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Related Experiment Video

Updated: Jun 12, 2025

T and B Cell Receptor Immune Repertoire Analysis using Next-generation Sequencing
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Full-Length Immune Repertoire Reconstruction and Profiling at the Transcriptome Level Using Long-Read Sequencing.

Xuanmei Luo1,2, Lili Zhang1, Yifei Li1

  • 1Clinical Biobank, Beijing Hospital, National Center of Gerontology, National Health Commission, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing, China.

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Summary

Full-length immune repertoire sequencing (FLIRseq) offers a superior method for analyzing V(D)J recombination and immunoglobulin classes. This advanced technique aids in understanding disease mechanisms and personalizing cell therapies.

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Area of Science:

  • Immunology
  • Genomics
  • Bioinformatics

Background:

  • Traditional short-read sequencing struggles with the diverse immune repertoire (IR).
  • Reconstructing full-length IR is crucial for understanding immune responses and diseases.

Purpose of the Study:

  • To develop and validate a novel workflow for full-length immune repertoire sequencing (FLIRseq).
  • To assess the accuracy, quantification, and comprehensiveness of FLIRseq compared to existing methods.
  • To analyze immune repertoires in various patient cohorts.

Main Methods:

  • Developed FLIRseq workflow using linear rolling circle amplification and nanopore sequencing.
  • Validated accuracy and quantification against plasmid mixtures and short-read BCR/TCR-seq.
  • Applied FLIRseq to analyze IRs in patients with leukemia, allergic diseases, psoriasis, and prostate cancer.

Main Results:

  • FLIRseq demonstrated higher accuracy (lower mismatch/gap rates, higher identity rates) than standard nanopore sequencing.
  • FLIRseq provided more comprehensive data, including CDR3, isotype, and rare V genes, surpassing BCR/TCR-seq.
  • Observed significant changes in clonotype diversity and abnormal BCR/TCR percentages in leukemia remission.
  • Gained insights into V(D)J recombination and immunoglobulin classes in allergic diseases and psoriasis.
  • Identified consistent AlphaFold2-predicted protein structures for T-cell receptor beta chains in psoriatic tissues.

Conclusions:

  • FLIRseq enables unbiased, comprehensive analysis of V(D)J recombination and immunoglobulin classes.
  • FLIRseq aids in characterizing pathogenic mechanisms and monitoring minimal residual disease.
  • FLIRseq supports the customization of adoptive cell therapies.