Type 2-polarized memory B cells hold allergen-specific IgE memory
Allergen-specific IgE antibodies mediate allergic pathology in diseases such as allergic rhinitis and food allergy. Memory B cells (MBCs) contribute to circulating IgE by regenerating IgE-producing plasma cells upon allergen encounter. We report a population of type 2 polarized MBCs defined as CD23hi, IL-4Rahi, CD32low at the transcriptional and surface protein levels. Overall design: Memory B cells were negatvely selected from PBMC's from allergics and non-allergics using magnetic coloumns and antibody labelled beads. The MBC enriched fractions were analysed using the 10X Genomics Chromium Instrument and cDNA synthesis kit (10x Genomics) to generate a barcoded cDNA library for single cell RNA-sequencing from approximately 10,000 cells. cDNA library quality was determined using an Agilent Bioanalyzer. Using this library, we performed pair end sequencing on an Illumina NovaSeq6000. 10X Genomics Chromium v2 chemistry