Single-cell sequencing of rhesus Ig and TCR repertoire

Recent advancements in microfluidics and high-throughput sequencing technologies have enabled recovery of paired heavy- and light- chain of immunoglobulins (Ig) and VDJ- and VJ- chains of T cell receptors (TCR), from thousands of single cells simultaneously in humans and mice. Rhesus macaque is one of the most well-studied model organisms for human adaptive immune response. However, due to the complexity of these polyclonal receptors, high-throughput single-cell rhesus immune repertoire sequencing assays are not yet available. Here we designed custom primers that capture all known rhesus macaque Ig and TCR isotypes and chains. These primers are fully compatible with a commercial solution for single cell immune repertoire profiling. Using these rhesus specific assays, we sequenced Ig and TCR repertoires in over 110,000 cells from cryopreserved rhesus PBMC, splenocytes, and FACS-sorted B and T cells. We were able to recover every Ig isotype and TCR chain, measure clonal expansion in proliferating T cells, and pair Ig/TCR repertoires with gene expression profiles of the same single cells. Our results establish the ability to perform high-throughput immune repertoire analysis in rhesus macaques at the single cell level.