Intracellular Staining by Flow Cytometry
|Detection of IL‑27/IL‑35 EBI3 Subunit in IL-18-treated KG‑1 Human Cell Line by Flow Cytometry. KG‑1 human acute myelogenous leukemia cell line, resting (open histogram) or treated with rhIL-18 (filled histogram) was stained with Mouse Anti-Human IL‑27/IL‑35 EBI3 Subunit Alexa Fluor® 488‑conjugated Monoclonal Antibody (Catalog # IC6456G) or isotype control antibody (Catalog # IC002G). View our protocol for Staining Intracellular Molecules.|
is a heterodimeric cytokine comprised of the IL-12 family that is composed of two chains, EBV-induced gene 3( EBI3), a 33 kDa glycosylated protein homologous to the p40 subunit of IL-12, and p28, a 28 kDa peptide analogous to the p35 subunit of IL-12. IL-27 has been found to be expressed by activated monocytes and monocyte-derived dendritic cells, astrocytes, adipocytes, CD4+ T cells, and macrophages. In human tissues, EBI3 and p28 coexpression has been observed in macrophages or macrophage-derived cells, and in endothelial cells. The IL-27 receptor complex comprises two chains, IL-27R (also called WSX-1 or TCCR) and gp130. IL-27 is the only known ligand for IL-27R, whereas gp130 is a receptor component of several cytokines of the IL-6 family. Functionally, IL-27 appears to be immunosuppressive. On T cells in mice, it induces IL-10 secretion while suppressing IL-17A and GM-CSF production. But the relationship between IL-27 and IL-17A may be complex. After IL-17A induction, IL-27 secretion occurs in unstimulated cells, suggesting IL-27 downregulates bystander activation in inflammatory conditions. EBI3 also complexes with p35 (IL-12) to form IL-35. The mature EBI3 sequences for human and mouse share 62% amino acid sequence identity.