Lung-resident memory B cells maintain allergic IgE responses in the respiratory tract.

We used single cell RNA sequencing to examine the transcriptome and B cell receptor (BCR) sequences of B cells infiltrating mouse lungs 3 weeks after allergen inhalation to investigate the origins of IgE-producing plasma cells. Overall design: Total non-naïve B cells (CD19+IgD-) were isolated from lungs or lung-draining mediastinal lymph nodes (medLNs) of 3 mice by FACS excluding circulating cells identified by intravascular labeling. Isolated cells from each mouse were hashed with BioLegend TotalSeq-C0301, C0302, and C0303 antibodies prior to pooling. Pooled cells from the lungs or medLNs were partitioned on a 10X Chromium Controller and three libraries were prepared for both samples: gene expression, BCR VDJ, and feature barcode libraries for hashing and demultiplexing biological replicates.