The IFN-γ Secretion Assays - Cell Enrichment and Detection Kit was developed for the sensitive detection as well as the enrichment of murine IFN-γ-secreting cells.

Data and images for Mouse IFN-γ Secretion Assay – Cell Enrichment and Detection Kit (PE)

Figures

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
View details

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.
View details

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.
Unstimulated control
Before enrichment
After enrichment
View details

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.
View details

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.

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
View details

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.
View details

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.

Specifications for Mouse IFN-γ Secretion Assay – Cell Enrichment and Detection Kit (PE)

Overview

The IFN-γ Secretion Assays - Cell Enrichment and Detection Kit was developed for the sensitive detection as well as the enrichment of murine IFN-γ-secreting cells.

Detailed product information

Background information

Interferon-gamma (IFN-γ) is predominantly secreted by activated CD8
+
and CD4
+
memory and effector T cells as well as by NK cells. It is mainly involved in the regulation of inflammatory immune responses. These T1 types of immune mechanisms are effective against intracellular pathogens and tumors and can be involved in immunological disorders, such as autoimmune reactions.

Downstream applications

The Mouse IFN-γ Secretion Assay was used to analyze the sequential production of cytokines by individual staphylococcal enterotoxin B-activated T helper lymphocytes
1
and for studies in T
H
1 cell differentiation
2,3
.
The Mouse IFN-γ Secretion Assay was also used for isolation of tumor-reactive T cells from immunized mice for adoptive T cell transfer experiments
4
and for analysis of T cells from mice immunized with viral antigens
5
. Murine NKT cells were analyzed
6,7,8
for IFN-γ secretion and isolated
7
after stimulation with glycolipid antigens. IFN-γ expression by T cells was also analyzed for intrahepatic mononuclear cells
9
and cells from bronchioalveolar lavages
10
. The Mouse IFN-γ Secretion Assay can also be combined with peptide-tetramer staining.
8

Columns

MS, LS, or autoMACS
®
Columns.

References for Mouse IFN-γ Secretion Assay – Cell Enrichment and Detection Kit (PE)

Publications

  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

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