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. 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