Next Generation Sequencing Facilitates Quantitative Analysis of IgG or IgE producing B cells

Emergence of IgE antibodies that bind allergens with high affinity is highly correlated with the severity of anaphylaxis. However, the underlying molecular mechanisms by which high-affinity IgE is generated remain poorly understood. We examined the effects of cytokines in IgE class switch using in-vitro B cell activation system Overall design: B cells from spleen of IgE reporter mice (Verigem) were cultured on the CD154+ fibroblast cell line expressing CD40L and BAFF, NB21.2D9 feeder cells in the presence of recombinant murine IL-4, or IL-21 and/or IL-13. IgG+ or IgE reporter positive B cells were sorted and analyzed the transcriptome using Illumina HiSeq1500.