Tracking B-Cell Repertoires and Clonal Histories in Normal and Malignant Lymphocytes

Nicola J Weston-Bell, Graeme Cowan, Surinder S Sahota

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

Abstract / Description of output

Methods for tracking B-cell repertoires and clonal history in normal and malignant B-cells based on immunoglobulin variable region (IGV) gene analysis have developed rapidly with the advent of massive parallel next-generation sequencing (mpNGS) protocols. mpNGS permits a depth of analysis of IGV genes not hitherto feasible, and presents challenges of bioinformatics analysis, which can be readily met by current pipelines. This strategy offers a potential resolution of B-cell usage at a depth that may capture fully the natural state, in a given biological setting. Conventional methods based on RT-PCR amplification and Sanger sequencing are also available where mpNGS is not accessible. Each method offers distinct advantages. Conventional methods for IGV gene sequencing are readily adaptable to most laboratories and provide an ease of analysis to capture salient features of B-cell use. This chapter describes two methods in detail for analysis of IGV genes, mpNGS and conventional RT-PCR with Sanger sequencing.

Original languageEnglish
Title of host publicationGerminal Centers
Subtitle of host publicationMethods and Protocols
EditorsDinis Pedro Calado
Place of PublicationNew York, NY
PublisherHumana Press
Chapter21
Pages281-301
Number of pages21
Volume1623
ISBN (Electronic)9781493970957
ISBN (Print)9781493970940
DOIs
Publication statusPublished - 7 Jun 2017

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
ISSN (Print)1064-3745

Keywords / Materials (for Non-textual outputs)

  • B-cell malignancy
  • B-cell repertoire
  • Germinal center
  • IGHV gene
  • Massive parallel next-generation sequencing
  • Multiple myeloma
  • Tumor origins

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