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

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Authors

HWANG M.H. DARZENTAS Nikos BIENZLE D. MOORE P.F. MORRISON J. KELLER S.M.

Year of publication 2018
Type Article in Periodical
Magazine / Source VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
MU Faculty or unit

Central European Institute of Technology

Citation
Doi http://dx.doi.org/10.1016/j.vetimm.2018.07.002
Keywords Dog; IGH; Repertoire analysis; CDR3; Rearrangement; Antigen receptor
Description 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 Illumine 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.
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