Department of Molecular & Experimental Medicine, The Scripps Research Institute, La Jolla, California, United States of America.
PLoS One. 2009 Nov 19;4(11):e7906. doi: 10.1371/journal.pone.0007906.
Alternative splicing of pre-mRNA is a mechanism that increases the protein diversity of a single gene by differential exon inclusion/exclusion during post-transcriptional processing. While alternative splicing is established to occur during lymphocyte activation, little is known about the role it plays during the immune response. Our study is among the first reports of a systematic genome-wide analysis of activated human T and B lymphocytes using whole exon DNA microarrays integrating alternative splicing and differential gene expression. Purified human CD2(+) T or CD19(+) B cells were activated using protocols to model the early events in post-transplant allograft immunity and sampled as a function of time during the process of immune activation. Here we show that 3 distinct classes of alternatively spliced and/or differentially expressed genes change in an ordered manner as a function of immune activation. We mapped our results to function-based canonical pathways and demonstrated that some are populated by only one class of genes, like integrin signaling, while other pathways, such as purine metabolism and T cell receptor signaling, are populated by all three classes of genes. Our studies augment the current view of T and B cell activation in immunity that has been based exclusively upon differential gene expression by providing evidence for a large number of molecular networks populated as a function of time and activation by alternatively spliced genes, many of which are constitutively expressed.
前体 mRNA 的选择性剪接是一种通过转录后加工过程中不同外显子的包含/排除来增加单个基因的蛋白质多样性的机制。虽然选择性剪接在淋巴细胞激活过程中已被确定发生，但对于其在免疫反应中所起的作用知之甚少。我们的研究是首次使用全外显子 DNA 微阵列对激活的人 T 和 B 淋巴细胞进行系统的全基因组分析的报告之一，该分析整合了选择性剪接和差异基因表达。使用模拟移植后同种异体免疫早期事件的方案激活纯化的人 CD2(+) T 或 CD19(+) B 细胞，并在免疫激活过程中随时间采样。在这里，我们表明，3 种不同类型的选择性剪接和/或差异表达基因按免疫激活的顺序发生变化。我们将我们的结果映射到基于功能的经典途径，并证明其中一些途径仅由一类基因组成，如整合素信号，而其他途径，如嘌呤代谢和 T 细胞受体信号，由所有 3 类基因组成。我们的研究通过提供大量作为时间和激活函数的分子网络的证据，增加了目前基于差异基因表达的免疫中 T 和 B 细胞激活的观点，其中许多基因是组成型表达的。
