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Multicolor flow cytometric analysis of IL-17 expression in stimulated human lymphocytes. An enriched preparation of human peripheral blood CD4+ T cells was stimulated with immobilized Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329; 10 μg/ml for plate coating) and soluble Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725; 2 μg/ml) antibodies and Recombinant Human IL-2 (Cat. No. 554603; 10 ng/ml) and Human IL-4 (Cat. No. 554605; 25 ng/ml) proteins for 2 days. The stimulated cells were washed and cultured in medium containing IL-2 and IL-4 for another 3 days. Finally, the cells were harvested and restimulated for 5 hr with Phorbol 12-Myristate 13-Acetate (PMA; Sigma, Cat. No. P-8139; 50 ng/ml) and Ionomycin (Sigma, Cat. No. I-0634; 1 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested and washed with Stain Buffer (FBS) (Cat. No. 554656) and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BUV395 Mouse Anti-Human CD4 (Cat No. 564724), FITC Mouse Anti-Human CD45RO (Cat. No. 555492; Left Plot), BD Horizon™ BV421 Mouse Anti-Human IL-4 (Cat. No. 564110; Middle Plot Plot), and PE Mouse Anti-Human IFN-γ (Cat. No. 559326; Right Plot Plot) antibodies and with either BD Horizon™ R718 Rat IgG1 κ Isotype Control (Cat. No. 566928; Data Not Shown) or BD Horizon™ R718 Mouse Anti-Human IL-17 antibody (Cat. No. 567094/567247) using BD Biosciences Intracellular Cytokine Staining protocol. Pseudocolor density plots showing the correlated expression of IL-17 versus CD45RO, IL-4, or IFN-γ were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact stimulated lymphocytes as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
BD Horizon™ R718 Mouse Anti-Human IL-17A
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Human IL-17A, also known as IL-17, is a proinflammatory cytokine that is encoded by the IL17A gene in chromosome 6. IL-17A is produced as a disulfide-linked homodimer comprised of two mature 136-amino acid polypeptides. It is a member of the IL-17 family of structurally related cytokines, designated IL-17A through IL-17F. Activated memory T cells, especially Th17 cells (specialized IL-17A-producing CD4+ T cells distinct from Th1 and Th2 cells) produce IL-17 and provide protective immunity against pathogens. Activated CD8+ T cells, γδT cells, NK cells and neutrophils can also be activated to produce IL-17A. IL-17A binds to and exerts its biological activity through IL-17 receptors (IL-17R) that are expressed by a variety of target cells including fibroblasts, epithelial and endothelial cells, monocytes/macrophages and mast cells. The ubiquitous IL-17R expression pattern may explain the broad tissue responsiveness to IL-17. IL-17 induces stromal cells to secrete cytokines and chemokines involved in inflammatory and hematopoietic processes. For example, IL-17 induces fibroblasts to produce IL-6, IL-8, G-CSF and express increased surface ICAM-1. The SCPL1362 antibody reacts with human IL-17A.
Development References (6)
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Fossiez F, Djossou O, Chomarat P, et al. T cell interleukin-17 induces stromal cells to produce proinflammatory and hematopoietic cytokines. J Exp Med. 1996; 183(6):2593-2603. (Biology). View Reference
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Korn T, Oukka M, Kuchroo V, Bettelli E. Th17 cells: effector T cells with inflammatory properties. Semin Immunol. 2007; 19(6):362-371. (Biology). View Reference
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Moseley TA, Haudenschild DR, Rose L, Reddi AH. Interleukin-17 family and IL-17 receptors. Cytokine Growth Factor Rev. 2003; 14(2):155-174. (Biology). View Reference
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Weaver CT, Hatton RD, Mangan PR, Harrington LE. IL-17 family cytokines and the expanding diversity of effector T cell lineages. Annu Rev Immunol. 2007; 25:821-852. (Biology). View Reference
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Yao Z, Painter SL, Fanslow WC, et al. Human IL-17: a novel cytokine derived from T cells. J Immunol. 1995; 155(12):5483-5486. (Biology). View Reference
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Yao Z, Spriggs MK, Derry JM, et al. Molecular characterization of the human interleukin (IL)-17 receptor. Cytokine. 1997; 9(11):794-800. (Biology). View Reference
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