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1.
Vet Parasitol ; 271: 38-44, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31303201

RESUMO

A nucleoside triphosphate diphosphohydrolase-1 (NTPDase 1) was identified on the surface, flagellum and kinetoplast from L. infantum promastigotes by immunocytochemistry and confocal laser scanning microscopy, using immune sera that recognized specifically the B domain of NTPDase 1 and produced against synthetic peptides (LbB1LJ and LbB2LJ) derived from this domain. The polyclonal antibodies had effective antileishmanial effect, reducing significantly in vitro promastigotes growth (21-25%), an antiproliferative effect also demonstrated by immune sera produced against recombinant r-pot B domain, and two other synthetic peptides (potB1LJ and potB2LJ). In addition, using these biomolecules in ELISA technique, IgG1 and IgG2 subclasses reactivities of either healthy dogs or infected by L. infantum and classified clinically as asymptomatic, oligosymptomatic and symptomatic were tested. Analysis of distinct IgG1 and IgG2 seropositivities patterns suggested antibody subclasses binding epitopes along B domain for protection against infection, indicating this domain as a new tool for prophylactic and immunotherapeutic investigations.


Assuntos
Anticorpos Antiprotozoários/imunologia , Doenças do Cão/imunologia , Imunoglobulina G/imunologia , Leishmania infantum/enzimologia , Leishmania infantum/imunologia , Leishmaniose Visceral/veterinária , Nucleosídeo-Trifosfatase/imunologia , Animais , Anticorpos Antiprotozoários/metabolismo , Doenças do Cão/parasitologia , Cães , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/parasitologia , Domínios Proteicos/imunologia
2.
Vet Parasitol ; 271: 87-97, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31303211

RESUMO

The natural history of canine visceral leishmaniasis (CVL) has been well described, particularly with respect to the parasite load in different tissues and immunopathological changes according to the progression of clinical forms. The biomarkers evaluated in these studies provide support for the improvement of the tools used in developing vaccines against CVL. Thus, we describe the major studies using the dog model that supplies the rationale for including different biomarkers (tissue parasitism, histopathology, hematological changes, leucocytes immunophenotyping, cytokines patterns, and in vitroco-culture systems using purified T-cells subsets and macrophages infected with L. infantum) for immunogenicity and protection evaluations in phases I and II applied to pre-clinical and clinical vaccine trials against CVL. The search for biomarkers related to resistance or susceptibility has revealed a mixed cytokine profile with a prominent proinflammatory immune response as relevant for Leishmania replication at low levels as observed in asymptomatic dogs (highlighted by high levels of IFN-γ and TNF-α and decreased levels in IL-4, TGF-ß and IL-10). Furthermore, increased levels in CD4+ and CD8+ T-cell subsets, presenting intracytoplasmic proinflammatory cytokine balance, have been associated with a resistance profile against CVL. In contrast, a polyclonal B-cell expansion towards plasma cell differentiation contributes to high antibody production, which is the hallmark of symptomatic dogs associated with high susceptibility in CVL. Finally, the different studies used to analyze biomarkers have been incorporated into vaccine immunogenicity and protection evaluations. Those biomarkers identified as resistance or susceptibility markers in CVL have been used to evaluate the vaccine performance against L. infantum in a kennel trial conducted before the field trial in an area known to be endemic for visceral leishmaniasis. This rationale has been a guiding force in the testing and selection of the best vaccine candidates against CVL and provides a way for the veterinary industry to register commercial immunobiological products.


Assuntos
Biomarcadores/sangue , Doenças do Cão/sangue , Leishmaniose Visceral/veterinária , Animais , Biomarcadores/análise , Suscetibilidade a Doenças/metabolismo , Doenças do Cão/imunologia , Doenças do Cão/parasitologia , Cães , Leishmaniose Visceral/sangue , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/parasitologia , Vacinas Protozoárias/imunologia
3.
BMC Vet Res ; 15(1): 185, 2019 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-31164117

RESUMO

BACKGROUND: Ex vivo whole blood stimulation assays (WBSA) have been used to characterize the cytokine response to diet in cats. The present study aimed to use this assay to determine the cytokine response to diets being fed at the time of diagnosis to dogs with chronic enteropathy (CE) and to compare this to a control group of dogs presented for non-gastrointestinal (GI) causes. RESULTS: Dogs with chronic GI signs and dogs presented for non-GI causes were prospectively recruited. For each case, residual blood following diagnostic sampling was placed into heparin. WBSAs were performed using crude extracts of the diet currently being fed and provided by the owner. Supernatants were collected and analyzed for tumor necrosis factor (TNF)-alpha, interleukin (IL)-10 and IL-4 using an enzyme-linked immunosorbent assay. The case group consisted of 22 dogs with CE diagnosed on histopathology of GI biopsy and 9 with suspected CE. The non-GI group consisted of 18 dogs. Of the diets being fed at or prior to diagnosis, hydrolyzed protein diets elicited significantly lower IL-10 and TNF-alpha concentrations compared to commercial intact protein diets in dogs with confirmed or suspected CE (P-value 0.004 and < 0.001, respectively). Six out of 15 dogs with detectable IL-4 concentrations in the confirmed CE group had IL-4 to IL-10 ratios that exceeded the 95% confidence interval (CI) of the mean of the non-GI group (non-GI: 95% CI of IL-4:IL-10 = 0.64-2.71; confirmed CE: IL-4:IL-10 in 6 dogs = mean 22.40, range 2.77-89.11). CONCLUSIONS: Hydrolyzed protein diets elicited a significantly reduced cytokine response when incubated with patient whole blood ex vivo compared to commercial intact protein diets in dogs with CE. The IL-4 to IL-10 ratio as a marker of dietary responsiveness warrants further investigation, together with assessment of the cytokine response to diet at the intestinal mucosal surface.


Assuntos
Citocinas/sangue , Dieta/veterinária , Doenças do Cão/imunologia , Enteropatias/veterinária , Animais , Doença Crônica , Proteínas na Dieta/farmacologia , Cães , Feminino , Enteropatias/imunologia , Masculino , Estudos Prospectivos
4.
Vet Immunol Immunopathol ; 212: 43-49, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31213251

RESUMO

Canine atopic dermatitis (AD) is a chronic, inflammatory and pruritic allergic skin disease in dogs. House dust mites such as Dermatophagoides farinae are one of the known causative agents for the induction of canine AD worldwide. D. farinae protein Der f 2 is known as an important allergen involved in canine AD and recently, Zen-1 has also been identified as an allergenic protein. There is limited information on the prevalence and role of allergen sensitization to crude D. farinae extract (CDF), Der f 2 and Zen-1 among dogs diagnosed with AD in Malaysia. The aim of this study was to determine the proportion of CDF-, Der f 2- and Zen-1-specific reactive sera among dogs diagnosed with AD in Malaysia using an enzyme-linked immunosorbent assay (ELISA). Serum samples were collected from dogs diagnosed with AD from several veterinary clinics in Malaysia. The canine case records were retrieved and information on signalment, dermatological and non-dermatological histories, clinical presentation, food allergies, and exclusion of ectoparasitic, microbial and fungal skin infections were obtained through a survey form. All serum samples were evaluated to quantify the CDF-, Der f 2- and Zen-1-specific immunoglobulin E (IgE) levels. A total of 24.6%, 48.4% and 29.8% of dogs diagnosed with AD were positive for CDF-, Der f 2- and Zen-1-specific IgE, respectively. These results suggest that CDF-, Der f 2- and Zen-1 are important allergens that can contribute to AD in dogs in Malaysia, and serological testing can be performed to provide additional treatment options involving specific immunotherapies.


Assuntos
Antígenos de Dermatophagoides/imunologia , Proteínas de Artrópodes/imunologia , Dermatite Atópica/veterinária , Doenças do Cão/imunologia , Imunoglobulina E/sangue , Alérgenos/imunologia , Animais , Antígenos de Dermatophagoides/sangue , Proteínas de Artrópodes/sangue , Dermatite Atópica/imunologia , Dermatite Atópica/parasitologia , Dermatophagoides farinae , Doenças do Cão/parasitologia , Cães , Ensaio de Imunoadsorção Enzimática , Hospitais Veterinários , Malásia , Animais de Estimação/imunologia
5.
Vet Parasitol ; 269: 34-41, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31079826

RESUMO

The outcome of infection with Leishmania infantum in dogs is variable, which is thought to be due to the nature of the immune response mounted by the host. As a consequence, the clinical signs and severity of canine leishmaniosis vary between individual dogs. Host immunogenetic factors might play an important role in determining the outcome of infection. The aim of this study was to examine polymorphisms in innate and adaptive immune response genes, to determine whether any of these were associated with susceptibility or resistance to L. infantum infection. Genomic DNA was obtained from two groups: pet dogs in endemic regions of Europe and a group of Beagles exposed to sand fly infection as part of a vaccine study. Genotyping was performed using a SNP (single nucleotide polymorphism) array for selected immune response genes. The first part of the study compared 62 clinical cases with 101 clinically unaffected dogs that were seronegative for Leishmania antibodies. One SNP in the CIITA gene demonstrated a significantly higher minor allele frequency in the case group, compared with the control group at the individual SNP level after permutation, but was not significant after correction for multiple testing. The second part of the study examined 48 Beagle dogs exposed to L. infantum over two transmission seasons. Twenty-seven dogs with a resistant phenotype (no evidence of clinical disease, seronegative at the end of the study period, negative on lymph node culture and only transiently PCR positive in bone marrow) were compared with 21 dogs demonstrating a susceptible phenotype (clinical disease, seropositive, positive lymph node culture and consistently PCR positive in bone marrow). Three SNPs in TLR3, two SNPs in PTPN22 and one SNP in TLR4 and IL1A were associated with the susceptible phenotype in the Beagle group at the individual SNP level after permutation analysis, but were not significant after correction for multiple testing. Further validation of these SNPs is required in a larger cohort of dogs, ideally with extreme phenotypes to confirm an association with the outcome of L. infantum infection.


Assuntos
Resistência à Doença/genética , Doenças do Cão/imunologia , Leishmania infantum/imunologia , Leishmaniose/veterinária , Polimorfismo de Nucleotídeo Único/genética , Psychodidae/parasitologia , Imunidade Adaptativa/genética , Animais , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães , Doenças Endêmicas/veterinária , Europa (Continente)/epidemiologia , Imunidade Inata/genética , Leishmania infantum/genética , Leishmaniose/epidemiologia , Leishmaniose/imunologia , Leishmaniose/parasitologia , Reação em Cadeia da Polimerase/veterinária
6.
Vet Immunol Immunopathol ; 211: 64-74, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31084897

RESUMO

S100A12 and S100A8/A9 (calprotectin) are released from activated mononuclear cells and belong to the group of damage associated molecular patterns. Fecal S100A12 and S100A8/A9 concentrations have been suggested as biomarkers of intestinal inflammation in dogs with chronic inflammatory enteropathies (CIE). However, the mucosal cellular infiltrate in dogs with CIE is primarily lymphocytic-plasmacytic. Whether fecal S100A12 and S100A8/A9 levels reflect the number and/or activity of intestinal mucosal mononuclear cells, or whether these proteins are also produced by other cells has not been investigated. Thus, the aim of this study was to evaluate intestinal mucosal S100A12 and S100A8/A9 positivity and a potential relationship with the respective protein concentrations in serum and fecal samples in dogs with CIE. Serum (single sample), fecal samples (from 3 consecutive days), and gastrointestinal tissue biopsies (i.e., stomach, duodenum, ileum, and colon) were evaluated from 21 dogs with CIE. Serum and fecal S100A12 and S100A8/A9 concentrations were measured by analytically validated in-house ELISAs. Tissue biopsies underwent routine histopathology and immunohistochemical evaluation for S100A12 and S100A8/A9 positivity (S100A12+ and S100A8/A9+, each recorded as positive cells/mm2). S100A12+ and S100A8/A9+ cells were identified in all segments of the gastrointestinal tract, but were predominantly localized in the lamina propria (LP). Duodenal LP S100A12 positivity correlated statistically significantly with that in the stomach and ileum (ρ = 0.66 and 0.69, both p < 0.01), but was inversely correlated with the severity of macrophage infiltration in the duodenum (ρ=-0.47, p = 0.042). Ileal LP S100A8/A9 positivity correlated positively with the extent of ileal neutrophil and macrophage infiltration (ρ=0.61, p = 0.047). Fecal S100A12 concentrations strongly correlated with the number of S100A12+ cells along the entire gastrointestinal tract (ρ = 0.76, p = 0.028), whereas serum S100A12 concentrations were inversely correlated to colonic S100A12+ cell counts (ρ=-0.50, p = 0.043). Mucosal S100A8/A9+ cell counts were not associated with the corresponding fecal or serum S100A8/A9 concentrations. These results suggest that the intestinal mucosa in dogs with CIE contains an increased number of activated (pro-inflammatory) phagocytes expressing and secreting the S100A12 protein, but the macrophage population seen on routine histopathology is predominantly mature (anti-inflammatory) with a reduced or absent expression of S100A12 and a normal or increased expression of S100A8/A9. However, the distribution of intestinal S100A8/A9 expression requires further study.


Assuntos
Doenças do Cão/imunologia , Doenças Inflamatórias Intestinais/veterinária , Mucosa Intestinal/metabolismo , Complexo Antígeno L1 Leucocitário/metabolismo , Proteína S100A12/metabolismo , Animais , Biomarcadores/análise , Biomarcadores/sangue , Doenças do Cão/sangue , Cães , Fezes/química , Feminino , Doenças Inflamatórias Intestinais/sangue , Doenças Inflamatórias Intestinais/imunologia , Complexo Antígeno L1 Leucocitário/análise , Complexo Antígeno L1 Leucocitário/sangue , Masculino , Neutrófilos/imunologia , Proteína S100A12/análise , Proteína S100A12/sangue
7.
Am J Vet Res ; 80(6): 572-577, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31140843

RESUMO

OBJECTIVE: To investigate luteinizing hormone (LH) receptor expression in canine nonneoplastic and neoplastic lymph nodes, circulating nonneoplastic lymphocytes, and T-cell lymphoma (TCL) cell lines. SAMPLE: Formalin-fixed, paraffin-embedded lymph nodes (5 neoplastic and 3 nonneoplastic) from 6 dogs, circulating lymphocytes from venous blood specimens obtained from 12 healthy dogs, and 3 TCL cell lines derived from 3 dogs with primary lymphoma. PROCEDURES: Lymph node specimens were immunohistochemically stained for determination of LH receptor expression. Circulating nonneoplastic lymphocytes and TCL cell lines were evaluated for LH receptor expression by use of flow cytometry; circulating lymphocytes were also immunophenotyped. The mean percentage of cells positive for LH receptors was determined for each type of specimen. For the healthy dogs, percentages of circulating B and T lymphocytes that expressed LH receptors were assessed on the basis of sex and reproductive status. RESULTS: The mean percentage of LH receptor-positive cells in canine neoplastic and nonneoplastic lymph nodes was 12.4% and 4.1%, respectively. For the healthy dogs, the mean percentage of circulating LH receptor-positive T lymphocytes was significantly higher in gonadectomized dogs (16.6%) than in sexually intact dogs (10.5%); the percentages of circulating LH receptor-positive B lymphocytes did not significantly differ by reproductive status. Among the 3 canine TCL cell lines, LH receptor expression ranged from 10% to 45%. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, LH receptor expression by canine neoplastic and nonneoplastic lymphocytes was detected. Research into the effects of downregulation of LH receptor activation in dogs with lymphoma is warranted.


Assuntos
Doenças do Cão/metabolismo , Linfócitos/metabolismo , Linfoma/veterinária , Receptores do LH/biossíntese , Linfócitos T/metabolismo , Animais , Linhagem Celular Tumoral , Doenças do Cão/imunologia , Doenças do Cão/patologia , Cães , Feminino , Citometria de Fluxo/veterinária , Linfonodos/metabolismo , Linfoma/imunologia , Linfoma/metabolismo , Masculino
8.
Int J Mol Sci ; 20(10)2019 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-31091785

RESUMO

Growing scientific evidence has unveiled increased incidences of obesity in domestic animals and its influence on a plethora of associated disorders. Leptin, an adipokine regulating body fat mass, represents a key molecule in obesity, able to modulate immune responses and foster chronic inflammatory response in peripheral tissues. High levels of cytokines and inflammatory markers suggest an association between inflammatory state and obesity in dogs, highlighting the parallelism with humans. Canine obesity is a relevant disease always accompanied with several health conditions such as inflammation, immune-dysregulation, insulin resistance, pancreatitis, orthopaedic disorders, cardiovascular disease, and neoplasia. However, leptin involvement in many disease processes in veterinary medicine is poorly understood. Moreover, hyperleptinemia as well as leptin resistance occur with cardiac dysfunction as a consequence of altered cardiac mitochondrial metabolism in obese dogs. Similarly, leptin dysregulation seems to be involved in the pancreatitis pathophysiology. This review aims to examine literature concerning leptin and immunological status in obese dogs, in particular for the aspects related to obesity-associated diseases.


Assuntos
Imunidade Adaptativa , Doenças do Cão/imunologia , Imunidade Inata , Leptina/genética , Obesidade/veterinária , Animais , Doenças do Cão/etiologia , Cães , Leptina/metabolismo , Obesidade/etiologia , Obesidade/imunologia
9.
Vet Microbiol ; 231: 177-182, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30955806

RESUMO

Canine parvovirus (CPV) is one of the most important cause of mortality in young dogs and no specific treatment exists. Since prolonged leukopenia greatly increases the risk of death in infected pups, strategies to counteract this decline were investigated. The outcomes of CPV naturally infected pups treated with the recombinant canine granulocyte-colony stimulating factor (rcG-CSF), in combination with the routine therapy, were compared with similarly-managed infected pups not treated with rcG-CSF. A non-randomized prospective clinical trial was performed on 62 CPV infected pups with WBC counts <3000 cells/µL and two different groups were selected based on a non-randomized approach. Group A dogs (31/62) received 5 µg/Kg of rcG-CSF daily from the hospitalization day until WBC reached the reference range (3-5 days) and group B (31/62) received 1 ml of placebo injection. All dogs in group A recovered, while five dogs in group B died. The rcG-CSF treatment demonstrated a statistically significant effect on WBC counts (p < 0.0001) and, surprisingly, also on lymphocytes and monocytes counts (p < 0.0001). There was no significant effect of treatment on neutrophil count (p = 0.5502). Although lymphocytes and monocytes are not a specific target for rcG-CSF, our study highlights that rcG-CSF is able to improve haematological parameters compared to untreated dogs and a clear increase in their number was detected, as previously described for humans treated with the homologous molecule.


Assuntos
Doenças do Cão/prevenção & controle , Fator Estimulador de Colônias de Granulócitos/imunologia , Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Contagem de Leucócitos/veterinária , Infecções por Parvoviridae/veterinária , Animais , Doenças do Cão/imunologia , Doenças do Cão/virologia , Cães , Feminino , Masculino , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/prevenção & controle , Parvovirus Canino , Estudos Prospectivos , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/uso terapêutico
10.
J Vet Intern Med ; 33(3): 1392-1402, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30939225

RESUMO

BACKGROUND: Polymerase chain reaction for antigen receptor rearrangement (PARR) is a molecular diagnostic tool used for discrimination of lymphoid malignancies in dogs from benign processes. Assay variations have been described and are commercially available, but performance metrics are not uniformly reported. OBJECTIVES: To describe performance (accuracy, sensitivity, specificity) and rigorous benchmarking of a PARR protocol (ePARR) in clinically relevant samples. ANIMALS: One hundred eighty-one client-owned dogs. METHODS: Lymphoma and benign tissues representative of the clinical spectrum with gold standard histopathologic and immunohistochemical diagnoses were collected. Assay development and benchmarking were performed on fresh frozen (FF) tissue, formalin-fixed paraffin-embedded (FFPE) tissue, flow cytometry pellets, and air-dried fine-needle aspirates (FNA). Assay performance was determined for FFPE from 56 dogs (18 B-cell lymphoma, 24 T-cell lymphoma, and 14 non-lymphoma), 80 frozen flow cytometry pellets (66 B-cell lymphoma, 14 T-cell lymphoma, 0 non-lymphoma), and 41 air-dried FNA slides (23 lymphoma, 18 non-lymphoma). RESULTS: For discrimination of lymphoma versus non-lymphoma, ePARR had 92% and 92% sensitivity and specificity on FFPE with 92% accuracy, 85% sensitivity from flow cytometry pellets (non-lymphoma was not evaluated to calculate specificity) with 85% accuracy, and 100% and 100% sensitivity and specificity for FNA with 100% accuracy. Stringent quality control criteria decreased assay success rate without significant performance improvement. Performance metrics were lower in most cases for discrimination of B- or T-cell versus non-B- or non-T-cell samples than for lymphoma versus non-lymphoma. CONCLUSIONS AND CLINICAL IMPORTANCE: These benchmarking data facilitate effective interpretation and application of PARR assays in multiple sample types.


Assuntos
Doenças do Cão/genética , Doenças do Cão/imunologia , Rearranjo Gênico , Linfoma/veterinária , Reação em Cadeia da Polimerase/veterinária , Animais , Benchmarking , Doenças do Cão/diagnóstico , Cães , Imunofenotipagem/veterinária , Linfoma/genética , Linfoma/imunologia , Linfoma de Células B/genética , Linfoma de Células B/imunologia , Linfoma de Células B/veterinária , Linfoma de Células T/genética , Linfoma de Células T/imunologia , Linfoma de Células T/veterinária , Reação em Cadeia da Polimerase/métodos , Estudos Prospectivos , Receptores de Antígenos/genética
11.
Anticancer Res ; 39(4): 1699-1703, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30952708

RESUMO

BACKGROUND/AIM: Previous work in rodent models showed that an autologous tissue vaccine is both a safe and effective approach for treating cancer; however, as a translational step, safety must first be evaluated in a more clinically-relevant model. MATERIALS AND METHODS: An autologous immunotherapy produced from resected tumors, was evaluated in a clinically-relevant canine model to assess safety. Ninety-three dogs with spontaneously occurring tumors received vaccination with inactivated autologous tumor tissue combined with an adjuvant of particulate porcine small intestinal submucosa extracellular matrix (SIS-ECM). Patients were followed to assess the occurrence of adverse events, overall survival, and tumor recurrence and/or metastasis. RESULTS: A small number (12%) of patients experienced limited, mild pyrexia, injection site swelling, or lethargy, all resolving without clinical intervention. CONCLUSION: Autologous whole cell cancer immunotherapy can be used safely in the canine model of cancer and represents a safe approach for the treatment for cancer.


Assuntos
Antígenos de Neoplasias/administração & dosagem , Vacinas Anticâncer/administração & dosagem , Doenças do Cão/terapia , Imunoterapia/veterinária , Neoplasias/veterinária , Adjuvantes Imunológicos , Animais , Antígenos de Neoplasias/imunologia , Antígenos de Neoplasias/toxicidade , Vacinas Anticâncer/imunologia , Vacinas Anticâncer/toxicidade , Doenças do Cão/diagnóstico , Doenças do Cão/imunologia , Doenças do Cão/cirurgia , Cães , Feminino , Imunoterapia/efeitos adversos , Mucosa Intestinal/imunologia , Intestino Delgado/imunologia , Masculino , Margens de Excisão , Neoplasia Residual , Neoplasias/diagnóstico , Neoplasias/imunologia , Neoplasias/terapia , Sus scrofa
12.
Vet J ; 245: 1-6, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30819419

RESUMO

An elimination diet (ED) followed by re-challenge has been the reference standard to diagnose adverse food reactions (AFR) in dogs, but can be challenging to conduct. This study investigated the accuracy of a saliva-based test for food-specific IgA and IgM and an ELISA serum test for food-specific IgE. Three groups of dogs were tested. Group 1 (n=11) included dogs with previously diagnosed and controlled AFR; group 2 (n=15) comprised dogs with allergic dermatitis at the beginning of their ED; and group 3 (n=16) was composed of clinically healthy research dogs. Saliva samples were collected from all groups and blood samples from group 1 and group 3. The results of clinical re-challenges with individual food components were compared with the test results. Specificity, sensitivity, positive and negative predictive values and likelihood ratios were determined. Forty-one dogs completed the study; one dog was lost to follow up. There was a total of 163 re-challenges. Sensitivity, positive predictive value and likelihood ratio, specificity, negative predictive value and likelihood ratios were unsatisfactory for both tests in most instances, except for IgM testing in group 2, which had moderate specificity. There was no clear difference in the number of positive reactions between the allergic dogs and healthy dogs from a research population. Based on these results, the saliva test for food specific IgA and IgM and the ELISA serum test for food specific IgE were not reliable to diagnose adverse food reactions in dogs. Until more data are available, elimination diets remain the reference standard in the diagnosis of this disease.


Assuntos
Anticorpos/análise , Dieta/veterinária , Doenças do Cão/imunologia , Hipersensibilidade Alimentar/veterinária , Saliva/imunologia , Alérgenos , Animais , Dieta/efeitos adversos , Cães/imunologia , Hipersensibilidade Alimentar/imunologia , Imunoglobulina A/análise , Imunoglobulina E/sangue , Imunoglobulina M/análise , Sensibilidade e Especificidade
13.
Vet Dermatol ; 30(3): 195-e61, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30908739

RESUMO

BACKGROUND: Tests for allergen-specific immunoglobulin E (IgE) are used to select allergens for immunotherapy in atopic dogs. Antibodies against cross-reactive carbohydrate determinants (anti-CCD IgE) have been identified in serum samples of atopic dogs. Their presence in humans is a known cause of clinically irrelevant polysensitization to plant allergens. OBJECTIVES: To compare the results of an intradermal test (IDT) and a serum test for allergen-specific IgE, with and without blocking anti-CCD IgE, before testing in dogs. ANIMALS: Thirty-one privately owned dogs with atopic dermatitis. METHODS AND MATERIALS: Dogs were prospectively skin tested and their serum samples were analysed for anti-CCD IgE. An Fc-ε receptor-based serum test for allergen-specific IgE was performed with and without blocking anti-CCD IgE. RESULTS: In dogs with negative anti-CCD IgE samples, the agreement between the results of the serum test and the IDT was substantial (κ = 0.71). Dogs with positive anti-CCD IgE samples (38.7%) showed no agreement between serum and skin testing (κ = -0.35), blocking anti-CCD IgE in those samples resulted in a fair agreement (κ = 0.43). Anti-CCD IgE positive sera had multiple positive results for grass and weed allergens, and blocking decreased them markedly. CONCLUSION AND CLINICAL IMPORTANCE: Intradermal testing agreed best with serum testing in dogs with no detectable anti-CCD IgE. Sera containing anti-CCD IgE had no agreement with IDT. Test agreement was improved by blocking the anti-CCD IgE. Apparent serum test polysensitization to plant allergens was associated with anti-CCD IgE.


Assuntos
Alérgenos/imunologia , Carboidratos/imunologia , Dermatite Atópica/veterinária , Imunoglobulina E/sangue , Testes Intradérmicos/veterinária , Testes Cutâneos/veterinária , Animais , Reações Cruzadas , Dermatite Atópica/diagnóstico , Doenças do Cão/diagnóstico , Doenças do Cão/imunologia , Cães , Ensaio de Imunoadsorção Enzimática/veterinária , Testes Intradérmicos/métodos , Estudos Prospectivos , Testes Sorológicos/métodos , Testes Sorológicos/veterinária
14.
Parasit Vectors ; 12(1): 119, 2019 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-30909975

RESUMO

BACKGROUND: Canine leishmaniosis (CanL) due to Leishmania infantum is characterized by the development of both cellular and humoral immune responses. The dysfunction of T cell-mediated immunity leads to a lack of proliferation of T cells in response to Leishmania antigens with the consequence of parasite dissemination that seems to be related to a T cell exhaustion mediated by regulatory B cells expressing immunoglobulin D (IgD). The aim of this study was to determine and compare the total serum IgD in dogs with clinical leishmaniosis and in clinically healthy dogs. RESULTS: A total of 147 dog sera were studied. All dogs were tested for L. infantum-specific antibodies by quantitative ELISA. Interferon-gamma (IFN-γ) production was also determined by sandwich ELISA after blood stimulation with L. infantum soluble antigen (LSA) or concanavalin A (ConA). The quantification of total IgD was performed using a human IgD sandwich ELISA quantification set. Dogs were classified in three different groups. Group 1 included 40 clinically healthy non-infected dogs, all serologically negative to L. infantum-specific antibodies and non-producers of IFN-γ upon LSA stimulation. Group 2 included 63 clinically healthy infected dogs that were LSA IFN-γ producers (n = 61) and/or IFN-γ non-producers (n = 2) as well as negative to medium seropositive to L. infantum antigen. Finally, Group 3 included 44 dogs with clinical leishmaniosis (IFN-γ producers, n = 23; and IFN-γ non-producers, n = 21) that were negative to highly positive to L. infantum-specific antibodies. No significant differences were observed when the total IgD concentration was compared within groups. Additionally, total IgD of sick IFN-γ producers and IFN-γ non-producers was not significantly different. Finally, total IgD concentration was not statistically related to demographic parameters such as age, sex and breed. CONCLUSIONS: The results of this study demonstrated that there were no differences between groups in total serum IgD. Total serum IgD does not appear to be a marker of disease in CanL.


Assuntos
Anticorpos Antiprotozoários/sangue , Doenças do Cão/parasitologia , Imunoglobulina D/imunologia , Leishmania infantum/imunologia , Leishmaniose Visceral/veterinária , Animais , Anticorpos Antiprotozoários/imunologia , Doenças do Cão/sangue , Doenças do Cão/imunologia , Cães , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Imunoglobulina D/sangue , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/parasitologia , Masculino , Estudos Retrospectivos
15.
Top Companion Anim Med ; 34: 22-29, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30808493

RESUMO

Immune-mediated inflammation is responsible for about 25% of central nervous system disease in dogs. The disease can affect all ages and breeds, but young to middle-aged small breed dogs are over-represented for most forms. Diagnosis consists of advanced imaging (MRI), cerebrospinal fluid analysis, and infectious disease testing, but biopsy is required for definitive diagnosis and classification of the disease into one of the many subtypes. Treatment consists of immunosuppressive medication with the goal being to control and/or improve clinical signs. Current literature shows that prognosis is variable.


Assuntos
Doenças do Sistema Nervoso Central/veterinária , Doenças do Cão/imunologia , Inflamação/veterinária , Animais , Biópsia/veterinária , Doenças do Sistema Nervoso Central/diagnóstico , Doenças do Sistema Nervoso Central/tratamento farmacológico , Doenças do Sistema Nervoso Central/imunologia , Doenças do Cão/diagnóstico , Doenças do Cão/tratamento farmacológico , Cães , Feminino , Imunossupressores/uso terapêutico , Inflamação/patologia , Imagem por Ressonância Magnética/veterinária , Masculino , Prognóstico
16.
Artigo em Inglês | MEDLINE | ID: mdl-30711049

RESUMO

Brucella canis is a small intracellular Gram-negative bacterium that frequently leads to chronic infections highly resistant to antibiotic therapy in dogs. Also, it causes mild human brucellosis compared to other zoonotic Brucella spp. Herein we characterize the cellular immune response elicited by B. canis by analysing human and canine CD4+ T cells after stimulation with autologous monocyte-derived dendritic cells (MoDCs). Human and canine B. canis-primed MoDCs stimulated autologous CD4+ T cells; however, a Th1 response was triggered by human MoDCs, whereas canine MoDCs induced Th1/Th17 responses, with increased CD4+ T cells producing IFN-γ and IL-17A simultaneously. Each pattern of cellular response may contribute to host susceptibility, helping to understand the differences in B. canis virulence between these two hosts. In addition, other aspects of canine immunology are unveiled by highlighting the participation of IL-17A-producing canine MoDCs and CD4+ T cells producing IFN-γ and IL-17A.


Assuntos
Brucella canis/imunologia , Linfócitos T CD4-Positivos/fisiologia , Citocinas/biossíntese , Células Dendríticas/imunologia , Células Th1/fisiologia , Células Th17/fisiologia , Animais , Biomarcadores , Brucelose/veterinária , Comunicação Celular/imunologia , Células Dendríticas/metabolismo , Doenças do Cão/imunologia , Doenças do Cão/metabolismo , Doenças do Cão/microbiologia , Cães , Humanos , Imunofenotipagem , Ativação Linfocitária
17.
Artigo em Inglês | MEDLINE | ID: mdl-30711051

RESUMO

PD-1 is a negative costimulator of chronic infectious diseases In this study, we investigated the expression of PD-1 and its ligands in the spleen of dogs with visceral leishmaniasis and lymphoproliferative response to soluble antigen, in lymph node cells in the presence or absence of antibodies blocking PD-1 and its ligands. Our results showed expression of PD-1 and its ligands is higher after L. infantum infection and in the spleen of infected dogs, PD-1 blockage was able to restore the antigen-dependent lymphoproliferative response and regulated production of the cytokines IL-4 and IL-10 and NO production. We concluded that L. infantum infection modulates PD-1 and its ligands expression in canine VL and that blockage of PD-1 restores the immune response. Thus, blockage of PD-1 is a target for therapeutic drug development.


Assuntos
Antígeno B7-H1/metabolismo , Doenças do Cão/imunologia , Doenças do Cão/metabolismo , Imunidade Celular , Leishmaniose Visceral/veterinária , Receptor de Morte Celular Programada 1/metabolismo , Animais , Antígeno B7-H1/genética , Citocinas/metabolismo , Doenças do Cão/parasitologia , Cães , Feminino , Expressão Gênica , Imuno-Histoquímica , Imunofenotipagem , Leucócitos/imunologia , Leucócitos/metabolismo , Linfócitos/imunologia , Linfócitos/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Carga Parasitária , Receptor de Morte Celular Programada 1/genética , Baço/imunologia , Baço/metabolismo
18.
J Vet Intern Med ; 33(2): 764-775, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30666722

RESUMO

BACKGROUND: T-zone lymphoma (TZL), an indolent disease in older dogs, comprises approximately 12% of lymphomas in dogs. TZL cells exhibit an activated phenotype, indicating the disease may be antigen-driven. Prior research found that asymptomatic aged Golden Retrievers (GLDRs) commonly have populations of T-zone-like cells (phenotypically identical to TZL) of undetermined significance (TZUS). OBJECTIVE: To evaluate associations of inflammatory conditions, TZL and TZUS, using a case-control study of GLDRs. ANIMALS: TZL cases (n = 140), flow cytometrically diagnosed, were identified through Colorado State University's Clinical Immunology Laboratory. Non-TZL dogs, recruited through either a database of owners interested in research participation or the submitting clinics of TZL cases, were subsequently flow cytometrically classified as TZUS (n = 221) or control (n = 147). METHODS: Health history, signalment, environmental, and lifestyle factors were obtained from owner-completed questionnaires. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were estimated using multivariable logistic regression, obtaining separate estimates for TZL and TZUS (versus controls). RESULTS: Hypothyroidism (OR, 0.3; 95% CI, 0.1-0.7), omega-3 supplementation (OR, 0.3; 95% CI, 0.1-0.6), and mange (OR, 5.5; 95% CI, 1.4-21.1) were significantly associated with TZL. Gastrointestinal disease (OR, 2.4; 95% CI, 0.98-5.8) had nonsignificantly increased TZL odds. Two shared associations for TZL and TZUS were identified: bladder infection or calculi (TZL OR, 3.5; 95% CI, 0.96-12.7; TZUS OR, 5.1; 95% CI, 1.9-13.7) and eye disease (TZL OR, 2.3; 95% CI, 0.97-5.2; TZUS OR, 1.9; 95% CI, 0.99-3.8). CONCLUSIONS AND CLINICAL IMPORTANCE: These findings may elucidate pathways involved in TZUS risk and progression from TZUS to TZL. Further investigation into the protective association of omega-3 supplements is warranted.


Assuntos
Doenças do Cão/imunologia , Linfoma de Células T/veterinária , Fatores Etários , Animais , Estudos de Casos e Controles , Suplementos Nutricionais , Doenças do Cão/epidemiologia , Cães , Ácidos Graxos Ômega-3 , Feminino , Citometria de Fluxo/veterinária , Hipotireoidismo/veterinária , Linfoma de Células T/epidemiologia , Linfoma de Células T/imunologia , Masculino , Infestações por Ácaros/veterinária , Linfócitos T , Cálculos da Bexiga Urinária/veterinária , Infecções Urinárias/veterinária
19.
Talanta ; 195: 327-332, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-30625550

RESUMO

Leishmaniasis is a disease with high impact on public health in many countries. Visceral leishmaniasis (VL) is a vectorial zoonosis, with dogs as primary reservoirs in the domestic environment. VL presents high morbidity, mortality and importance in epidemiology in the American continent. In the present study, the first label-free electrochemical impedance immunosensor using screen-printed electrodes (SPEs) for the detection of anti-Leishmania infantum antibodies was developed. The soluble antigens of L. infantum were immobilized on an SPE by a 3-mercaptopropionic acid monolayer. Electrochemical impedance spectroscopy (EIS) was used for detecting bimolecular interactions occurring at the electrode surface. The addition of real samples consisting of canine and human sera positive and negative for VL presented high sensitivity and selectivity through EIS. Based on the results, a sensitive, specific, rapid and simple immunosensor was developed successfully with potential application for the serological diagnosis of leishmaniasis disease.


Assuntos
Anticorpos Antiprotozoários/sangue , Técnicas Biossensoriais , Doenças do Cão/diagnóstico , Leishmania infantum/imunologia , Leishmaniose Visceral/diagnóstico , Animais , Anticorpos Antiprotozoários/imunologia , Antígenos de Protozoários/química , Antígenos de Protozoários/imunologia , Espectroscopia Dielétrica , Doenças do Cão/sangue , Doenças do Cão/imunologia , Cães , Eletrodos , Ouro/química , Humanos , Imunoensaio , Leishmaniose Visceral/sangue , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/veterinária
20.
Vet Pathol ; 56(3): 350-357, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30636524

RESUMO

Marginal zone lymphoma (MZL) and mantle cell lymphoma (MCL) belong to a subgroup of indolent B-cell lymphomas most commonly reported in the canine spleen. The goal of this study was to characterize the immunophenotype of splenic MZL and MCL in comparison to their human counterparts. Ten MCLs and 28 MZLs were selected based on morphology. A tissue microarray was generated, and expression of CD3, CD5, CD10, CD45, CD20, CD79a, Pax-5, Bcl-2, Bcl-6, cyclin D1, cyclin D3, MCL-1, MUM-1, and Sox-11 was evaluated. Neoplastic cells in all MCLs and MZLs were positive for CD5, CD20, CD45, CD79a, and BCL2 and negative for CD3, CD10, Bcl-6, cyclin D1, and cyclin D3. Positive labeling for Pax-5 was detected in 8 of 10 MCLs and 26 of 28 MZLs. Positive labeling for MUM-1 was detected in 3 of 10 MCLs, and 27 of 28 MZLs were positive for MUM-1. No MCLs but 8 of 24 MZLs were positive for MCL-1. Canine splenic MZL and MCL have a similar immunophenotype as their human counterparts. However, human splenic MCL overexpresses cyclin D1 due to a translocation. A similar genetic alteration has not been reported in dogs. In addition, in contrast to human MZL, canine splenic MZL generally expresses CD5. Following identification of B vs T cells with CD20 and CD3, a panel composed of BCL-2, Bcl-6, MUM-1, and MCL-1 combined with the histomorphological pattern can be used to accurately diagnose MZL and MCL in dogs. Expression of Bcl-2 and lack of MCL-1 expression in MCL may suggest a therapeutic benefit of BCL-2 inhibitors in canine MCL.


Assuntos
Doenças do Cão/patologia , Imunofenotipagem/veterinária , Linfoma de Células B/veterinária , Linfoma Folicular/veterinária , Neoplasias Esplênicas/veterinária , Animais , Antígenos de Diferenciação de Linfócitos T/imunologia , Doenças do Cão/imunologia , Cães , Linfoma de Células B/imunologia , Linfoma de Células B/patologia , Linfoma de Zona Marginal Tipo Células B/imunologia , Linfoma de Zona Marginal Tipo Células B/patologia , Linfoma de Zona Marginal Tipo Células B/veterinária , Linfoma Folicular/imunologia , Linfoma Folicular/patologia , Linfoma de Célula do Manto/imunologia , Linfoma de Célula do Manto/patologia , Linfoma de Célula do Manto/veterinária , Estudos Retrospectivos , Baço/imunologia , Baço/patologia , Neoplasias Esplênicas/imunologia , Neoplasias Esplênicas/patologia
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