Allelic diversity at the <i>DLA‐88</i> locus in Golden Retriever and Boxer breeds is limited

Ross, Peter; Buntzman, Adam; Vincent, Benjamin G.; Grover, Elise N.; Gojanovich, Gregory; Collins, Edward J.; Frelinger, Jeffrey A.; Hess, Paul R.

doi:10.1111/j.1399-0039.2012.01889.x

Cited by 28 publications

(54 citation statements)

References 56 publications

(80 reference statements)

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“…These features of the GSHPMD model favor its use in the rigorous study of cytotoxic immune responses to recombinant dystrophin expressed following regional or systemic gene therapy. Interestingly, intra-breed allelic diversity, while limited, has been identified within DLA-88, a dog leukocyte antigen (DLA) gene that displays the highest polymorphism of the MHC class I loci 42, 43, 44. Therefore, immunological studies within a single dog breed, such as the GSHPMD model, may result in varied responses based on DLA class I haplotype, a result that can be expected in clinical trials due to the highly polymorphic HLAs encoding MHC I 45 .…”

Section: Discussionmentioning

confidence: 99%

Mechanism of Deletion Removing All Dystrophin Exons in a Canine Model for DMD Implicates Concerted Evolution of X Chromosome Pseudogenes

VanBelzen

Malik

Henthorn

et al. 2017

Molecular Therapy - Methods & Clinical Development

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Duchenne muscular dystrophy (DMD) is a lethal, X-linked, muscle-wasting disorder caused by mutations in the large, 2.4-Mb dystrophin gene. The majority of DMD-causing mutations are sporadic, multi-exon, frameshifting deletions, with the potential for variable immunological tolerance to the dystrophin protein from patient to patient. While systemic gene therapy holds promise in the treatment of DMD, immune responses to vectors and transgenes must first be rigorously evaluated in informative preclinical models to ensure patient safety. A widely used canine model for DMD, golden retriever muscular dystrophy, expresses detectable amounts of near full-length dystrophin due to alternative splicing around an intronic point mutation, thereby confounding the interpretation of immune responses to dystrophin-derived gene therapies. Here we characterize a naturally occurring deletion in a dystrophin-null canine, the German shorthaired pointer. The deletion spans 5.6 Mb of the X chromosome and encompasses all coding exons of the DMD and TMEM47 genes. The sequences surrounding the deletion breakpoints are virtually identical, suggesting that the deletion occurred through a homologous recombination event. Interestingly, the deletion breakpoints are within loci that are syntenically conserved among mammals, yet the high homology among this subset of ferritin-like loci is unique to the canine genome, suggesting lineage-specific concerted evolution of these atypical sequence elements.

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Section: Discussionmentioning

confidence: 99%

Mechanism of Deletion Removing All Dystrophin Exons in a Canine Model for DMD Implicates Concerted Evolution of X Chromosome Pseudogenes

VanBelzen

Malik

Henthorn

et al. 2017

Molecular Therapy - Methods & Clinical Development

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“…The range of class I molecules of mice and humans that can be tested has been has been expanded beyond the endogenous alleles of RMA-S and T2 by production of transfectants; accordingly, we sought to determine whether peptide binding at the canine classical class I locus, Dog Leukocyte Antigen (DLA)-88 (Graumann et al, 1998), could be evaluated using this same strategy. An RMA-S clone expressing a prevalent allele, DLA-88*50801 (Ross et al, 2012), was therefore generated. Like the parent line, these cells could discriminate motif-matched and -mismatched peptides in a standard stabilization assay.…”

Section: Introductionmentioning

confidence: 99%

A cell-based MHC stabilization assay for the detection of peptide binding to the canine classical class I molecule, DLA-88

Ross

Holmes

Gojanovich

et al. 2012

Veterinary Immunology and Immunopathology

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Identifying immunodominant CTL epitopes is essential for studying CD8+ T-cell responses in populations, but remains difficult, as peptides within antigens typically are too numerous for all to be synthesized and screened. Instead, to facilitate discovery, in silico scanning of proteins for sequences that match the motif, or binding preferences, of the restricting MHC class I allele – the largest determinant of immunodominance – can be used to predict likely candidates. The high false positive rate with this analysis ideally requires binding confirmation, which is obtained routinely by an assay using cell lines such as RMA-S that have defective transporter associated with antigen processing (TAP) machinery, and consequently, few surface class I molecules. The stabilization and resultant increased life-span of peptide-MHC complexes on the cell surface by the addition of true binders validates their identity. To determine whether a similar assay could be developed for dogs, we transfected a prevalent class I allele, DLA-88*50801, into RMA-S. In the BARC3 clone, the recombinant heavy chain was associated with murine β2-microglobulin, and importantly, could differentiate motif-matched and -mismatched peptides by surface MHC stabilization. This work demonstrates the potential to use RMA-S cells transfected with canine alleles as a tool for CTL epitope discovery in this species.

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“…At this time, it is known that among classical dog MHC class I proteins, DLA-88 is polymorphic and represents 59 alleles [13–18]. We investigated the binding motif of DLA-88*50101, which is common to the popular Boxer and Golden Retriever breeds [18].…”

Section: Discussionmentioning

confidence: 99%

“…We investigated the binding motif of DLA-88*50101, which is common to the popular Boxer and Golden Retriever breeds [18]. Using two human cells lines (C1R and K562) transfected with DLA-88*50101, we were able to isolate 3720 peptides from DLA-88*50101.…”

Section: Discussionmentioning

confidence: 99%

“…In contrast, DLA-88 is a highly polymorphic MHC class Ia gene which most likely encodes a classical MHC molecule [13, 14]. There are 59 DLA-88 alleles known to date [13–18]. All DLA-88 alleles show high polymorphism in exons 2 and 3, which consist of constant and hypervariable regions and code for the peptide-binding groove in the α1 and α2 domains [13, 19].…”

Section: Introductionmentioning

confidence: 99%

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Characterization of the Canine MHC Class I DLA-88*50101 Peptide Binding Motif as a Prerequisite for Canine T Cell Immunotherapy

et al. 2016

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There are limitations in pre-clinical settings using mice as a basis for clinical development in humans. In cancer, similarities exist between humans and dogs; thus, the dog patient can be a link in the transition from laboratory research on mouse models to clinical trials in humans. Knowledge of the peptides presented on MHC molecules is fundamental for the development of highly specific T cell-based immunotherapies. This information is available for human MHC molecules but is absent for the canine MHC. In the present study, we characterized the binding motif of dog leukocyte antigen (DLA) class I allele DLA-88*50101, using human C1R and K562 transfected cells expressing the DLA-88*50101 heavy chain. MHC class I immunoaffinity-purification revealed 3720 DLA-88*50101 derived peptides, which enabled the determination of major anchor positions. The characterized binding motif of DLA-88*50101 was similar to HLA-A*02:01. Peptide binding analyses on HLA-A*02:01 and DLA-88*50101 via flow cytometry showed weak binding of DLA-88*50101 derived peptides to HLA-A*02:01, and vice versa. Our results present for the first time a detailed peptide binding motif of the canine MHC class I allelic product DLA-88*50101. These data support the goal of establishing dogs as a suitable animal model for the evaluation and development of T cell-based cancer immunotherapies, benefiting both dog and human patients.

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Allelic diversity at the DLA‐88 locus in Golden Retriever and Boxer breeds is limited

Cited by 28 publications

References 56 publications

Mechanism of Deletion Removing All Dystrophin Exons in a Canine Model for DMD Implicates Concerted Evolution of X Chromosome Pseudogenes

Mechanism of Deletion Removing All Dystrophin Exons in a Canine Model for DMD Implicates Concerted Evolution of X Chromosome Pseudogenes

A cell-based MHC stabilization assay for the detection of peptide binding to the canine classical class I molecule, DLA-88

Characterization of the Canine MHC Class I DLA-88*50101 Peptide Binding Motif as a Prerequisite for Canine T Cell Immunotherapy

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