Immune Repertoire Sequencing Using Molecular Identifiers Enables Accurate Clonality Discovery and Clone Size...

Area of Science:

• Immunology
• Molecular Biology
• Bioinformatics

Background:

• Unique molecular identifiers (MIDs) enhance immune repertoire sequencing (IR-seq) accuracy for antibody sequencing.
• Quantifying T cell receptor (TCR) RNA and estimating clone size in T cell repertoire sequencing (TCR-seq) has been challenging.
• This limitation hinders clinical applications like minimal residual disease detection and vaccine efficacy assessment.

Purpose of the Study:

• To develop a method for quantitatively studying TCR RNA molecule copy number and clonality in T cells.
• To address the limitations in detecting rare T cells and estimating clonal expansion in TCR-seq.
• To enable broader applications of TCR-seq in clinical and research settings.

Main Methods:

• Development of the MID Clustering-based Immune Repertoire Sequencing (MIDCIRS) method.
• Application of MID sub-clustering to eliminate erroneous sequences.
• Quantitative analysis of TCR RNA molecule copy number and T cell clonality.

Main Results:

• MIDCIRS requires MID sub-clustering for accurate sequence elimination.
• MIDCIRS achieves sensitive detection of rare T cells (1 in 1 million naive T cells).
• MIDCIRS accurately estimates the degree of T cell clonal expansion.

Conclusions:

• MIDCIRS provides accurate, sensitive, and wide dynamic range TCR-seq.
• The method establishes a foundation for advanced applications in basic research.
• MIDCIRS supports future clinical applications of TCR repertoire analysis.