烟草烟雾诱发哮喘异质性，呈难治性特质，其具体发病机制及防控措施尚未阐明。申报人前期证实CpG-ODN抑制TSLP驱动树突细胞（DC）介导过敏性哮喘的Th2反应。预试验示烟雾哮喘呈Th2/Th17极化，而CpG-ODN通过TSLP-DC减轻Th17反应，并调节促炎因子IL-33和抑炎因子IL-35。据此我们推测CpG-ODN通过调控IL-33/ST2和IL-35/EBI3信号，从上游抑制TSLP-DC途径，从而在下游下调烟雾哮喘Th17反应。在前期研究的基础上拟利用CRISPR/Cas9条件性基因敲除鼠、烟草烟雾提取物/CpG-ODN预处理DC过继转移、及IL-33、EBI3腺病毒载体转染DC构建模型，并结合吸烟哮喘患者体外实验，阐明IL-33/ST2和IL-35/EBI3在烟雾哮喘Th17反应的关键作用及CpG-ODN的调控机制。项目将为寻找烟雾哮喘干预靶点和治疗新策略提供理论及实验依据。

The heterogeneity of asthma induced by cigarette smoking (CS) exposure is an important factor in refractory asthma. Therefore, it is of great importance in both theory and in clinical practice to explore molecular mechanisms and control measures of CS exposure. Our previous studies have revealed that CpG-ODN inhibits Th2-type response in allergic asthma mediated by blocking thymic stromal lymphopoietin (TSLP)-dendritic cell (DC) pathway. Furthermore, our preliminary experiments showed that CS exposure results in a phenotype with mixed inflammatory cells, airway inflammation, airway remodeling, and Th2/Th17 skewing, while CpG-ODN dampens Th2/Th17 polarization via TSLP-DC pathway, simultaneously reduces the secretion of proinflammatory cytokine IL-33, and also facilitates the secretion of anti-inflammatory cytokine IL-35. However, the question that, whether or not CpG-ODN regulates IL-33/ST2 and IL-35/EBI3 in the upstream, which subsequently contributes to alleviate inhibiting CS-exposed asthma Th17-oriented response mediated by TSLP-DC pathway in the downstream, is still unclear. According to our previous observations, it seems to be logical to assume that CpG-ODN inhibits Th2/Th17 polarization of CS exposure asthma mediated by TSLP-DC pathway by regulating IL-33/ST2 and IL-35/EBI3 signaling pathways. In order to prove this hypothesis, a murine model of CS-exposed mice was established using CRISPR/Cas9 mediated lung-specific knockout of IL-33 and EBI3 genes, adoptive transfer of OVA pulsed DCs treated by CS extract and/or CpG-ODN, or IL-33, EBI3 gene-modulated adenovirus infection of DC, combined with in vitro clinical study of asthmatic patients with current smoker, aiming to elucidate the key role of IL-33/ST2 and IL-35/EBI3 in the development of Th17 response and the regulatory mechanisms of CpG-ODN underlying CS exposure asthma. The present research aimed to reveal a promising target for CS exposure asthma therapy and provide a theoretical and reliable experimental basis for developing new treatment strategies.