichment based on cytokine secretion: comparison with the MHC-tetramer technology.
     Scand. J. Immunol. 52: 544-549
  4. 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
  5. 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
  6. Araki, M. et al. (2002)
     Tʜ2 bias of CD4

     ⁺

     NKT cells derived from multiple sclerosis in remission.
     Int. Immunol. 15: 279-288
  7. Veldman, C. et al. (2003) Dichotomy of autoreactive Th1 and Th2 cell responses to desmoglein 3 in patients with pemphigus vulgaris (PV) and healthy carriers of PV-associated HLA class II alleles. J. Immunol. 170: 635-642
  8. Schwaiger, S. et al. (2003)
     IL-4-producing CD8

     ⁺

     T cells with a CD62L

     ^++(bright)

     phenotype accumulate in a subgroup of older adults and are associated with the maintenance of intact humoral immunity in old age.
     J. Immunol. 170: 613-619

• 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