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1.
Vet Comp Oncol ; 13(2): 77-88, 2015 Jun.
Article in English | MEDLINE | ID: mdl-23253146

ABSTRACT

Resistance to Escherichia coli l-asparaginase in canine lymphoma occurs frequently with repeated administration, a phenomenon often attributed, without substantiation, to the induction of neutralizing antibodies. To test the hypothesis that treated dogs develop antibodies against the drug, we created an enzyme-linked immunosorbent assay (ELISA) to measure plasma anti-asparaginase immunoglobulin G responses. Using samples from dogs that had received multiple doses, specific reactivity against l-asparaginase was demonstrated, while naïve patients' samples were negative. The optimized ELISA appeared sensitive, with endpoint titers >1 600 000 in positive control dogs. Intra- and inter-assay coefficients of variation were 3.6 and 14.5%. The assay was supported by the observation that ELISA-positive plasma could immunoprecipitate asparaginase activity. When clinical patients were evaluated, 3/10 dogs developed titers after a single injection; with repeated administration, 4/7 dogs were positive. l-asparaginase antibodies showed reduced binding to the PEGylated drug formulation. The ELISA should prove useful in investigating the potential correlation of antibody responses with resistance.


Subject(s)
Antibodies/blood , Antineoplastic Agents/therapeutic use , Asparaginase/therapeutic use , Dog Diseases/drug therapy , Escherichia coli/enzymology , Lymphoma/veterinary , Animals , Antineoplastic Agents/immunology , Asparaginase/immunology , Dog Diseases/blood , Dog Diseases/immunology , Dogs , Enzyme-Linked Immunosorbent Assay/veterinary , Immunoglobulin G/blood , Lymphoma/blood , Lymphoma/drug therapy , Lymphoma/immunology , Reproducibility of Results , Sensitivity and Specificity
2.
Tissue Antigens ; 80(2): 175-83, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22571293

ABSTRACT

In the dog, previous analyses of major histocompatibility complex class I genes suggest a single polymorphic locus, dog leukocyte antigen (DLA)-88. While 51 alleles have been reported, estimates of prevalence have not been made. We hypothesized that, within a breed, DLA-88 diversity would be restricted, and one or more dominant alleles could be identified. Accordingly, we determined allele usage in 47 Golden Retrievers and 39 Boxers. In each population, 10 alleles were found; 4 were shared. Seven novel alleles were identified. DLA-88*05101 and *50801 predominated in Golden Retrievers, while most Boxers carried *03401. In these breeds, DLA-88 polymorphisms are limited and largely non-overlapping. The finding of highly prevalent alleles fulfills an important prerequisite for studying canine CD8+ T-cell responses.


Subject(s)
CD8-Positive T-Lymphocytes/immunology , Dogs/immunology , Histocompatibility Antigens Class I/genetics , Polymorphism, Genetic , Alleles , Amino Acid Sequence , Animals , Breeding , CD8-Positive T-Lymphocytes/cytology , CD8-Positive T-Lymphocytes/metabolism , Dogs/genetics , Gene Frequency , Genetic Loci , Haplotypes , Histocompatibility Antigens Class I/immunology , Homozygote , Models, Molecular , Molecular Sequence Data , Phylogeny , Sequence Alignment
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