Increased expression of upstream TH2-cytokines in a mouse model of viral-induced asthma exacerbation.
Increased expression of upstream TH2-cytokines in a mouse model of viral-induced asthma exacerbation.
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DOI：
10.1186/s12967-016-0808-x
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发表时间：
2016-02-16
影响因子：
7.4
通讯作者：
Uller L
中科院分区：
文献类型：
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作者：
Mahmutovic Persson I;Akbarshahi H;Menzel M;Brandelius A;Uller L
呼吸道病毒感染引起的哮喘恶化是一种严重的疾病，需要新的治疗方法。为此目的，哮喘恶化的动物模型是有根据的。我们已经证明dsRNA挑战或鼻病毒感染对卵清蛋白（OVA）诱导的过敏性哮喘小鼠产生加重作用。然而，屋尘螨（HDM）是一种与人类哮喘相关的过敏原，而不是OVA。因此，我们假设hdm诱导的实验性哮喘小鼠的dsRNA挑战将产生哮喘加重的重要翻译特征。用HDM或生理盐水局部刺激小鼠气道，每周3次，连续3周建立实验性哮喘。然后连续3天每天给予局部dsRNA刺激以诱导加重。分析支气管肺泡灌洗液（BALF）中炎症细胞、总蛋白、坏死标志物LDH和报警蛋白ATP的含量。分析肺匀浆中TNF-α、CCL2、CCL5、IL-1β、IL-33、胸腺基质淋巴生成素（TSLP）、IL-25以及模式识别受体（PRRs） rig -1、MDA5和TLR3的mRNA表达情况（RT-qPCR）。ELISA法检测肺组织IL-33， western blot法检测PRRs。免疫组化显示IL-33在肺中的分布。单独攻毒HDM引起BALF总蛋白、嗜酸性粒细胞、淋巴细胞和中性粒细胞持续升高，肺组织TSLP、IL-33和TNF-α表达短暂升高。dsrna诱导的急性加重明显且剂量依赖性地夸大了这些作用。此外，LDH和ATP的BALF水平以及肺组织CCL2、CCL5、IL-1β、IL-25和PRRs的表达均在加重时升高。肺中IL-33蛋白水平在HDM作用下短暂升高，加重时进一步升高。我们展示了hdm诱导的实验性哮喘的几个新方面和dsRNA的加重作用。BALF中的一般炎症参数，如渗出蛋白、混合粒细胞、LDH和ATP，在目前的发作中与人类哮喘发作时一样升高。我们认为，这种涉及dsRNA挑战的哮喘加重模型具有翻译价值，并且表明它可能特别适合于体内研究，涉及对加重诱导的主要上游th2细胞因子表达的药理作用；IL-33， TSLP和IL-25，以及PRRs。本文的在线版本（doi:10.1186/s12967-016-0808-x）包含补充材料，可供授权用户使用。
Exacerbations of asthma caused by respiratory viral infections are serious conditions in need of novel treatment. To this end animal models of asthma exacerbations are warranted. We have shown that dsRNA challenges or rhinoviral infection produce exacerbation effects in mice with ovalbumin (OVA)-induced allergic asthma. However, house dust mite (HDM) is a more human asthma-relevant allergen than OVA. We thus hypothesised that dsRNA challenges in mice with HDM-induced experimental asthma would produce important translational features of asthma exacerbations. Mouse airways were challenged locally with HDM or saline three times a week for three weeks to establish experimental asthma. Then daily local dsRNA challenges were given for three consecutive days to induce exacerbation. Bronchoalveolar lavage fluid (BALF) was analysed for inflammatory cells, total protein, the necrosis marker LDH and the alarmin ATP. Lung homogenates were analysed for mRNA expression (RT-qPCR) of TNF-α, CCL2, CCL5, IL-1β, IL-33, thymic stromal lymphopoietin (TSLP), and IL-25 as well as pattern recognition receptors (PRRs) RIG-I, MDA5 and TLR3. Lung tissue IL-33 was analysed with ELISA and PRRs were quantified by western blot. Immunohistochemistry indicated lung distribution of IL-33. HDM challenge alone caused sustained increase in BALF total protein, eosinophils, lymphocytes and neutrophils, and transient increase in lung tissue expression of TSLP, IL-33 and TNF-α. dsRNA-induced exacerbation markedly and dose-dependently exaggerated these effects. Further, BALF levels of LDH and ATP, and lung tissue expression of CCL2, CCL5, IL-1β, IL-25 and PRRs were increased exclusively at the exacerbations. Lung protein levels of IL-33 were transiently increased by HDM and further increased at exacerbation. We demonstrate several novel aspects of HDM-induced experimental asthma and added exacerbation effects of dsRNA. General inflammatory parameters in BALF such as exuded proteins, mixed granulocytes, LDH and ATP were increased at the present exacerbations as they are in human asthma exacerbations. We suggest that this model of asthma exacerbation involving dsRNA challenges given to mice with established HDM-induced asthma has translational value and suggest that it may be particularly suited for in vivo studies involving pharmacological effects on exacerbation-induced expression of major upstream TH2-cytokines; IL-33, TSLP and IL-25, as well as PRRs. The online version of this article (doi:10.1186/s12967-016-0808-x) contains supplementary material, which is available to authorized users.