Structure and sequence variation of the canine perforin gene.
Lymphocyte-mediated cytotoxicity is essential to control viral infections, limit lymphocyte expansion and activation, and survey for malignant cells. Humans with defects in lymphocyte cytotoxicity have reduced perforin function resulting in uncontrolled lymphocyte expansion, leading to excessive histiocyte activation and a hemophagocytic disorder. Dog breeds such as Bernese mountain dogs (BMD) have a high incidence of reactive and malignant diseases affecting histiocytes. This study addressed the hypothesis that changes in the perforin gene contribute to the development of hemophagocytic histiocytic sarcoma (HHS) in BMD. Canine perforin DNA was amplified and sequenced through multiple PCR assays from healthy and diseased dogs, and the gene structure determined by rapid amplification of cDNA ends. The coding component of the gene consists of 1679bp, with two exons of 536bp and 1143bp separated by an intron of 865bp. Gene configuration and location differ from that in other species although the coding sequence is highly conserved. Three silent single nucleotide polymorphisms (SNP) were identified. Analysis of their distribution indicated a consistent genotype among 6 middle-aged to older BMD without histiocytic diseases. Among samples from 10 dogs with HHS and 10 without histiocytic diseases SNP occurred with variable frequency. It was concluded that changes in the amino acid sequence of perforin were not associated with HHS but that a constellation of SNP may characterize BMD without histiocytic disease. Investigation of more dogs is required to confirm a specific genotype. Future studies should focus on the potential contribution of reduced perforin expression and/or function to HHS in dogs.
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Department of Pathobiology, University of Guelph, Guelph, ON N1G 2W1, Canada.
Wen
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Moore
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Bienzle
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D000818:Animals; D001483:Base Sequence; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004283:Dog Diseases / Q000235:genetics / Q000276:immunology; D004285:Dogs / Q000235:genetics* / Q000276:immunology*; D005091:Exons; D054747:Histiocytic Sarcoma / Q000235:genetics / Q000276:immunology / Q000662:veterinary; D006801:Humans; D007438:Introns; D051359:Lymphohistiocytosis, Hemophagocytic / Q000235:genetics / Q000276:immunology / Q000662:veterinary; D051379:Mice; D054353:Perforin / Q000235:genetics*; D010802:Phylogeny; D020641:Polymorphism, Single Nucleotide; D013045:Species Specificity
D017931:DNA Primers; D054353:Perforin