Feng Xiu Li, Zheng Yang, Hao Shan Shan, Zhou Guang Fang, Chen Pu Yan
Key Laboratory of Animal Microbiology of China's Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
Protein Pept Lett. 2019;26(5):348-356. doi: 10.2174/0929866526666190228141650.
The Bursa of Fabricius is an acknowledged central humoral immune organ unique to birds, which provides an ideal research model on the immature B cell development.
In this article, our motivation is to study the role on sIgM and establish the molecular basis and functional processes of Bursal Hexapeptide (BHP) in avian immature B cells DT40 cell lines.
In this article, we detected the expressions of sIgM mRNA with qPCR in DT40 cells with BHP treatment, and investigated the gene expression profiles of BHP-treated DT40 cells, employing microarray analyses. Also, to validate the differentially expressed genes, we performed KEGG pathway and Gene Ontology analysis in the BHP-treated DT40 cells. Finally, we comparatively analyzed the similar regulated genes and their involved immune functional processes between DT40 cell and mouse immature B cell line WEHI231 cell with BHP treatment.
Following the proposed framework, we proved that the BHP enhanced the mRNA expression levels of IgM in DT40 cells, and induced 460 upregulated genes and 460 downregulated genes in BHP-treated DT40 cells. The pathway analysis showed that the differentially regulated genes in DT40 cell line with BHP treatment were involved in 12 enrichment pathways, in which Toll-like receptor signaling pathway was the vital pathways, and cytokine-cytokine receptor interaction and Jak-STAT signaling pathway were another two important pathways in BHP-treated DT40 cells. Moreover, BHP induced the immune related biological processes in BHP-treated DT40 cells, including T cell related, cytokine related, lymphocyte related, and innate immune response GO terms. Finally, the comparatively analysis showed that there were two downregulated genes GATA3 and IFNG to be found co-existed among the differentially expressed genes in BHP-treated DT40 cell and WEHI231 cells, which shared some same immune related functional processes in both cell lines.
After the applying the framework, we proved the inducing roles and the gene expression profiles of BHP on avian immature B cells, and verified some molecular basis from the KEGG and GO analysis. These results provided the insight for mechanism on immature B cell differentiation, and offer the essential direction for the vaccine improvement.
法氏囊是鸟类特有的公认的中枢体液免疫器官,为未成熟B细胞发育提供了理想的研究模型。
本文旨在研究sIgM的作用,并建立法氏囊六肽(BHP)在禽类未成熟B细胞DT40细胞系中的分子基础和功能过程。
本文通过qPCR检测BHP处理的DT40细胞中sIgM mRNA的表达,并采用微阵列分析研究BHP处理的DT40细胞的基因表达谱。此外,为了验证差异表达基因,我们对BHP处理的DT40细胞进行了KEGG通路和基因本体分析。最后,我们比较分析了BHP处理的DT40细胞和小鼠未成熟B细胞系WEHI231细胞中相似的调控基因及其参与的免疫功能过程。
按照所提出的框架,我们证明了BHP增强了DT40细胞中IgM的mRNA表达水平,并在BHP处理的DT40细胞中诱导了460个上调基因和460个下调基因。通路分析表明,BHP处理的DT40细胞系中差异调控的基因涉及12条富集通路,其中Toll样受体信号通路是重要通路,细胞因子-细胞因子受体相互作用和Jak-STAT信号通路是BHP处理的DT40细胞中的另外两条重要通路。此外,BHP在BHP处理的DT40细胞中诱导了免疫相关的生物学过程,包括与T细胞相关、细胞因子相关、淋巴细胞相关和先天免疫应答的基因本体术语。最后,比较分析表明,在BHP处理的DT40细胞和WEHI231细胞的差异表达基因中,有两个下调基因GATA3和IFNG共存,它们在两个细胞系中共享一些相同的免疫相关功能过程。
应用该框架后,我们证明了BHP对禽类未成熟B细胞的诱导作用和基因表达谱,并通过KEGG和基因本体分析验证了一些分子基础。这些结果为未成熟B细胞分化机制提供了见解,并为疫苗改进提供了重要方向。