高迁移率族蛋白B1通过调节博来霉素诱导的肺纤维化中的自噬来调控树突状细胞的活化和CD4+ T细胞反应。

HMGB1 regulates the activation of dendritic cells and CD4+ T cell responses through the modulation of autophagy in bleomycin-induced pulmonary fibrosis.

作者信息

Liu Xiuhua, Song Xinghui, Li Guangting, Zhang Yuping, Liu Nina, Tang Kaijiang, Du Hongyan, Jie Ligang

机构信息

Department of rheumatology and clinical immunology, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510000, China; Department of Rheumatism and Immunology, The Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou, Guangxi 545007, China.

Department of Rheumatism and Immunology, The Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou, Guangxi 545007, China.

出版信息

Immunobiology. 2025 May;230(3):152906. doi: 10.1016/j.imbio.2025.152906. Epub 2025 Apr 23.

DOI:10.1016/j.imbio.2025.152906

PMID:40311346

Abstract

BACKGROUND

The role of HMGB1 in inflammation and autophagy has garnered increasing attention; however, its impact on the activation of dendritic cells (DCs) and autophagy remains unclear. This study aims to explore the effects of HMGB1 on DC activation, autophagy, and its influence on CD4+ T cell responses in a bleomycin-induced pulmonary fibrosis (PF) mouse model.

METHODS

Thirty mice were randomly divided into control and model groups. The model group was established by intratracheal injection of bleomycin to induce PF. Flow cytometry was used to detect DC surface markers, and western blot was employed to assess the expression of autophagy-related protein LC3. Lung DCs and peripheral blood CD14+ monocytes were sorted using magnetic beads and differentiated into M0-DCs, which were then subjected to HMGB1 stimulation experiments to assess activation and cytokine secretion. HMGB1-stimulated or untreated M0-DCs were co-cultured with CFSE-labeled naive CD4+ T cells to evaluate T cell proliferation and differentiation. The effects of HMGB1 on DCs activation, cytokine secretion, and autophagy-related protein expression were assessed after treatment with autophagy regulators.

RESULTS

The model group showed significantly elevated levels of HMGB1 in serum and lung tissues, accompanied by upregulated activation markers of DCs and increased expression of autophagy-related protein LC3. HMGB1 stimulation significantly enhanced the activation of M0-DCs and the secretion of pro-inflammatory cytokines, promoting the proliferation of CD4+ T cells and their differentiation into Th1 and Th17 subsets. Rapamycin, which enhances autophagy, potentiated HMGB1-mediated DC activation, while 3-MA, which inhibits autophagy, suppressed the effects of HMGB1, further influencing CD4+ T cell differentiation.

CONCLUSION

HMGB1 modulates DC autophagy, thereby affecting their activation and immune responses of CD4+ T cells in bleomycin-induced PF. Targeting HMGB1 and the autophagy pathway may provide new strategies for the treatment of PF.

摘要

背景

高迁移率族蛋白B1（HMGB1）在炎症和自噬中的作用日益受到关注；然而，其对树突状细胞（DCs）活化及自噬的影响仍不清楚。本研究旨在探讨HMGB1对博来霉素诱导的肺纤维化（PF）小鼠模型中DC活化、自噬及其对CD4⁺T细胞反应的影响。

方法

将30只小鼠随机分为对照组和模型组。模型组通过气管内注射博来霉素诱导PF。采用流式细胞术检测DC表面标志物，蛋白质免疫印迹法评估自噬相关蛋白LC3的表达。使用磁珠分选肺DCs和外周血CD14⁺单核细胞并分化为M0-DCs，然后进行HMGB1刺激实验以评估活化和细胞因子分泌。将经HMGB1刺激或未处理的M0-DCs与羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）标记的初始CD4⁺T细胞共培养，以评估T细胞增殖和分化。在用自噬调节剂处理后，评估HMGB1对DCs活化、细胞因子分泌和自噬相关蛋白表达的影响。

结果

模型组血清和肺组织中HMGB1水平显著升高，同时DCs活化标志物上调，自噬相关蛋白LC3表达增加。HMGB1刺激显著增强M0-DCs的活化和促炎细胞因子的分泌，促进CD4⁺T细胞增殖并分化为Th1和Th17亚群。增强自噬的雷帕霉素增强了HMGB1介导的DC活化，而抑制自噬的3-甲基腺嘌呤（3-MA）抑制了HMGB1的作用，进一步影响CD4⁺T细胞分化。