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BACKGROUND: Coronary artery disease (CAD), the leading cause of death worldwide, is influenced by both environmental and genetic factors. Although over 250 genetic risk loci have been identified through genome-wide association studies, the specific causal variants and their regulatory mechanisms are still largely unknown, particularly in disease-relevant cell types such as macrophages. METHODS: We utilized single-cell RNA-seq and single-cell multiomics approaches in primary human monocyte-derived macrophages to explore the transcriptional regulatory network involved in a critical pathogenic event of coronary atherosclerosis-the formation of lipid-laden foam cells. The relative genetic contribution to CAD was assessed by partitioning disease heritability across different macrophage subpopulations. Meta-analysis of single-cell RNA-seq data sets from 38 human atherosclerotic samples was conducted to provide high-resolution cross-referencing to macrophage subpopulations in vivo. RESULTS: We identified 18â 782 cis-regulatory elements by jointly profiling the gene expression and chromatin accessibility of >5000 macrophages. Integration with CAD genome-wide association study data prioritized 121 CAD-related genetic variants and 56 candidate causal genes. We showed that CAD heritability was not uniformly distributed and was particularly enriched in the gene programs of a novel CD52-hi lipid-handling macrophage subpopulation. These CD52-hi macrophages displayed significantly less lipoprotein accumulation and were also found in human atherosclerotic plaques. We investigated the cis-regulatory effect of a risk variant rs10488763 on FDX1, implicating the recruitment of AP-1 and C/EBP-ß in the causal mechanisms at this locus. CONCLUSIONS: Our results provide genetic evidence of the divergent roles of macrophage subsets in atherogenesis and highlight lipid-handling macrophages as a key subpopulation through which genetic variants operate to influence disease. These findings provide an unbiased framework for functional fine-mapping of genome-wide association study results using single-cell multiomics and offer new insights into the genotype-environment interactions underlying atherosclerotic disease.
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Doença da Artéria Coronariana , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Macrófagos , Humanos , Doença da Artéria Coronariana/genética , Doença da Artéria Coronariana/metabolismo , Doença da Artéria Coronariana/patologia , Macrófagos/metabolismo , Fatores de Risco , Análise de Célula Única , Redes Reguladoras de Genes , Masculino , Polimorfismo de Nucleotídeo Único , FemininoRESUMO
A causative role for single nucleotide polymorphisms (SNPs) in many genetic disorders has become evident through numerous genome-wide association studies. However, identification of these common causal variants and the molecular mechanisms underlying these associations remains a major challenge. Differential transcription factor binding at a SNP resulting in altered gene expression is one possible mechanism. Here we apply PWAS ("proteome-wide analysis of SNPs"), a methodology based on quantitative mass spectrometry that enables rapid screening of SNPs for differential transcription factor binding, to 12 SNPs that are highly associated with type 1 diabetes at the IL2RA locus, encoding the interleukin-2 receptor CD25. We report differential, allele-specific binding of the transcription factors RUNX1, LEF1, CREB, and TFAP4 to IL2RA SNPs rs12722508*A, rs12722522*C, rs41295061*A, and rs2104286*A and demonstrate the functional influence of RUNX1 at rs12722508 by reporter gene assay. Thus, PWAS may be able to contribute to our understanding of the molecular consequences of human genetic variability underpinning susceptibility to multi-factorial disease.
Assuntos
Diabetes Mellitus Tipo 1/genética , Subunidade alfa de Receptor de Interleucina-2/genética , Polimorfismo de Nucleotídeo Único/genética , Proteoma , Alelos , Subunidade alfa 2 de Fator de Ligação ao Core/metabolismo , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Regulação da Expressão Gênica , Predisposição Genética para Doença , Genoma Humano , Ensaios de Triagem em Larga Escala , Humanos , Subunidade alfa de Receptor de Interleucina-2/metabolismo , Fator 1 de Ligação ao Facilitador Linfoide/metabolismo , Ligação Proteica , Proteoma/análise , Proteoma/genéticaRESUMO
The chromosome 16p13 region has been associated with several autoimmune diseases, including type 1 diabetes (T1D) and multiple sclerosis (MS). CLEC16A has been reported as the most likely candidate gene in the region, since it contains the most disease-associated single-nucleotide polymorphisms (SNPs), as well as an imunoreceptor tyrosine-based activation motif. However, here we report that intron 19 of CLEC16A, containing the most autoimmune disease-associated SNPs, appears to behave as a regulatory sequence, affecting the expression of a neighbouring gene, DEXI. The CLEC16A alleles that are protective from T1D and MS are associated with increased expression of DEXI, and no other genes in the region, in two independent monocyte gene expression data sets. Critically, using chromosome conformation capture (3C), we identified physical proximity between the DEXI promoter region and intron 19 of CLEC16A, separated by a loop of >150 kb. In reciprocal experiments, a 20 kb fragment of intron 19 of CLEC16A, containing SNPs associated with T1D and MS, as well as with DEXI expression, interacted with the promotor region of DEXI but not with candidate DNA fragments containing other potential causal genes in the region, including CLEC16A. Intron 19 of CLEC16A is highly enriched for transcription-factor-binding events and markers associated with enhancer activity. Taken together, these data indicate that although the causal variants in the 16p13 region lie within CLEC16A, DEXI is an unappreciated autoimmune disease candidate gene, and illustrate the power of the 3C approach in progressing from genome-wide association studies results to candidate causal genes.
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Doenças Autoimunes/genética , Proteínas de Ligação a DNA/genética , DNA/genética , Proteínas de Membrana/genética , Cromossomos Humanos Par 16 , Humanos , Monócitos/metabolismo , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Locos de Características QuantitativasRESUMO
Canine diabetes results from a wide spectrum of clinical pathophysiological processes that cause a similar set of clinical signs. Various causes of insulin deficiency and beta cell loss, insulin resistance, or both characterize the disease, with genetics and environment playing a role. Understanding the genetic and molecular causes of beta cell loss will provide future opportunities for precision medicine, both from a therapeutic and preventative perspective. This review presents current knowledge of the etiology and pathophysiology of canine diabetes, including the importance of disease classification. Examples of potential targets for future precision medicine-based approaches to therapy are discussed.
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Diabetes Mellitus Tipo 2 , Diabetes Mellitus , Doenças do Cão , Resistência à Insulina , Cães , Animais , Diabetes Mellitus/etiologia , Diabetes Mellitus/veterinária , Diabetes Mellitus/diagnóstico , Insulina/uso terapêutico , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/veterinária , Doenças do Cão/genética , Doenças do Cão/terapiaRESUMO
BACKGROUND: Cats presenting with upper urinary tract uroliths (UUTUs) and ureteral obstruction ("obstructive UUTU") are typically younger than cats with idiopathic CKD that often have incidental nephroliths. HYPOTHESIS: Cats with upper urinary tract urolith have 2 clinical phenotypes; a more aggressive phenotype at risk of obstructive UUTU at a young age and a more benign phenotype in older cats, with reduced risk of obstructive UUTU. OBJECTIVES: Identify risk factors for UUTU and for obstructive UUTU. ANIMALS: Eleven thousand four hundred thirty-one cats were referred for care over 10 years; 521 (4.6%) with UUTU. METHODS: Retrospective VetCompass observational cross-sectional study. Multivariable logistic regression models were performed to identify risk factors for a diagnosis of UUTU vs no UUTU and additionally, obstructive UUTU vs nonobstructive UUTU. RESULTS: Risk factors for UUTU included female sex (odds ratio [OR] 1.6, confidence interval [CI] 1.3-1.9; P < .001), British shorthair, Burmese, Persian, Ragdoll or Tonkinese (vs non-purebred ORs 1.92-3.31; P < .001) breed and being ≥4 years (ORs 2.1-3.9; P < .001). Risk factors for obstructive UUTU were female sex (OR 1.8, CI 1.2-2.6; P = .002), having bilateral uroliths (OR 2.0, CI 1.4-2.9; P = .002) and age, with the odds of obstructive UUTU increasing as age at diagnosis of UUTU decreased (≥12 years, reference category; 8-11.9 years, OR 2.7, CI 1.6-4.5; 4-7.9 years, OR 4.1, CI 2.5-7.0; 0-3.9 years, OR 4.3, CI 2.2-8.6; P < 0.001). CONCLUSIONS AND CLINICAL IMPORTANCE: Cats diagnosed with UUTU at a younger age have a more aggressive phenotype with higher risk of obstructive UUTU compared to cats over 12 years of age diagnosed with UUTU.
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Doenças do Gato , Obstrução Ureteral , Cálculos Urinários , Sistema Urinário , Animais , Gatos , Feminino , Masculino , Doenças do Gato/epidemiologia , Estudos Transversais , Estudos Retrospectivos , Fatores de Risco , Reino Unido/epidemiologia , Obstrução Ureteral/epidemiologia , Obstrução Ureteral/veterinária , Cálculos Urinários/veterináriaRESUMO
BACKGROUND: Diabetes mellitus (DM) is an important endocrine disorder in dogs. This study explored prior exposure to glucocorticoids or antibiotic treatment as risk factors for developing DM in dogs attending primary-care VetCompass clinics in the UK. METHODS: A breed frequency matched case-control study nested in a cohort of dogs (n = 480,469) aged 3 years or over was used to explore associations between glucocorticoid and antibiotic exposure and the odds of developing DM. RESULTS: A total of 565 cases and 2179 controls were included. Dogs with DM had over four times the odds of exposure to glucocorticoids within 6 weeks prior to diagnosis (odds ratio [OR] 4.07, 95% confidence interval [CI] 2.41-6.89, p < 0.001) compared to controls within 6 weeks prior to a randomly selected quasi-date of diagnosis. Dogs that had only one unique documented antibiotic course had a decreased odds of developing DM (OR 0.65, 95% CI 0.46-0.91, p = 0.012) compared to dogs that had no documented courses of antibiotics. LIMITATIONS: This study only included selected breeds, so the results may not be generalisable to all dog breeds. CONCLUSIONS: Exposure to glucocorticoids is associated with a substantial increase in the risk of developing DM for the dog breeds included in this analysis.
Assuntos
Diabetes Mellitus , Doenças do Cão , Cães , Animais , Glucocorticoides , Estudos de Casos e Controles , Diabetes Mellitus/veterinária , Fatores de Risco , Reino UnidoRESUMO
Differentiating between canine inflammatory bowel disease (IBD) and intestinal T-cell lymphoma by histopathological examination of endoscopically-derived intestinal biopsies can be challenging and involves an invasive procedure requiring specialized equipment and training. A rapid, non-invasive method of diagnosis, such as blood or faecal analysis for a conserved and stable biomarker, would be a useful adjunct or replacement. Studies on dogs and humans with various types of lymphoma have shown altered microRNA (miRNA) expression patterns in blood, faeces and tissues indicating their potential use as biomarkers of disease. The present study used residual archived endoscopically-derived, formalin-fixed, paraffin-embedded (FFPE) duodenal tissue taken from pet dogs undergoing routine investigation of gastrointestinal disease. The dogs had previously been diagnosed with either normal/minimal intestinal inflammation, severe IBD or intestinal T-cell lymphoma. Next generation sequencing with qPCR validation was used to elucidate differentially expressed miRNAs between groups. Our results show that miRNA can be extracted from archived endoscopically-derived FFPE tissues from the canine duodenum and used to differentiate normal/minimally inflamed canine duodenal tissue from severe lymphoplasmacytic IBD and T-cell lymphoma.
Assuntos
Doenças do Cão , Doenças Inflamatórias Intestinais , Linfoma de Células T , MicroRNAs , Humanos , Cães , Animais , Intestinos/patologia , Doenças Inflamatórias Intestinais/veterinária , Doenças Inflamatórias Intestinais/genética , Doenças Inflamatórias Intestinais/metabolismo , Duodeno/metabolismo , Duodeno/patologia , Linfoma de Células T/veterinária , Biomarcadores/metabolismo , MicroRNAs/metabolismo , Doenças do Cão/patologiaRESUMO
BACKGROUND: The detection of insulin autoantibodies (IAA) aids in the prediction of autoimmune diabetes development. However, the long-standing, gold standard 125I-insulin radiobinding assay (RBA) has low reproducibility between laboratories, long sample processing times and requires the use of newly synthesized radiolabeled insulin for each set of assays. Therefore, a rapid, non-radioactive, and reproducible assay is highly desirable. METHODS: We have developed electrochemiluminescence (ECL)-based assays that fulfill these criteria in the measurement of IAA and anti-insulin antibodies (IA) in non-obese diabetic (NOD) mice and in type 1 diabetic individuals, respectively. Using the murine IAA ECL assay, we examined the correlation between IAA, histopathological insulitis, and blood glucose in a cohort of female NOD mice from 4 up to 36 weeks of age. We developed a human IA ECL assay that we compared to conventional RBA and validated using samples from 34 diabetic and 59 non-diabetic individuals in three independent laboratories. RESULTS: Our ECL assays were rapid and sensitive with a broad dynamic range and low background. In the NOD mouse model, IAA levels measured by ECL were positively correlated with insulitis severity, and the values measured at 8-10 weeks of age were predictive of diabetes onset. Using human serum and plasma samples, our IA ECL assay yielded reproducible and accurate results with an average sensitivity of 84% at 95% specificity with no statistically significant difference between laboratories. CONCLUSIONS: These novel, non-radioactive ECL-based assays should facilitate reliable and fast detection of antibodies to insulin and its precursors sera and plasma in a standardized manner between laboratories in both research and clinical settings. Our next step is to evaluate the human IA assay in the detection of IAA in prediabetic subjects or those at risk of type 1 diabetes and to develop similar assays for other autoantibodies that together are predictive for the diagnosis of this common disorder, in order to improve prediction and facilitate future therapeutic trials.
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Diabetes Mellitus Tipo 1/sangue , Diabetes Mellitus Tipo 1/diagnóstico , Eletroquímica/métodos , Anticorpos Anti-Insulina/sangue , Células Secretoras de Insulina/patologia , Medições Luminescentes/métodos , Animais , Autoanticorpos/sangue , Diabetes Mellitus Tipo 1/patologia , Progressão da Doença , Feminino , Humanos , Células Secretoras de Insulina/metabolismo , Camundongos , Camundongos Endogâmicos NOD , Curva ROC , Ensaio Radioligante , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
This two-part article discusses the mechanisms by which genetic variation can influence the risk of complex diseases, with a focus on canine diabetes mellitus. In Part 1, presented here, the importance of accurate methods for classifying different types of diabetes will be discussed, since this underpins the selection of cases and controls for genetic studies. Part 2 will focus on our current understanding of the genes involved in diabetes risk, and the way in which new genome sequencing technologies are poised to reveal new diabetes genes in veterinary species.
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Diabetes Mellitus/veterinária , Doenças do Cão/genética , Predisposição Genética para Doença , Fenótipo , Animais , Diabetes Mellitus/classificação , Diabetes Mellitus/genética , Doenças do Cão/imunologia , Cães , Variação Genética , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Humanos , Resistência à Insulina , Células Secretoras de Insulina/imunologia , Obesidade/veterinária , Especificidade da EspécieRESUMO
Part 1 of this 2-part review outlined the importance of disease classification in diabetes genetic studies, as well as the ways in which genetic variants may contribute to risk of a complex disease within an individual, or within a particular group of individuals. Part 2, presented here, describes in more detail our current understanding of the genetics of canine diabetes mellitus compared to our knowledge of the human disease. Ongoing work to improve our knowledge, using new technologies, is also introduced.
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Diabetes Mellitus/veterinária , Doenças do Cão/genética , Predisposição Genética para Doença/genética , Animais , Diabetes Mellitus/classificação , Diabetes Mellitus/genética , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/imunologia , Doenças do Cão/classificação , Doenças do Cão/imunologia , Cães , Antígenos HLA/genética , Antígenos HLA/imunologia , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Humanos , Imunidade/genética , Células Secretoras de Insulina/imunologia , Complexo Principal de Histocompatibilidade/genética , MutaçãoRESUMO
OBJECTIVES: The objectives of this study were to validate a commercially available luteinising hormone (LH) cat ELISA, to determine whether the increases in plasma LH concentration that occur after neutering are maintained throughout cats' lives and if other factors such as calendar seasons in both intact and neutered cats, and neutering age in neutered cats, influence plasma LH concentrations. METHODS: Stored plasma samples from client-owned cats were used for the measurement of LH concentrations. Clinical data, including age, sex, age at neutering and medical history, were reviewed. Two populations were included in this study: (1) a senior and geriatric cat population (⩾9 years old), including 18 intact and 18 neutered cats matched for age, sex and month of sample collection; and (2) an adult cat population (2-8 years old), including 45 neutered cats. LH concentrations were measured using a commercially available feline ELISA. RESULTS: Senior and geriatric neutered cats had higher plasma LH concentrations than age-matched intact cats (P <0.001). Calendar season did not influence plasma LH concentrations in the adult (P = 0.727) or senior/geriatric (P = 0.745) cats included in this study. No influence of age at neutering was observed on plasma LH concentrations (P = 0.296). CONCLUSIONS AND RELEVANCE: Neutering causes a significant long-term increase in LH concentrations in cats and further studies are required to determine the consequences on feline health.
Assuntos
Hormônio Luteinizante , Animais , GatosRESUMO
GPR84 is a member of the metabolic G protein-coupled receptor family, and its expression has been described predominantly in immune cells. GPR84 activation is involved in the inflammatory response, but the mechanisms by which it modulates inflammation have been incompletely described. In this study, we investigated GPR84 expression, activation, and function in macrophages to establish the role of the receptor during the inflammatory response. We observed that GPR84 expression in murine tissues is increased by endotoxemia, hyperglycemia, and hypercholesterolemia. Ex vivo studies revealed that GPR84 mRNA expression is increased by LPS and other pro-inflammatory molecules in different murine and human macrophage populations. Likewise, high glucose concentrations and the presence of oxidized LDL increased GPR84 expression in macrophages. Activation of the GPR84 receptor with a selective agonist, 6-(octylamino) pyrimidine-2,4(1H,3H)-dione (6-n-octylaminouracil, 6-OAU), enhanced the expression of phosphorylated Akt, p-ERK, and p65 nuclear translocation under inflammatory conditions and elevated the expression levels of the inflammatory mediators TNFα, IL-6, IL-12B, CCL2, CCL5, and CXCL1. In addition, GPR84 activation triggered increased bacterial adhesion and phagocytosis in macrophages. The enhanced inflammatory response mediated by 6-OAU was not observed in GPR84-/- cells nor in macrophages treated with a selective GPR84 antagonist. Collectively, our results reveal that GPR84 functions as an enhancer of inflammatory signaling in macrophages once inflammation is established. Therefore, molecules that antagonize the GPR84 receptor may be potential therapeutic tools in inflammatory and metabolic diseases.
RESUMO
Despite decades of research in humans and mouse models of disease, substantial gaps remain in our understanding of pathogenic mechanisms underlying the development of type 1 diabetes. Furthermore, translation of therapies from preclinical efforts capable of delaying or halting ß-cell destruction has been limited. Hence, a pressing need exists to identify alternative animal models that reflect human disease. Canine insulin deficiency diabetes is, in some cases, considered to follow autoimmune pathogenesis, similar to NOD mice and humans, characterized by hyperglycemia requiring lifelong exogenous insulin therapy. Also similar to human type 1 diabetes, the canonical canine disorder appears to be increasing in prevalence. Whereas islet architecture in rodents is distinctly different from humans, canine pancreatic endocrine cell distribution is more similar. Differences in breed susceptibility alongside associations with MHC and other canine immune response genes parallel that of different ethnic groups within the human population, a potential benefit over NOD mice. The impact of environment on disease development also favors canine over rodent models. Herein, we consider the potential for canine diabetes to provide valuable insights for human type 1 diabetes in terms of pancreatic histopathology, impairment of ß-cell function and mass, islet inflammation (i.e., insulitis), and autoantibodies specific for ß-cell antigens.
Assuntos
Autoanticorpos/imunologia , Diabetes Mellitus Tipo 1/imunologia , Modelos Animais de Doenças , Cães , Células Secretoras de Insulina/imunologia , Animais , Diabetes Mellitus Tipo 1/tratamento farmacológico , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/patologia , Antígenos HLA/genética , Antígenos de Histocompatibilidade/genética , Humanos , Hipoglicemiantes/uso terapêutico , Inflamação , Insulina/metabolismo , Insulina/uso terapêutico , Secreção de Insulina , Células Secretoras de Insulina/metabolismo , Células Secretoras de Insulina/patologia , Ilhotas Pancreáticas/imunologia , Ilhotas Pancreáticas/patologia , Camundongos , Camundongos Endogâmicos NOD , Pâncreas/patologia , RatosRESUMO
Having gained some clinical experience before pursuing a research career, Lucy Davison is now seeking the best of both worlds as a clinician scientist.
Assuntos
Pesquisa Biomédica , Escolha da Profissão , Medicina Veterinária , HumanosRESUMO
A continuous glucose monitoring system (CGMS) was evaluated in 14 cats with naturally occurring diabetes mellitus. The device measures interstitial fluid glucose continuously, by means of a sensor placed in the subcutaneous tissue. All cats tolerated the device well and a trace was obtained on 15/16 occasions. There was good correlation between the CGMS values and blood glucose concentration measured using a glucometer (r=0.932, P<0.01). Limitations to the use of the CGMS are its working glucose range of 2.2-22.2 mmol/l (40-400 mg/dl) and the need for calibration with a blood glucose measurement at least every 12 h. When compared to a traditional blood glucose curve, the CGMS is minimally invasive, reduces the number of venepunctures necessary to assess the kinetics of insulin therapy in a patient and provides a truly continuous glucose curve.
Assuntos
Automonitorização da Glicemia/veterinária , Glicemia/metabolismo , Doenças do Gato/sangue , Diabetes Mellitus/veterinária , Animais , Automonitorização da Glicemia/métodos , Calibragem , Gatos , Diabetes Mellitus/sangue , Feminino , Masculino , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The presence of anti-insulin antibodies was determined by ELISA in serum samples from 30 diabetic dogs receiving bovine insulin therapy and 30 normoglycaemic dogs. Twenty of the diabetic dogs had significant reactivity to both bovine (heterologous) and porcine (homologous) insulin compared to control dogs. In contrast there was no significant difference between the two populations in reactivity to canine distemper virus (CDV) or canine thyroglobulin. The high degree of correlation between anti-bovine insulin and anti-porcine insulin antibodies suggested cross-reactivity which was confirmed by performing a competition ELISA, with antibody binding to bovine insulin inhibited by pre-incubating serum with porcine insulin. The insulin B-chain, rather than the A-chain was the most reactive component of the insulin molecule although in some cases, diabetics with antibody reactivity to whole insulin protein showed minimal reactivity to the individual subunits. The data suggest that treatment of diabetic dogs with bovine insulin can lead to anti-insulin antibody production. These antibodies cross-react with homologous insulin and recognise conformational as well as linear epitopes.
Assuntos
Diabetes Mellitus/veterinária , Doenças do Cão/imunologia , Anticorpos Anti-Insulina/sangue , Insulina/imunologia , Animais , Bovinos , Reações Cruzadas , Diabetes Mellitus/tratamento farmacológico , Diabetes Mellitus/imunologia , Doenças do Cão/tratamento farmacológico , Cães , Ensaio de Imunoadsorção Enzimática , Feminino , Insulina/química , Insulina/uso terapêutico , Masculino , SuínosRESUMO
The cancer microenvironment plays a pivotal role in oncogenesis, containing a number of regulatory cells that attenuate the anti-neoplastic immune response. While the negative prognostic impact of regulatory T cells (Tregs) in the context of most solid tissue tumors is well established, their role in lymphoid malignancies remains unclear. T cells expressing FOXP3 and Helios were documented in the fine needle aspirates of affected lymph nodes of dogs with spontaneous multicentric B cell lymphoma (BCL), proposed to be a model for human non-Hodgkin lymphoma. Multivariable analysis revealed that the frequency of lymph node FOXP3(+) T cells was an independent negative prognostic factor, impacting both progression-free survival (hazard ratio 1.10; p = 0.01) and overall survival (hazard ratio 1.61; p = 0.01) when comparing dogs showing higher than the median FOXP3 expression with those showing the median value of FOXP3 expression or less. Taken together, these data suggest the existence of a population of Tregs operational in canine multicentric BCL that resembles thymic Tregs, which we speculate are co-opted by the tumor from the periphery. We suggest that canine multicentric BCL represents a robust large animal model of human diffuse large BCL, showing clinical, cytological and immunophenotypic similarities with the disease in man, allowing comparative studies of immunoregulatory mechanisms.
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Doenças do Cão/diagnóstico , Fatores de Transcrição Forkhead/análise , Linfoma de Células B/diagnóstico , Linfoma de Células B/veterinária , Linfócitos T Reguladores/patologia , Animais , Antígenos CD8/análise , Antígenos CD8/imunologia , Modelos Animais de Doenças , Doenças do Cão/imunologia , Doenças do Cão/patologia , Cães , Feminino , Fatores de Transcrição Forkhead/imunologia , Genes MHC da Classe II , Fator de Transcrição Ikaros/análise , Fator de Transcrição Ikaros/imunologia , Imunofenotipagem , Linfoma de Células B/imunologia , Linfoma de Células B/patologia , Masculino , Prognóstico , Linfócitos T Reguladores/imunologia , Microambiente TumoralRESUMO
Remarkably little has been published on hematological phenotypes of the domestic dog, the most polymorphic species on the planet. Information on the signalment and complete blood cell count of all dogs with normal red and white blood cell parameters judged by existing reference intervals was extracted from a veterinary database. Normal hematological profiles were available for 6046 dogs, 5447 of which also had machine platelet concentrations within the reference interval. Seventy-five pure breeds plus a mixed breed control group were represented by 10 or more dogs. All measured parameters except mean corpuscular hemoglobin concentration (MCHC) varied with age. Concentrations of white blood cells (WBCs), neutrophils, monocytes, lymphocytes, eosinophils and platelets, but not red blood cell parameters, all varied with sex. Neutering status had an impact on hemoglobin concentration, mean corpuscular hemoglobin (MCH), MCHC, and concentrations of WBCs, neutrophils, monocytes, lymphocytes and platelets. Principal component analysis of hematological data revealed 37 pure breeds with distinctive phenotypes. Furthermore, all hematological parameters except MCHC showed significant differences between specific individual breeds and the mixed breed group. Twenty-nine breeds had distinctive phenotypes when assessed in this way, of which 19 had already been identified by principal component analysis. Tentative breed-specific reference intervals were generated for breeds with a distinctive phenotype identified by comparative analysis. This study represents the first large-scale analysis of hematological phenotypes in the dog and underlines the important potential of this species in the elucidation of genetic determinants of hematological traits, triangulating phenotype, breed and genetic predisposition.