2016
DOI: 10.1016/j.coi.2016.03.008
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Deep sequencing and human antibody repertoire analysis

Abstract: In the past decade, high-throughput DNA sequencing (HTS) methods and improved approaches for isolating antigen-specific B cells and their antibody genes have been applied in many areas of human immunology. This work has greatly increased our understanding of human antibody repertoires and the specific clones responsible for protective immunity or immune-mediated pathogenesis. Although the principles underlying selection of individual B cell clones in the intact immune system are still under investigation, the … Show more

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Cited by 42 publications
(41 citation statements)
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“…To assess the spectrum of memory cell specification across the CD45RB and CD27 compartments, we prospectively isolated the four quadrants of the CD27 x CD45RB biaxial from healthy, human CD38lo/-B cells (n=2), extracted genomic DNA (gDNA), and sequenced the IgH loci by NGS for immune repertoire analysis (see Methods, Figure 3F). As a proxy for antigen exposure, we measured the donor-pooled mutation frequency of nucleotides in the IgH loci outside of the complementaritydetermining region 3 (CDR3; Figure 3G; Boyd and Crowe, 2016). As expected, CD27+ cells had a relatively high mutational burden, acquired through somatic hypermutation (SHM) after exposure to antigen.…”
mentioning
confidence: 84%
“…To assess the spectrum of memory cell specification across the CD45RB and CD27 compartments, we prospectively isolated the four quadrants of the CD27 x CD45RB biaxial from healthy, human CD38lo/-B cells (n=2), extracted genomic DNA (gDNA), and sequenced the IgH loci by NGS for immune repertoire analysis (see Methods, Figure 3F). As a proxy for antigen exposure, we measured the donor-pooled mutation frequency of nucleotides in the IgH loci outside of the complementaritydetermining region 3 (CDR3; Figure 3G; Boyd and Crowe, 2016). As expected, CD27+ cells had a relatively high mutational burden, acquired through somatic hypermutation (SHM) after exposure to antigen.…”
mentioning
confidence: 84%
“…There are several strategies for reconstructing TCR or BCR repertoires, and the best choice depends on the question of interest (Fig. ).…”
Section: Hts‐based Methods Of Immune Repertoire Profilingmentioning
confidence: 99%
“…Of the early next‐gen platforms 454 was typically favored70, 71, 72, 73 over early Illumina or SOLiD in antibody analysis because of capacity to produce longer reads that could also allow class/subclass determination. Methods using paired end Illumina sequencing have advanced, however, allowing the capture of longer reads and sequencing of the full variable region and subclass isotyping with certain 2x 300 bp paired end sequencing methods 74.…”
Section: Repertoire Analysis Approachesmentioning
confidence: 99%