Background information

Downstream applications

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• Figure 1

  BALB/c mice were immunized i.p. with hen eggwhite lysozyme (HEL) in incomplete Freund's adjuvant and pertussis toxin. On day 21 after immunization, mouse spleen cells were restimulated in vitro with HEL for 15 hours. The responding cells were stained and enriched according to secretion of IFN-γ using the Mouse IFN-γ Secretion Assay - Cell Enrichment and Detection Kit. In the stimulated sample, 693 IFN-γ- secreting CD4 + T cells were enriched per 10 6 CD4 + T cells using MS Columns and a MiniMACS™ Separator. In the unstimulated control sample, only 67 IFN-γ-secreting CD4 + T cells were enriched per 10 6 CD4 + T cells.

  Stimulated sample

  Before enrichment	After enrichment
  Unstimulated control

  Before enrichment	After enrichment
  * Percentage represents frequency among CD4 + T cells.

  ** Percentage represents frequency among enriched cells.

• Figure 2

  In this example, BALB/c mice were immunized i.p. with KLH (keyhole limpet hemocyanin) in incomplete Freund's adjuvant and pertussis toxin. On day 21 after immunization, mouse spleen cells were restimulated in vitro with KLH for 15 hours. The responding cells were stained for co-expression of cytokines using the Mouse IFN-γ Secretion Assay - Detection Kit (APC) and Mouse IL-2 Secretion Assay - Detection Kit (PE). The plots show co-expression of IFN-γ- and IL-2-secreting cells gated on viable CD4 + T cells on the stimulated sample as well as on the unstimulated control.

  Stimulated sample	Unstimulated control
  * Percentage represents frequency among CD4 + T cells.

• Selected references

  1. Assenmacher et al. (1998) Sequential production of IL-2, IFN-gamma and IL-10 by individual staphylococcal enterotoxin B-activated T helper lymphocytes. Eur. J. Immunol. 28: 1534-1543
  2. Wu, C. Y. et al. (2002) Distinct lineages of T(H)1 cells have differential capacities for memory cell generation in vivo. Nat. Immunol. 3: 852-858
  3. Zhang, Y. et al. (2001) Interferon gamma stabilizes the T helper cell type 1 phenotype. J. Exp. Med. 194: 165-172
  4. Becker, C. et al. (2001) Adoptive tumor therapy with T lymphocytes enriched through an IFN-gamma capture assay. Nat. Med. 7: 1159-1162
  5. Riberdy, J. M. et al. (2001) Cutting edge: culture with high doses of viral peptide induces previously unprimed CD8(+) T cells to produce cytokine. J. Immunol. 167: 2437-2440
  6. Hayakawa et al. (2001) Critical contribution of IFN-gamma and NK cells, but not perforin-mediated cytotoxicity, to anti-metastatic effect of alpha-galactosylceramide. Eur. J. Immunol. 31: 1720-1727
  7. Yang, J. Q. et al. (2003) Immunoregulatory role of CD1d in the hydrocarbon oil-induced model of lupus nephritis. J. Immunol. 171: 2142-2153
  8. Morris, S. C. et al. (2006)
     IL-4 induces

     in vivo

     production of IFN-gamma by NK and NKT cells.
     J. Immunol. 176: 5299-5305
  9. Berenzon, D. et al. (2003)
     Protracted protection to

     Plasmodium berghei

     malaria is linked to functionally and phenotypically heterogeneous liver memory CD8

     ⁺

     T cells.
     J. Immunol. 171: 2024-2034
  10. Mc Allister, F. et al. (2004)
      T cytotoxic-1 CD8

      ⁺

      T cells are effector cells against pneumocystis in mice.
      J. Immunol. 172: 1132-1138