WT mice and livin KO mice were developed for model of AAD. Th2 cell levels in the lung tissues and spleen were assessed by flow cytometry. Also, it was assessed in the culture after exposing to livin inhibitor (Lp-15); the protein and mRNA levels of livin, GATA3 and IL-4 in peripheral blood CD4+ T cells isolated from patients with or without AAD were measured by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting, respectively. Finally, Co-immunoprecipitation (Co-IP) was employed to identify the interaction between livin and GATA3.
Compared with WT mouse, Th2 cell frequency in lung tissues and spleen was significantly decreased in livin KO mouse; after adding Lp-15, the differentiation from Naive CD4+T cells in spleen to Th2 cells was blocked; the protein and mRNA levels of livin, GATA3 and IL-4 in AAD group were higher than that in control group. The levels of livin were positively correlated with IL-4, and GATA3 was also positively correlated with IL-4 and livin. GATA3 was detected in the protein complex co-precipitated with livin antibody, and livin was also detected in the protein complex co-precipitated by GATA3 antibody.
Livin increases the expression of IL-4 and facilitates naive CD4+ T cells to differentiate into Th2 cells, which triggers airway allergy.
将WT小鼠和livinKO小鼠开发为AAD模型。通过流式细胞术评估肺组织和脾脏中的Th2细胞水平。此外,在暴露于Livin抑制剂(Lp-15)后的培养物中对其进行评估;Livin的蛋白质和mRNA水平,通过实时定量聚合酶链反应(RT-qPCR)和蛋白质印迹法测量从患有或不患有AAD的患者中分离的外周血CD4T细胞中的GATA3和IL-4,分别。最后,采用免疫共沉淀(Co-IP)来鉴定livin和GATA3之间的相互作用。
与WT鼠标相比,LivinKO小鼠肺组织和脾脏中的Th2细胞频率明显降低;添加Lp-15后,脾脏中幼稚CD4+T细胞向Th2细胞的分化被阻断;Livin的蛋白和mRNA水平,AAD组GATA3和IL-4高于对照组。Livin与IL-4呈正相关,GATA3与IL-4、Livin呈正相关。在与livin抗体共沉淀的蛋白质复合物中检测到GATA3,在GATA3抗体共沉淀的蛋白质复合物中也检测到livin。
Livin增加IL-4的表达,促进幼稚CD4+T细胞分化为Th2细胞,引发气道过敏.