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| Mouse IFN-γ Secretion Assay – Cell Enrichment and Detection Kit |
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| Description |
| 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. |
| Details |
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 lymphocytes1 and for studies in TH1 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 experiments4 and for analysis of T cells from mice immunized with viral antigens.5 Murine NKT cells were analyzed6,7,8 for IFN-γ secretion and isolated7 after stimulation with glycolipid antigens. IFN-γ expression by T cells was also analyzed for intrahepatic mononuclear cells9 and cells from bronchioalveolar lavages10. The Mouse IFN-γ Secretion Assay can also be combined with peptide-tetramer staining.8 |
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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 106 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 106 CD4+ T cells. |
| Stimulated sample |
| A: Before enrichment |
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| B: After enrichment |
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| Unstimulated control |
| C: Before enrichment |
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| D: After enrichment |
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* Percentage represents frequency among CD4+ T cells.
** Percentage represents frequency among enriched cells. |
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| 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 coexpression of cytokines using the Mouse IFN-γ Secretion Assay - Detection Kit (APC) and Mouse IL-2 Secretion Assay - Detection Kit (PE). The plots show coexpression 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. |
| A: Stimulated sample |
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| B: Unstimulated control |
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| * Percentage represents frequency among CD4+ T cells. |
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| Details |
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| Products |
| Mouse IFN-γ Secretion Assay – Cell Enrichment and Detection Kit (PE) |
- for 50 tests with 107 total cells
Download datasheet
130-090-517
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| References |
| 1. Assenmacher et al. (1998) Eur. J. Immunol. 28: 1534-1543. |
| 2. Wu et al. (2002) Nat. Immunol. 3: 852-858. |
| 3. Zhang et al. (2001) J. Exp. Med. 194: 165-172. |
| 4. Becker et al. (2001) Nature Med. 7: 1159-1162. |
| 5. Riberdy et al. (2001) J. Immunol. 167: 2437-2440. |
| 6. Hayakawa et al. (2001) Eur. J. Immunol. 31: 1720-1727. |
| 7. Yang et al. (2003) J. Immunol. 171: 2142-2153. |
| 8. Morris et al. (2006) J. Immunol. 176: 5299-5305. |
| 9. Berenzon et al. (2003) J. Immunol. 171: 2024-2034. |
| 10. Mc Allister et al. (2004) J. Immunol. 172: 1132-1138. |
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