Next Generation Sequencing Facilitates Quantitative Analysis of B cell Transcriptomes

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. To examine function of IL-13 derived from T cells in IgE producing B cell development, we transferred IL-13 sufficient or deficient OT-2 TH2 into IgE reporter (Verigem) mice followed by immunization with NP-OVA. At 7 day post immunization, NP specific IgE producing B cells were sorted and served to single cell transcriptome analysis. NP specific IgE+ B cells of mice were sorted and analyzed the transcriptome using Illumina NovaSeq 6000.

能够高亲和力结合过敏原的免疫球蛋白E（IgE）抗体的产生，与过敏性休克的严重程度高度相关。然而，高亲和力IgE的生成背后的潜在分子机制仍未明确。为探究T细胞来源的白细胞介素13（IL-13）在分泌IgE的B细胞发育过程中的功能，我们将携带IL-13或IL-13缺失的OT-2 2型辅助性T细胞（TH2）转移至IgE报告基因（Verigem）小鼠体内，随后以硝基苯化卵清蛋白（NP-OVA）进行免疫。免疫后第7天，我们分选获得NP特异性IgE阳性B细胞并开展单细胞转录组分析。我们进一步分选了小鼠体内的NP特异性IgE+ B细胞，使用Illumina NovaSeq 6000测序平台对其转录组进行分析。