Ruiz-Lafuente Natalia, Muro Manuel, Minguela Alfredo, Parrado Antonio
Servicio de Inmunología, Hospital Clínico Universitario Virgen de la Arrixaca, Instituto Murciano de Investigación Biosanitaria (IMIB-Arrixaca), El Palmar, 30120 Murcia, Spain.
Biochem Biophys Rep. 2018 Oct 6;16:56-61. doi: 10.1016/j.bbrep.2018.09.007. eCollection 2018 Dec.
The Th2 cytokine IL-4 triggers a signaling cascade which activates transcription by STAT6. The goals of the present study are to define the transcriptomic response of mouse spleen B cells (mSBC) to IL-4 used as single stimulus, its specificity compared to human peripheral blood B cells (hPBBC) and to mouse spleen T cells (mSTC), and the pathways affected. Oligonucleotide-based microarrays were performed using two references, the untreated sample and the cells cultured without IL-4, an experimental design which reduces the potential confounding effect of cellular stress during culture. Specificity was addressed by comparing the response of mSBC and our previously published study on hPBBC, of similar design, and a study by other authors on mSTC. We detected an mSBC-specific response (including novel genes, e.g., Sertad4, Lifr, Pmepa1, Epcam, Tbxas1; and common genes, e.g., Usp2, Cst7, Grtp1, and Casp6), an hPBBC-specific response (e.g., CCL17, MTCL1, GCSAM, HOMER2, IL2RA), and a common mSBC/hPBBC response (e.g., CISH, NFIL3, SOCS1, VDR, CDH1). In contrast, the mSBC and mSTC responses were largely divergent. Gene set enrichment analysis (GSEA) was applied for the first time to identify the pathways affected. Both in mSBC and hPBBC, IL-4 activated Myc, the transcriptional machinery itself, cell cycle, mitochondria and respiratory chain, ribosome, proteasome and antigen presentation, and Wnt signaling, and inhibited GPCR signaling. However, significant differences were found in histone demethylation, Nod signaling, and Rho signaling, which were downregulated in mSBC, and in chromatin condensation, which was downregulated in hPBBC. These findings may have therapeutic implications for the treatment of allergic diseases and parasitic infections.
Th2细胞因子白细胞介素-4(IL-4)引发信号级联反应,通过信号转导和转录激活因子6(STAT6)激活转录。本研究的目的是确定小鼠脾脏B细胞(mSBC)对作为单一刺激物的IL-4的转录组反应,与人类外周血B细胞(hPBBC)和小鼠脾脏T细胞(mSTC)相比其特异性,以及受影响的信号通路。使用两种对照进行基于寡核苷酸的微阵列分析,即未经处理的样本和在无IL-4条件下培养的细胞,这种实验设计减少了培养过程中细胞应激的潜在混杂效应。通过比较mSBC的反应与我们之前发表的关于hPBBC的类似设计研究,以及其他作者关于mSTC的研究,来探讨特异性。我们检测到mSBC特异性反应(包括新基因,如Sertad4、Lifr、Pmepa1、Epcam、Tbxas1;以及常见基因,如Usp2、Cst7、Grtp1和Casp6)、hPBBC特异性反应(如CCL17、MTCL1、GCSAM、HOMER2、IL2RA)以及mSBC/hPBBC共同反应(如CISH、NFIL3、SOCS1、VDR、CDH1)。相比之下,mSBC和mSTC的反应在很大程度上是不同的。首次应用基因集富集分析(GSEA)来确定受影响的信号通路。在mSBC和hPBBC中,IL-4均激活了Myc、转录机制本身、细胞周期、线粒体和呼吸链、核糖体、蛋白酶体和抗原呈递以及Wnt信号通路,并抑制了GPCR信号通路。然而,在组蛋白去甲基化、Nod信号通路和Rho信号通路方面发现了显著差异,这些在mSBC中下调,而在染色质凝聚方面,hPBBC中下调。这些发现可能对过敏性疾病和寄生虫感染的治疗具有治疗意义。