Gngt2 通过 ATG5-NF-κB 信号轴促进中性粒细胞型哮喘中炎症性树突状细胞分化定型
2026/06/30
摘要
背景：低2型免疫（T2）中性粒细胞型哮喘属于重症哮喘内型，核心特征为激素抵抗，且气道持续性炎症由树突状细胞（DC）介导的辅助性T细胞17（Th17）应答所驱动。尽管免疫原性树突状细胞分化定型是该病理进程的关键环节，但将炎症信号通路与树突状细胞功能极化相耦联的胞内调控机制尚不明确。
目的：本研究旨在探究G蛋白γ转导素亚基2（Gngt2）是否可调控树突状细胞中自噬相关蛋白依赖性炎症分化进程，进而参与 Th17 介导的中性粒细胞性气道炎症。
方法：本研究通过构建 BALB/c 小鼠卵清蛋白（OVA）诱导嗜酸粒细胞型哮喘模型、卵清蛋白联合脂多糖（LPS）诱导中性粒细胞型哮喘模型，对比两种哮喘内型特异性炎症应答差异；经气道滴注慢病毒短发夹RNA实现体内Gngt2基因沉默。体外培养骨髓来源树突状细胞（BMDC），分别给予卵清蛋白、卵清蛋白联合脂多糖刺激，并设置Gngt2敲低、ATG5敲低、ATG5过表达干预组；部分实验采用NF-κB核转位抑制剂JSH-23验证信号通路依赖性。将骨髓来源树突状细胞与初始CD4+T细胞共培养，检测Th17细胞分化水平；结合组织学、免疫学及分子生物学检测，评估气道炎症程度、细胞因子分泌、自噬相关蛋白表达及NF-κB活化水平。
结果：中性粒细胞型哮喘小鼠肺组织树突状细胞中Gngt2呈选择性高表达，伴随自噬相关蛋白表型紊乱：ATG5 蛋白表达下调、微管相关蛋白1轻链 3β（LC3B）表达降低、LC3B-II/I 比值下降。体内沉默 Gngt2 可显著减轻中性粒细胞性气道炎症，降低支气管肺泡灌洗液（BALF）中白细胞介素- 6（IL-6）、白细胞介素- 23（IL-23）、白细胞介素- 17（IL-17）浓度，对白介素- 4（IL-4）无显著影响。在卵清蛋白联合脂多糖刺激的骨髓来源树突状细胞中，敲低Gngt2可恢复ATG5/LC3B自噬相关标志物表达，抑制NF-κB p65磷酸化，减少 IL-6、IL-23分泌，并阻断树突状细胞介导的 Th17 细胞分化。回复实验进一步证实，ATG5 位于Gngt2下游，调控NF-κB 活化与炎症因子释放，进而调控Th17细胞极化。
结论：本研究首次发现树突状细胞内源Gngt2-ATG5-NF-κB信号轴，该通路将G蛋白偶联受体（GPCR）信号与自噬相关蛋白异常重塑的免疫调控相衔接，参与中性粒细胞型哮喘发病。Gngt2通过抑制ATG5依赖性信号通路，正向促进NF-κB活化，加重Th17介导的中性粒细胞性气道炎症。该通路阐明了低T2型哮喘中树突状细胞介导免疫紊乱的分子机制，提示 Gngt2可作为中性粒细胞型哮喘潜在治疗靶点。
Gngt2 promotes inflammatory dendritic cell programming through an ATG5-NF-κB signaling axis in neutrophilic asthma.
Ji X, He SD, Yang L, You X, Wen T, Chen Z, Zhou Y.
Abstract
BACKGROUND:Type 2 (T2)-low neutrophilic asthma represents a severe asthma endotype characterized by corticosteroid resistance and persistent airway inflammation driven by dendritic cells (DCs)-mediated T helper (Th)17 responses. Although immunogenic DC programming plays a central role in this process, the intracellular mechanisms that couple inflammatory signaling to DC functional polarization remain poorly understood.
OBJECTIVE:This study investigated whether the G protein subunit gamma transducin 2 (Gngt2) regulates autophagy related protein-dependent inflammatory programming in DCs and thereby contributes to Th17-driven neutrophilic airway inflammation.
METHODS:Ovalbumin (OVA)-induced eosinophilic asthma and OVA/lipopolysaccharide (LPS)-induced neutrophilic asthma models were established in BALB/c mice to compare endotype-specific inflammatory responses. Gngt2 was silenced in vivo by intratracheal delivery of lentiviral short hairpin RNA. In vitro, bone marrow-derived dendritic cells (BMDCs) were stimulated with OVA or OVA/LPS and then manipulated by Gngt2 knockdown, ATG5 knockdown, or ATG5 overexpression. In selected experiments, the NF-κB nuclear translocation inhibitor JSH-23 was used to assess pathway dependence. BMDCs were co-cultured with naïve CD4 + T cells to evaluate Th17 differentiation. Airway inflammation, cytokine production, autophagy-related protein expression, and NF-κB activation were assessed by histological, immunological, and molecular analyses.
RESULTS:Gngt2 expression was selectively upregulated in lung DCs from mice with neutrophilic asthma and was accompanied by a phenotype with altered autophagy related protein, as indicated by reduced ATG5 abundance, decreased microtubule-associated protein 1 light chain 3 beta (LC3B) expression, and a lower LC3B-II/I ratio. In vivo silencing of Gngt2 markedly attenuated neutrophilic airway inflammation and decreased bronchoalveolar lavage fluid (BALF) levels of interleukin (IL)-6, IL-23, and IL-17, without significantly affecting IL-4. In OVA/LPS-stimulated BMDCs, Gngt2 knockdown restored ATG5/LC3B-associated autophagy-related markers, reduced NF-κB p65 phosphorylation, suppressed IL-6 and IL-23 production, and impaired DC-mediated Th17 differentiation. Rescue experiments further demonstrated that ATG5 acted downstream of Gngt2 to regulate NF-κB activation and inflammatory cytokine production, thereby controlling Th17 polarization.
CONCLUSION:Our findings identify a previously unrecognized DC-intrinsic Gngt2-ATG5-NF-κB signaling axis that links G protein-coupled receptor (GPCR)- signaling with immune regulation of altered autophagy related protein in neutrophilic asthma. By suppressing ATG5-mediated signaling, Gngt2 promotes NF-κB activation and contributes to Th17-driven neutrophilic airway inflammation. This pathway provides mechanistic insight into DC-driven immune dysregulation in type 2-low asthma and suggests Gngt2 as a potential therapeutic target for neutrophilic asthma.
背景：低2型免疫（T2）中性粒细胞型哮喘属于重症哮喘内型，核心特征为激素抵抗，且气道持续性炎症由树突状细胞（DC）介导的辅助性T细胞17（Th17）应答所驱动。尽管免疫原性树突状细胞分化定型是该病理进程的关键环节，但将炎症信号通路与树突状细胞功能极化相耦联的胞内调控机制尚不明确。
目的：本研究旨在探究G蛋白γ转导素亚基2（Gngt2）是否可调控树突状细胞中自噬相关蛋白依赖性炎症分化进程，进而参与 Th17 介导的中性粒细胞性气道炎症。
方法：本研究通过构建 BALB/c 小鼠卵清蛋白（OVA）诱导嗜酸粒细胞型哮喘模型、卵清蛋白联合脂多糖（LPS）诱导中性粒细胞型哮喘模型，对比两种哮喘内型特异性炎症应答差异；经气道滴注慢病毒短发夹RNA实现体内Gngt2基因沉默。体外培养骨髓来源树突状细胞（BMDC），分别给予卵清蛋白、卵清蛋白联合脂多糖刺激，并设置Gngt2敲低、ATG5敲低、ATG5过表达干预组；部分实验采用NF-κB核转位抑制剂JSH-23验证信号通路依赖性。将骨髓来源树突状细胞与初始CD4+T细胞共培养，检测Th17细胞分化水平；结合组织学、免疫学及分子生物学检测，评估气道炎症程度、细胞因子分泌、自噬相关蛋白表达及NF-κB活化水平。
结果：中性粒细胞型哮喘小鼠肺组织树突状细胞中Gngt2呈选择性高表达，伴随自噬相关蛋白表型紊乱：ATG5 蛋白表达下调、微管相关蛋白1轻链 3β（LC3B）表达降低、LC3B-II/I 比值下降。体内沉默 Gngt2 可显著减轻中性粒细胞性气道炎症，降低支气管肺泡灌洗液（BALF）中白细胞介素- 6（IL-6）、白细胞介素- 23（IL-23）、白细胞介素- 17（IL-17）浓度，对白介素- 4（IL-4）无显著影响。在卵清蛋白联合脂多糖刺激的骨髓来源树突状细胞中，敲低Gngt2可恢复ATG5/LC3B自噬相关标志物表达，抑制NF-κB p65磷酸化，减少 IL-6、IL-23分泌，并阻断树突状细胞介导的 Th17 细胞分化。回复实验进一步证实，ATG5 位于Gngt2下游，调控NF-κB 活化与炎症因子释放，进而调控Th17细胞极化。
结论：本研究首次发现树突状细胞内源Gngt2-ATG5-NF-κB信号轴，该通路将G蛋白偶联受体（GPCR）信号与自噬相关蛋白异常重塑的免疫调控相衔接，参与中性粒细胞型哮喘发病。Gngt2通过抑制ATG5依赖性信号通路，正向促进NF-κB活化，加重Th17介导的中性粒细胞性气道炎症。该通路阐明了低T2型哮喘中树突状细胞介导免疫紊乱的分子机制，提示 Gngt2可作为中性粒细胞型哮喘潜在治疗靶点。
（中日友好医院呼吸与危重症医学科 张婧媛 摘译 林江涛 审校）
（Inflamm Res. 2026 Jun 2;75(1):126. doi: 10.1007/s00011-026-02276-9. PMID: 42228185.）
（Inflamm Res. 2026 Jun 2;75(1):126. doi: 10.1007/s00011-026-02276-9. PMID: 42228185.）
Gngt2 promotes inflammatory dendritic cell programming through an ATG5-NF-κB signaling axis in neutrophilic asthma.
Ji X, He SD, Yang L, You X, Wen T, Chen Z, Zhou Y.
Abstract
BACKGROUND:Type 2 (T2)-low neutrophilic asthma represents a severe asthma endotype characterized by corticosteroid resistance and persistent airway inflammation driven by dendritic cells (DCs)-mediated T helper (Th)17 responses. Although immunogenic DC programming plays a central role in this process, the intracellular mechanisms that couple inflammatory signaling to DC functional polarization remain poorly understood.
OBJECTIVE:This study investigated whether the G protein subunit gamma transducin 2 (Gngt2) regulates autophagy related protein-dependent inflammatory programming in DCs and thereby contributes to Th17-driven neutrophilic airway inflammation.
METHODS:Ovalbumin (OVA)-induced eosinophilic asthma and OVA/lipopolysaccharide (LPS)-induced neutrophilic asthma models were established in BALB/c mice to compare endotype-specific inflammatory responses. Gngt2 was silenced in vivo by intratracheal delivery of lentiviral short hairpin RNA. In vitro, bone marrow-derived dendritic cells (BMDCs) were stimulated with OVA or OVA/LPS and then manipulated by Gngt2 knockdown, ATG5 knockdown, or ATG5 overexpression. In selected experiments, the NF-κB nuclear translocation inhibitor JSH-23 was used to assess pathway dependence. BMDCs were co-cultured with naïve CD4 + T cells to evaluate Th17 differentiation. Airway inflammation, cytokine production, autophagy-related protein expression, and NF-κB activation were assessed by histological, immunological, and molecular analyses.
RESULTS:Gngt2 expression was selectively upregulated in lung DCs from mice with neutrophilic asthma and was accompanied by a phenotype with altered autophagy related protein, as indicated by reduced ATG5 abundance, decreased microtubule-associated protein 1 light chain 3 beta (LC3B) expression, and a lower LC3B-II/I ratio. In vivo silencing of Gngt2 markedly attenuated neutrophilic airway inflammation and decreased bronchoalveolar lavage fluid (BALF) levels of interleukin (IL)-6, IL-23, and IL-17, without significantly affecting IL-4. In OVA/LPS-stimulated BMDCs, Gngt2 knockdown restored ATG5/LC3B-associated autophagy-related markers, reduced NF-κB p65 phosphorylation, suppressed IL-6 and IL-23 production, and impaired DC-mediated Th17 differentiation. Rescue experiments further demonstrated that ATG5 acted downstream of Gngt2 to regulate NF-κB activation and inflammatory cytokine production, thereby controlling Th17 polarization.
CONCLUSION:Our findings identify a previously unrecognized DC-intrinsic Gngt2-ATG5-NF-κB signaling axis that links G protein-coupled receptor (GPCR)- signaling with immune regulation of altered autophagy related protein in neutrophilic asthma. By suppressing ATG5-mediated signaling, Gngt2 promotes NF-κB activation and contributes to Th17-driven neutrophilic airway inflammation. This pathway provides mechanistic insight into DC-driven immune dysregulation in type 2-low asthma and suggests Gngt2 as a potential therapeutic target for neutrophilic asthma.
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