|Cell Proliferation Induced by IL‑9 and Neutralization by Rat IL‑9 Antibody. Recombinant Rat IL‑9 (Catalog # 4134-RL) stimulates proliferation in the TS1 mouse helper T cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat IL‑9 (10 ng/mL) is neutralized (green line) by increasing concentrations of Sheep Anti-Rat IL‑9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4134). The ND50 is typically 1.5‑7.5 µg/mL.|
Intracellular Staining by Flow Cytometry
|Detection of IL‑9 in Rat Splenocytes by Flow Cytometry. Rat splenocytes were treated for 24 hours with 50 ng/mL PMA and 500 ng/mL Ca2+ ionomycin then stained with Sheep Anti-Rat IL‑9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4134, filled histogram) or control antibody (Catalog # 5‑001‑A, open histogram), followed by NorthernLights™ 637-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # NL011). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.|
Rat interleukin-9 (IL-9; also T-cell growth factor P40 and MEA) is a 20‑30 kDa secreted glycoprotein that belongs to the IL-7/IL-9 family of hematopoietic cytokines. It is best considered an atypical Th2-type cytokine (1‑4). Rat IL-9 was initially isolated from the 4437A rat thymoma cell line (5). The IL-9 precursor is 144 amino acids (aa) in length and contains an 18 aa signal sequence plus a 126 aa mature segment (5). An extended, 219 aa alternate precursor form is reported in GenBank (Accession # XP_001068265). It shows a 113 aa substitution for the first 38 aa of the standard form. Its significance is unknown. Mature rat IL-9 is 75%, 74% and 58% aa identical to cotton rat, mouse and human IL-9, respectively (6‑9). There is variability in cross-species reactivity. Canine and mouse IL-9 are purportedly active on human cells (9‑11), while human IL-9 is inactive on mouse cells (9). Cells reported to express IL-9 include activated CD4+ T cells and eosinophils (12, 13). The receptor for rat IL-9 is composed of the 62 kDa rat IL-9R alpha chain and the 64 kDa rat common cytokine receptor gamma -chain (5, 14, 15). IL-9 is suggested to play a significant, but likely supporting role in allergy and asthma. Its synthesis is induced upon exposure to allergen (10, 13). Following release, it is known to upregulate IgE production by IL‑4‑stimulated B cells (16). It also stimulates mucin secretion and IL-13 production through a direct action on respiratory epithelium (10, 17). In the bone marrow, it promotes eosinophilia, likely through IL-5 induction (18, 19). And once produced, eosinophils are chemoattracted to the bronchi through the IL-9-induced secretion of eotaxin-1 by airway smooth muscle cells (20).
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