The role of ovarian hormones on allergen-mediatedd innate immune airway responses

 描述(申请人提供)：哮喘患病率在男孩中比女孩更高，但在青春期前后哮喘患病率有所变化，中年女性患哮喘的可能性大约是男性的两倍。这暗示了性激素在哮喘发病机制中的作用；然而，其机制仍不清楚。第二组先天淋巴样细胞(ILC2)在哮喘相关的过敏性呼吸道炎症的初始阶段起重要作用。ILC2是由IL-33、TSLP和IL-25激活的，IL-33、TSLP和IL-25是对包括交链孢属在内的呼吸道变应原的上调。在刺激下，ILC2可增加转录因子GATA3和RoRα的表达，并产生IL-5和IL-13。IL-5和IL-13增加嗜酸性粒细胞的渗透、气道高反应性(AHR)和粘液产生，这些都是哮喘的特征。我们的初步数据显示，与雄性小鼠的ILC2相比，雌性小鼠的ILC2显著增加了IL-5和IL-13的蛋白表达。因此，我们推测卵巢激素17β-雌二醇(17-β-E_2)和孕酮(P_4)增加了ILC2诱导的气道炎症。在目标1中，我们将确定性激素促进ILC2产生IL-5和IL-13的机制。假手术雌雄小鼠、去卵巢雌性小鼠和去睾丸雄性小鼠将被给予含有赋形剂、17β-E_2、P4和/或睾酮的微丸。从这些小鼠的骨髓和肺中获取ILCs，并在体外用IL-33刺激。IL-5和IL-13的蛋白表达，RoRα和GATA3mRNA的表达，以及IL-33受体的一种成分ST2的表面表达将在ILC2中进行测定。在目标2中，我们将确定卵巢激素在链格孢霉提取物(Alt Ex)诱导的先天免疫介导的呼吸道炎症中的作用。WT BALB/c雌性、雄性、去卵巢和去的小鼠将被Alt Ex激发4天以启动先天免疫反应。然后我们将测定ILC2细胞因子的表达、呼吸道炎症、AHR和粘液产生。这项建议将勾勒出性激素调节ILC2的机制(S)，并可能确定哮喘患者，特别是女性哮喘患者的潜在治疗靶点。

 DESCRIPTION (provided by applicant): Asthma prevalence is greater in boys than girls, but around puberty there is a shift in asthma prevalence, and at mid-life women are about two times more likely than men to have asthma. This suggests a role for sex hormones in asthma pathogenesis; however the mechanisms remain unknown. Group 2 innate lymphoid cells (ILC2) are important in driving the initial phase of allergic airway inflammation that is associated with asthma. ILC2 are activated by IL-33, TSLP, and IL-25 which are upregulated in response to airway allergens, including Alternaria alternata. Upon stimulation, ILC2 have increased expression of the transcription factors GATA3 and RORα and produce IL-5 and IL-13. IL-5 and IL-13 increase the infiltration of eosinophils, airway hyperresponsiveness (AHR), and mucus production, all hallmarks of asthma. Our preliminary data showed IL- 33-stimulated ILC2 from female mice had significantly increased IL-5 and IL-13 protein expression compared to ILC2 from male mice. Therefore, we hypothesize that the ovarian hormones, 17beta-estradiol (17β-E2) and progesterone (P4), increase ILC2-induced airway inflammation. In Aim 1 we will determine the mechanisms by which sex hormones increase IL-5 and IL-13 production from ILC2. Sham-operated female and male mice, ovariectomized female mice, and orchidectomized male mice will be administered pellets containing vehicle, 17β-E2, P4, and/or testosterone. ILCs will be harvested from the bone marrow and lungs of these mice and stimulated with IL-33 ex vivo. IL-5 and IL-13 protein expression, RORα and GATA3 mRNA expression, and surface expression of ST2, a component of the IL-33 receptor, will then be determined in ILC2. In Aim 2, we will determine the role of ovarian hormones on Alternaria extract (Alt Ex)-induced innate immune-mediated airway inflammation. WT BALB/c female, male, ovariectomized, and orchidectomized mice will be challenged with Alt Ex for 4 days to initiate an innate immune response. We will then determine ILC2 cytokine expression, airway inflammation, AHR, and mucus production. This proposal will delineate a mechanism(s) by which sex hormones regulate ILC2, and it may identify potential therapeutic targets for patients, in particular women, with asthma.