Abstract 1347: Robust T- and B-cell clonotype detection using simultaneous targeted mRNA and immune repertoire profiling of single cells

Abstract

Abstract The heterogeneity and adaptability of the immune repertoire directs the efficacy of immune response to chronic diseases such as autoimmunity disorders and cancer. This diversity is represented by a considerable number of different receptor sequences. The key cell types contributing to this 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. Each cell has a unique sequence required to produce the antigen specificity, referred to as a clonotype. Here, 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. To test the robustness of the assay, five PBMC samples were tested from different individual donors. We aimed to capture ~20,000 peripheral blood mononuclear cells (PBMC) 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 successfully used to classify the lymphocytes and monocyte populations in the samples. Results demonstrate that the BD Rhapsody™ VDJ assay can detect clonotype expression differences among different donors with >80 % sensitivity and specificity. The addition of a unique molecular index (UMI) to each mRNA molecule, allowed the abundance of TCR and BCR transcripts to be determined via counts of UMIs per clonotype per cell. In conjunction with the BD® Human Single-Cell Multiplexing Kit, we are also able to combine and load samples from multiple donors in a BD Rhapsody™ cartridge without altering the immune repertoire. 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. Citation Format: Ricelle Acob, Tina Truong, Jay Levine, Kai Liu, Devon Jensen, Dennis Prosen, Katherine Lazaruk. Robust T- and B-cell clonotype detection using simultaneous targeted mRNA and immune repertoire profiling of single cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1347.