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Single-Cell-Based High-Throughput Ig and TCR Repertoire Sequencing Analysis in Rhesus Macaques.

Evan S Walsh1,2, Tammy S Tollison1, Hayden N Brochu1,2

  • 1Department of Molecular Biomedical Sciences, North Carolina State University College of Veterinary Medicine, Raleigh, NC.

Journal of Immunology (Baltimore, Md. : 1950)
|January 6, 2022
PubMed
Summary
This summary is machine-generated.

Researchers developed new assays for high-throughput single-cell immune repertoire sequencing in rhesus macaques. This enables detailed analysis of immunoglobulin (Ig) and T cell receptor (TCR) repertoires in this key model organism.

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Area of Science:

  • Immunology
  • Genomics
  • Microfluidics

Background:

  • High-throughput sequencing of immune receptors is crucial for understanding adaptive immunity.
  • Rhesus macaques are vital model organisms, but lack advanced single-cell immune repertoire sequencing.
  • Polyclonal immunoglobulin (Ig) and T cell receptor (TCR) complexity hinders macaque studies.

Purpose of the Study:

  • To develop and validate high-throughput single-cell immune repertoire sequencing assays for rhesus macaques.
  • To enable comprehensive analysis of Ig and TCR repertoires in non-human primates.
  • To integrate immune repertoire data with single-cell gene expression.

Main Methods:

  • Designed custom primers for all known rhesus macaque Ig and TCR isotypes and chains.
  • Utilized a commercial solution for single-cell immune repertoire profiling.
  • Sequenced repertoires from over 60,000 cells (PBMCs, splenocytes, sorted B and T cells).

Main Results:

  • Successfully recovered all Ig isotypes and TCR chains from single cells.
  • Enabled measurement of clonal expansion in T cells.
  • Paired Ig and TCR repertoires with gene expression profiles from the same cells.

Conclusions:

  • Established the first high-throughput single-cell immune repertoire sequencing assays for rhesus macaques.
  • This breakthrough facilitates deeper investigation of adaptive immunity in a critical primate model.
  • Opens new avenues for studying immune responses and diseases in rhesus macaques.