c-CBL/LCK/c-JUN/ETS1/CD28 轴通过抑制 Th2 分化来抑制儿童哮喘。

c-CBL/LCK/c-JUN/ETS1/CD28 axis restrains childhood asthma by suppressing Th2 differentiation.

机构信息

Department of Pediatrics, Shengjing Hospital of China Medical University, 36 Sanhao Street, Shenyang, 110004, China.

出版信息

Mol Med. 2024 Sep 28;30(1):164. doi: 10.1186/s10020-024-00872-1.

DOI:10.1186/s10020-024-00872-1

PMID:39342146

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11439220/

Abstract

BACKGROUND

Asthma is a common immune disease with high morbidity in children. Type 2 inflammation is the center of asthma development, and mainly mediated by a subset of CD4 + T cells, T helper 2 (Th2) cells. Excess Th2 differentiation was generally associated with asthmatic attack. Casitas B-lineage lymphoma (c-CBL) was reported to involved in T cell development and databank showed its decreased expression in CD4 + T cells from peripheral blood of asthmatic children. This study aims to investigate the role of c-CBL in childhood asthma and Th2 differentiation, and explore the underlying mechanism.

METHODS

We collected peripheral blood samples from clinical childhood asthma cases and healthy controls, and determined c-CBL expression in CD4 + T cells. Asthma was induced in neonatal mice by ovalbumin (OVA) intraperitoneal injection and aerosol inhalation, and c-CBL expression in CD4 + T cells from peripheral blood and spleen was measured. Gain-of-function experiments was performed to confirm the effects of c-CBL on Th2 differentiation in vitro. Finally, c-CBL was delivered into asthmatic mice via lentivirus infection to verify its effects on experimental asthma.

RESULTS

c-CBL was lowly expressed in CD4 + T cells from asthmatic children than those of healthy controls. Similarly, it was downregulated in CD4 + T cells from peripheral blood and spleen of asthma mice. Overexpression of c-CBL restrained lung pathological injury and type 2 inflammation in experimental asthmatic mice. Gain-of-function experiments demonstrated that c-CBL inhibited Th2 differentiation of CD4 + T cells from healthy children, and mediated the ubiquitination of lymphocyte cell-specific protein-tyrosine kinase (LCK). LCK acted as a kinase to phosphorylate and activate c-JUN, which was predicted to bind promoter sequence of CD28 by bioinformatic analysis. Dual-luciferase reporter assay verified that c-JUN and ETS1 synergically enhanced transcription of CD28, and this transcription activation was aggravated by LCK overexpression.

CONCLUSION

c-CBL alleviated asthma and suppressed Th2 differentiation by facilitating LCK ubiquitination, interrupting c-JUN activation and CD28 expression in vivo and in vitro. c-CBL/LCK/c-JUN/ETS1/CD28 axis was partially involved in childhood asthma, and may provide novel insights for clinical treatment for asthma.

摘要

背景

哮喘是一种常见的儿童免疫性疾病，发病率高。2 型炎症是哮喘发展的中心，主要由一组 CD4+T 细胞，辅助性 T 细胞 2（Th2）细胞介导。Th2 分化过度通常与哮喘发作有关。已报道 Casitas B 细胞淋巴瘤（c-CBL）参与 T 细胞发育，数据库显示其在哮喘儿童外周血 CD4+T 细胞中的表达降低。本研究旨在探讨 c-CBL 在儿童哮喘和 Th2 分化中的作用，并探讨其潜在机制。

方法

我们收集了临床儿童哮喘病例和健康对照者的外周血样本，并测定了 CD4+T 细胞中的 c-CBL 表达。通过卵清蛋白（OVA）腹腔注射和雾化吸入诱导新生小鼠哮喘，并测定外周血和脾 CD4+T 细胞中的 c-CBL 表达。进行功能获得实验以确认 c-CBL 对体外 Th2 分化的影响。最后，通过慢病毒感染将 c-CBL 递送至哮喘小鼠体内，以验证其对实验性哮喘的影响。

结果

哮喘患儿 CD4+T 细胞中的 c-CBL 表达低于健康对照组。同样，哮喘小鼠外周血和脾 CD4+T 细胞中的 c-CBL 表达也下调。过表达 c-CBL 可抑制实验性哮喘小鼠的肺病理损伤和 2 型炎症。功能获得实验表明，c-CBL 抑制了健康儿童 CD4+T 细胞的 Th2 分化，并介导了淋巴细胞特异性蛋白酪氨酸激酶（LCK）的泛素化。LCK 作为一种激酶磷酸化并激活 c-JUN，生物信息学分析预测 c-JUN 与 CD28 启动子序列结合。双荧光素酶报告基因检测证实 c-JUN 和 ETS1 协同增强 CD28 的转录，而过表达 LCK 则加剧了这种转录激活。