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Vet Immunol Immunopathol. 2018 Aug;202:153-163. doi: 10.1016/j.vetimm.2018.07.001. Epub 2018 Jul 7.

Identification of genetic variation in equine collagenous lectins using targeted resequencing.

Author information

1
Department of Pathobiology, University of Guelph, Guelph, Ontario, N1E 2W1, Canada. Electronic address: rfrase03@uoguelph.ca.
2
Department of Clinical Studies, University of Guelph, Guelph, Ontario, N1E 2W1, Canada. Electronic address: larroyo@uoguelph.ca.
3
Department of Pathobiology, University of Guelph, Guelph, Ontario, N1E 2W1, Canada. Electronic address: ann.meyer@oicr.ca.
4
Department of Pathobiology, University of Guelph, Guelph, Ontario, N1E 2W1, Canada. Electronic address: blillie@uoguelph.ca.

Abstract

Collagenous lectins are a family of soluble pattern recognition receptors that play an important role in innate immune resistance to infectious disease. Through recognition of carbohydrate motifs on the surface of pathogens, some collagenous lectins can activate the lectin pathway of complement, providing an effective means of host defense. Genetic polymorphisms in collagenous lectins have been shown in several species to predispose animals to a variety of infectious diseases. Infectious diseases are an important cause of morbidity in horses, however little is known regarding the role of equine collagenous lectins. Using a high-throughput, targeted re-sequencing approach, the relationship between genetic variation in equine collagenous lectin genes and susceptibility to disease was investigated. DNA was isolated from tissues obtained from horses submitted for post-mortem examination. Animals were divided into two populations, those with infectious or autoinflammatory diseases (n = 37) and those without (n = 52), and then subdivided by dominant pathological process for a total of 21 pools, each containing 4-5 horses. DNA was extracted from each horse and pooled in equimolar amounts, and the exons, introns, upstream (approximately 50 kb) and downstream (approximately 3 kb) regulatory regions for the 11 equine collagenous lectin genes and related MASP genes were targeted for re-sequencing. A custom target capture kit was used to prepare a sequencing library, which was sequenced on an Illumina MiSeq. After implementing quality control filters, 4559 variants were identified. Of these, 92 were present in the coding regions (43 missense, 1 nonsense, and 48 synonymous), 1414 in introns, 3029 in the upstream region, and 240 in the downstream region. In silico analysis of the missense short nucleotide variants identified 12 mutations with potential to disrupt collagenous lectin protein structure or function, 280 mutations located within predicted transcription factor binding sites, and 95 mutations located within predicted microRNA binding elements. Analysis of allelic association identified 113 mutations that segregated between the infectious/autoinflammatory and non-infectious populations. The variants discovered in this experiment represent potential genetic contributors to disease susceptibility of horses, and will serve as candidates for further population-level genotyping. This study contributes to the growing body of evidence that pooled, high-throughput sequencing is a viable strategy for cost-effective variant discovery.

KEYWORDS:

Collagenous lectins; Equine; Infectious diseases; Innate immunity; Next-generation sequencing; Pooled sequencing

PMID:
30078590
DOI:
10.1016/j.vetimm.2018.07.001
[Indexed for MEDLINE]
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