Combined Analysis of Transcriptome and T-Cell Receptor Alpha and Beta (TRA /TRB ) Repertoire in Paucicellular Samples at the Single-Cell Level

Methods Mol Biol. 2022:2453:231-259. doi: 10.1007/978-1-0716-2115-8_14.

Abstract

With the advent of next-generation sequencing (NGS) methodologies, the total repertoires of B and T cells can be disclosed in much more detail than ever before. Even though many of these strategies do provide in-depth and high-resolution information of the immunoglobulin (IG) and/or T-cell receptor (TR) repertoire, one clear disadvantage is that the IG/TR profiles cannot be connected to individual cells. Single-cell technologies do allow to study the IG/TR repertoire at the individual cell level. This is especially relevant in cell samples in which much heterogeneity of the cell population is expected. By combining the IG/TR repertoire with transcriptome data, the reactivity of the B or T cell can be associated with activation or maturation stages. An additional advantage of such single-cell technologies is that the combination of both IG and both TR chains can be studied on a per cell basis, which better reflects the antigen receptor reactivity of cells. Here we present the ICELL8 single-cell method for the parallel analysis of the TR repertoire and transcriptome, which is especially useful in samples that contain relatively few cells.

Keywords: Next-generation sequencing; Repertoire; Single cell; T-cell receptor alpha; T-cell receptor beta; Transcriptome.

MeSH terms

  • High-Throughput Nucleotide Sequencing / methods
  • Immunoglobulins / genetics
  • Receptors, Antigen, T-Cell / genetics
  • Receptors, Antigen, T-Cell, alpha-beta* / genetics
  • Transcriptome*

Substances

  • Immunoglobulins
  • Receptors, Antigen, T-Cell
  • Receptors, Antigen, T-Cell, alpha-beta