Characterization of the canine immunoglobulin heavy chain repertoire by next generation sequencing

Mei Hua Hwang, Nikos Darzentas, Dorothee Bienzle, Peter F Moore, Jodi Morrison, Stefan M. Keller

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The ability to mount adaptive immune responses to a diverse array of pathogens is essential to maintaining the health of an individual. The outcome of adaptive immune responses is influenced by the pool of available lymphocyte antigen receptors. Understanding the composition and dynamics of immune repertoires is hence of relevance to characterizing physiologic immunological processes as well as understanding disease pathogenesis. The dog is increasingly recognized as a model for human disease. The objective of this study was to utilize NGS for comprehensive and unbiased analysis of the IGH repertoire in healthy dogs. First, the IGH locus was searched in silico for previously unidentified genes. Second, IGH transcripts from major lymphoid organs were amplified using a 5′RACE approach without V/J primer bias. Third, amplicons were sequenced on an Illumina MiSeq platform, and data were analyzed using the ARResT/Interrogate platform. Data analysis included V/J usage, V-J pairing biases, isotype frequency, CDR3 diversity, convergent recombination, and public repertoires. The results of this study provide a comprehensive IGH repertoire analysis for healthy dogs. These data will allow further improvement of V/J gene-specific primer sets and will serve as baseline for future studies investigating immune repertoires in health and disease.

Original languageEnglish (US)
Pages (from-to)181-190
Number of pages10
JournalVeterinary Immunology and Immunopathology
Volume202
DOIs
StatePublished - Aug 1 2018

Fingerprint

immunoglobulin heavy chains
Immunoglobulin Heavy Chains
Canidae
Adaptive Immunity
Dogs
dogs
lymphocyte antigens
Antigen Receptors
Health
human diseases
Computer Simulation
Genetic Recombination
Genes
data analysis
genes
pathogenesis
Lymphocytes
receptors
loci
pathogens

Keywords

  • Antigen receptor
  • CDR3
  • Dog
  • IGH
  • Rearrangement
  • Repertoire analysis

ASJC Scopus subject areas

  • Immunology
  • veterinary(all)

Cite this

Characterization of the canine immunoglobulin heavy chain repertoire by next generation sequencing. / Hwang, Mei Hua; Darzentas, Nikos; Bienzle, Dorothee; Moore, Peter F; Morrison, Jodi; Keller, Stefan M.

In: Veterinary Immunology and Immunopathology, Vol. 202, 01.08.2018, p. 181-190.

Research output: Contribution to journalArticle

Hwang, Mei Hua ; Darzentas, Nikos ; Bienzle, Dorothee ; Moore, Peter F ; Morrison, Jodi ; Keller, Stefan M. / Characterization of the canine immunoglobulin heavy chain repertoire by next generation sequencing. In: Veterinary Immunology and Immunopathology. 2018 ; Vol. 202. pp. 181-190.
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