Mokada-Gopal Lavanya, Boeser Alexander, Lehmann Christian H K, Drepper Friedel, Dudziak Diana, Warscheid Bettina, Voehringer David
Department of Infection Biology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nuremberg, 91054 Erlangen, Germany.
Faculty of Biology, Department of Biochemistry and Functional Proteomics, University of Freiburg, 79104 Freiburg, Germany.
J Immunol. 2017 May 1;198(9):3737-3745. doi: 10.4049/jimmunol.1601838. Epub 2017 Mar 27.
The transcription factor STAT6 plays a key role in mediating signaling downstream of the receptors for IL-4 and IL-13. In B cells, STAT6 is required for class switch recombination to IgE and for germinal center formation during type 2 immune responses directed against allergens or helminths. In this study, we compared the transcriptomes and proteomes of primary mouse B cells from wild-type and STAT6-deficient mice cultured for 4 d in the presence or absence of IL-4. Microarray analysis revealed that 214 mRNAs were upregulated and 149 were downregulated >3-fold by IL-4 in a STAT6-dependent manner. Across all samples, ∼5000 proteins were identified by label-free quantitative liquid chromatography/mass spectrometry. A total of 149 proteins was found to be differentially expressed >3-fold between IL-4-stimulated wild-type and STAT6 B cells (75 upregulated and 74 downregulated). Comparative analysis of the proteome and transcriptome revealed that expression of these proteins was mainly regulated at the transcriptional level, which argues against a major role for posttranscriptional mechanisms that modulate the STAT6-dependent proteome. Nine proteins were selected for confirmation by flow cytometry or Western blot. We show that CD30, CD79b, SLP-76, DEC205, IL-5Rα, STAT5, and Thy1 are induced by IL-4 in a STAT6-dependent manner. In contrast, Syk and Fc receptor-like 1 were downregulated. This dataset provides a framework for further functional analysis of newly identified IL-4-regulated proteins in B cells that may contribute to germinal center formation and IgE switching in type 2 immunity.
转录因子STAT6在介导白细胞介素-4（IL-4）和白细胞介素-13受体下游的信号传导中起关键作用。在B细胞中，STAT6是2型免疫反应（针对过敏原或蠕虫）期间向IgE的类别转换重组和生发中心形成所必需的。在本研究中，我们比较了在有或无IL-4的情况下培养4天的野生型和STAT6缺陷型小鼠原代B细胞的转录组和蛋白质组。微阵列分析显示，214种mRNA以STAT6依赖的方式被IL-4上调，149种被下调超过3倍。在所有样本中，通过无标记定量液相色谱/质谱法鉴定了约5000种蛋白质。发现在IL-4刺激的野生型和STAT6缺陷型B细胞之间共有149种蛋白质差异表达超过3倍（75种上调，74种下调）。蛋白质组和转录组的比较分析表明，这些蛋白质的表达主要在转录水平受到调控，这与调节STAT6依赖蛋白质组的转录后机制的主要作用相悖。选择了9种蛋白质通过流式细胞术或蛋白质免疫印迹进行验证。我们表明，CD30、CD79b、SLP-76、DEC205、IL-5Rα、STAT5和Thy1以STAT6依赖的方式被IL-4诱导。相反，Syk和Fc受体样蛋白1被下调。该数据集为进一步功能分析新鉴定的B细胞中IL-4调节的蛋白质提供了框架，这些蛋白质可能有助于2型免疫中的生发中心形成和IgE转换。
