Mouse IL-10 Secretion Assay – Cell Enrichment and Detection Kit ( #130-090-490 )

Mouse IL-10 Secretion Assay – Cell Enrichment and Detection Kit

Description

The Mouse IL-10 Secretion Assay - Cell Enrichment and Detection Kit was developed for the sensitive detection and the enrichment of murine IL-10-secreting cells.

Details

Background information
Interleukin-10 (IL-10) is predominantly secreted by CD4⁺ T cells and antigen-presenting cells, such as monocytes/macrophages and dendritic cells. IL-10 has important suppressive functions on immune responses and is involved in the maintenance of tolerance. IL-10 blocks activation of cytokine synthesis by T_H1 cells, activated monocytes, and NK cells. It can stimulate the immunoglobulin production by B cells.

Downstream applications
The Mouse IL-10 Secretion Assay was used for the analysis of the sequential production of cytokines by T helper lymphocytes.¹The assay was also used for analysis of IL-10 expression by T_H2 cells.²It has also been used to analyze secretion of IL-10 by dendritic cells from mouse spleen³ and by T cells from bronchioalveolar lavages⁴.

Columns

MS, LS, or autoMACS Columns.

Figure 1

BALB/c mice were immunized and the spleen cells restimulated with HEL as described for the Mouse IFN-γ Secretion Assay. The responding cells were stained and enriched according to secretion of IL-10 using the Mouse IL-10 Secretion Assay - Cell Enrichment and Detection Kit.
In the stimulated sample 144 IL-10-secreting CD4⁺ T cells were enriched per 10⁶ CD4⁺ T cells using MS Columns and a MiniMACS™ Separator. In the unstimulated control sample, only seven IL-10-secreting CD4⁺ T cells were detected per 10⁶ CD4⁺ T cells after enrichment.

Stimulated sample

A: Before enrichment

B: After enrichment

Unstimulated control

C: Before enrichment

D: After enrichment

* Percentage represents frequency among CD4⁺ T cells.
** Percentage represents frequency among enriched cells.

Figure 2

BALB/c mice were immunized i.p. with keyhole limpet hemocyanin (KLH) in incomplete Freund's adjuvant and pertussis toxin. On day 14 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 IL-10 Secretion Assay - Detection Kit (APC) and Mouse IL-2 Secretion Assay - Detection Kit (PE). The plots show IL-10- and IL-2-secreting cells gated on viable CD4⁺ T cells for the stimulated sample as well as for the unstimulated control.

A: Stimulated sample

B: Unstimulated control

* Percentage represents frequency among CD4⁺ T cells.