GSE200955
Next Generation Sequencing analysis of BCR-induced gene expression in adult and neonatal splenic B cells
Newborns are unable to reach the adult-level humoral immune response partly due to the potent immunoregulatory role of IL-10. IL-10 expression is controlled by various transcription factors including nuclear factor-?B (NF-?B), signal transducer and activator of transcription 3 (STAT3), and GATA binding protein 3 (GATA3) depending on the upstream signaling pathways and cell types. In B cells, IL-10 production has been shown to be activated by B cell receptor (BCR) engagement, CD40 ligand, TLR agonists, IL-1ß and IL-6. We have shown increased IL-10 production by BCR-stimulated neonatal B cells compared to adult counterparts. To gain insight into the underlying mechanisms of enhanced IL-10 production in BCR-stimulated neonatal B cells, we performed RNA sequencing and Gene Set Enrichment Analysis for adult and neonatal B cells following BCR engagement. To identify signaling pathways uniquely activated by neonatal BCR, we compared adult and neonatal B cell data using hallmark gene sets from the Molecular Signatures database (MSigDB) C5 gene ontology (GO) collection. Pathways enriched in neonatal B cells included cytokine receptor signaling pathways leading to Signal Transducer And Activator Of Transcription (STAT) protein activation. We also conducted GSEA using the MSigDB C3 transcription factor targets (TFT) hallmark gene collection to identify transcription factors uniquely activated in neonatal B cells. This analysis revealed that target gene sets for STAT3, STAT5A and STAT5B were highly enriched in neonatal B cells, suggesting that these STAT proteins were activated in neonatal B cells after BCR engagement. Overall design: Total RNA obtained from isolated adult and neonatal splenic CD19+CD43- B cells subjected to 0 or 7 hours ex vivo anti-IgM. 18 samples containing triplicated 6 conditions.