Robust T- and B-cell clonotype detection using simultaneous targeted mRNA and immune repertoire profiling of single cells

Abstract

Abstract The key cell types contributing to the diversity of the immune repertoire are the T cells, with their T-cell receptor (TCR) and B cells with their B-cell receptor (BCR). During T- and B-cell development, the cell undergoes recombination of the germline gene segments, variable (V), diversity (D), and joining (J), also called V(D)J recombination. We developed a single cell clonotype detection assay for the BD Rhapsody™ Single-Cell Analysis System capable of processing thousands of single cells at once. We aimed to capture ~20,000 peripheral blood mononuclear cells from each individual donor (total of five) in a BD Rhapsody™ cartridge and took a sub-set of ~3,000 cells for immune repertoire profiling analysis of the complementarity determining region 3 (CDR3) of the variable regions of TCRα/TCRβ and the IgG heavy/light chains. The BD Rhapsody® Human Immune Response Panel for targeted mRNA gene expression analysis was used to classify the lymphocytes and monocyte populations. Results demonstrate that the BD Rhapsody™ VDJ assay can detect clonotype expression differences among different donors with >80% sensitivity and specificity. The abundance of TCR and BCR transcripts was determined by the addition of a unique molecular index to each mRNA molecule. Overall, the current technology demonstrates the ability to evaluate immune populations using single cell next generation sequencing. This assay could be applied not only to study the immune repertoire but to understand the biology of immune-related diseases and immune response to therapies. For Research Use Only. Not for use in diagnostic or therapeutic procedures. BD, the BD Logo and Rhapsody are trademarks of Becton, Dickinson and Company or its affiliates. © 2019 BD. All rights reserved.