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T and B Cell Receptor Immune Repertoire Analysis using Next-generation Sequencing
Published on: January 12, 2021
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T and B Cell Receptor Immune Repertoire Analysis using Next-generation Sequencing
Lael Werner1, Chen Dor2, Naomi Salamon3
1Pediatric Gastroenterology Unit, Edmond and Lily Safra Children's Hospital, Sheba Medical Center; Sackler Faculty of Medicine, Tel Aviv University; werner.lael@sheba.health.gov.il.
Journal of Visualized Experiments : Jove
|February 1, 2021
Summary
Next-generation sequencing (NGS) provides a detailed view of T and B cell receptors, revealing immune repertoire changes in various diseases. This protocol facilitates the analysis of adaptive immunity for potential clinical applications.
Area of Science:
- Immunology
- Genomics
Background:
- Adaptive immunity relies on T and B lymphocytes with diverse antigen receptors.
- Immune repertoire heterogeneity arises from random genetic recombination.
- Next-generation sequencing (NGS) offers deep insights into T and B cell receptor repertoires.
Purpose of the Study:
- To provide a detailed protocol for NGS of T and B cell immune repertoires from blood and tissue.
- To enable exploration of specific T and B cells at the nucleotide or amino-acid level.
- To identify dynamic changes in lymphocyte populations and diversity parameters in disease.
Main Methods:
- DNA isolation from blood and tissue samples.
- Library preparation and sequencing using NGS technology.
- Bioinformatic pipeline for basic immune repertoire analysis.
Main Results:
- Demonstrated marked changes in immune repertoire clonality, gene usage, and biophysical properties in various conditions.
- Provided a comprehensive pipeline for immune repertoire sequencing and analysis.
- Enabled exploration of adaptive immune responses at a high resolution.
Conclusions:
- NGS of immune repertoires offers valuable insights into adaptive immunity in health and disease.
- The described method can identify dynamic changes in lymphocyte populations.
- This technique has potential applications in biomarker discovery, risk stratification, and precision medicine.
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