IL-9-Mediated Induction of Eotaxin1/CCL11 in Human Airway Smooth Muscle Cells

Abstract

Recent work has shown the potential importance of IL-9 in allergic diseases. The development of transgenic mice overexpressing IL-9 has suggested a key role for this cytokine in the development of the asthmatic phenotype including airway eosinophilia. In this study, we evaluated the expression of the IL-9R and the effects of IL-9 on human ASM cells by examining the release of Th2-associated chemokines (eotaxin1/CCL11 and thymus- and activation-regulated chemokine (TARC)/CCL17). IL-9R α-chain mRNA and surface expression were detected in cultured human airway smooth muscle (ASM) cells. In addition, primary cultured ASM cells, as well as bronchial smooth muscle cells within biopsies of asthmatics and not control subjects, revealed IL-9R protein expression. IL-9 stimulation of human ASM cells resulted in release of eotaxin1/CCL11, but had no effect on the release of TARC/CCL17, in time- and dose-dependent manner. Moreover, in vitro chemotaxis assay demonstrated that conditioned medium from IL-9-stimulated ASM cells attracted human eosinophils. Neutralizing Abs to IL-9, but not to IL-4 or IL-13, reduced significantly IL-9-induced production of eotaxin1/CCL11 from ASM cells. Interestingly, real-time RT-PCR showed that IL-9 up-regulated eotaxin1/CCL11 mRNA expression, but had no effect on TARC/CCL17. Treatment with Act D abrogates IL-9-induced eotaxin1/CCL11 mRNA and protein release by ASM cells. Finally, transfection study using eotaxin1/CCL11 promoter luciferase construct confirmed that IL-9 induced eotaxin1/CCL11 at the transcriptional level. Taken together, these data provide new evidence demonstrating that IL-9-dependent activation of ASM cells contributes to eosinophilic inflammation observed in asthma.

Thymic Stromal Lymphopoietin Receptor-Mediated IL-6 and CC/CXC Chemokines Expression in Human Airway Smooth Muscle Cells: Role of MAPKs (ERK1/2, p38, and JNK) and STAT3 Pathways

“…Immunoreactive IL-6 was quantified by ELISA using matched Abs from BioLegend (San Diego, CA), according to basic laboratory protocols. Immunoreactive IL-8/CXCL8 and eotaxin-1/CCL11 were measured by ELISA as described previously (25,27). The sensitivity limit for both IL-8/CXCL8 and IL-6 was 10 and 7.8 pg/ml for eotaxin-1/CCL11.…”

“…pp , 0.05; ppp , 0.01 (n = 3) compared with wild-type luciferase activity in unstimulated cells. TSLP induces the promoter activation of IL-8/CXCL8, eotaxin-1/CCL11, and IL-6 genes To further investigate the mechanisms by which TSLP mediates cytokine/chemokine expression, HASM cells were transiently transfected with wild-type luciferase reporter constructs for IL-8/ CXCL8 (25), eotaxin-1/CCL11 (27), and IL-6 (30) carrying proximal promoter regions of respective cytokine/chemokine genes. Primary HASM cells transfected with IL-8/CXCL8, eotaxin-1/CCL11, or IL-6 promoter construct showed a significant increase in luciferase activity (p , 0.05; n = 3) in response to TSLP (0.1 ng/ml) stimulation (mean value of fold increase compared with baseline: 1.24 6 0.19, 1.18 6 0.13, and 1.22 6 0.06, respectively) (Fig.…”

IL‐9 promotes the survival and function of human melanoma‐infiltrating CD4+CD8+ double‐positive T cells

“…In turn, Lu et al described another antitumor effect of IL-9 on melanoma lung metastasis, which through the induction of local CCL20 expression in the tumor microenvironment, led to an enhanced recruitment of dendritic cells and cytotoxic CD8 + T cells at the tumor site [58]. The ability of IL-9 to induce the secretion of chemotactic factors was also largely described in airway inflammatory pathologies [59]. Therefore, we can postulate that a similar process occurs in the lymph node metastasis environment and could contribute to the increase frequency of intramelanoma DP T cells.…”

Emerging Role of Interleukins in Cancer Treatment

“…These mechanisms include activation of mast cells. IL-9 also induces the release of Th2-associated chemokines in cultured human airway smooth muscle cells [113] and enhanced the stem cell factor-dependent growth of human mast cell progenitors, particularly those of children with asthma.…”

“…MEDI-528 is a humanized immunoglobulin G1 monoclonal antibody that binds to IL-9, and hence reduces the activity of a variety of cell types implicated in asthma pathogenesis [113,114]. Despite available phase I and II clinical trials on safety and efficacy of this anti-IL-9 antibody in severe uncontrolled asthma, its clinical application and benefit in hematologic malignancies was not identified.…”