GSE114309
Single cell paired heavy- and light-chain immunoglobulin sequencing of sorted Lyme disease plasmablasts
Lyme disease (Borrelia burgdorferi infection) is increasingly recognized as a significant source of morbidity world-wide. Here, we investigated B cell responses to Lyme disease through barcode-enabled single cell sequencing of activated B cells (plasmablasts) sorted from PBMCs. Bulk immunglobulin heavy-chain sequencing from this project has also been separately deposited. Additional information regarding patient characteristics and overlap with other data from the SLICE study is available upon request. Overall design: Single cell sequencing of plasmablast antibody genes was performed as previously described (Blum 2018, DOI: 10.1002/eji.201747460) Lyme disease timepoints: Visit 1, Untreated acute infection; Visit 3, one month after completion of treatment; Visit 5, six months after treatment; Visit 7, two years after treatment. Healthy control timepoints: Visit H1, initial visit; Visit H2, six months after initial; Visit H3, one year after initial visit. Plasmablasts were single-cell sorted into the wells of a 96-well plate (1-16 plates per timepoint, per subject) and cDNA waere tagged with unique well-ID barcodes by template switching during reverse transcription. Wells were pooled from within each plate, pooled samples were amplified with immunoglobulin gene-specific primers, and plate-specific index sequences added during library preparation Libraries of up to 48 plates per run were sequenced with MiSeq 2x330 paired-end sequencing