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1.
Vet Res ; 51(1): 110, 2020 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-32883344

RESUMO

Canine parvovirus (CPV) can cause acute and highly contagious bloody enteritis in dog. To obtain antibodies against CPV, hens were immunized with virus-like particles (VLP) of CPV-VP2. The IgY single chain fragment variables (scFv) were generated by T7 phage display system and expressed in E. coli system. The titer of the primary scFv library reached to 1.5 × 106 pfu/mL, and 95% of the phages contained the target fragments. The CPV-VLP and CPV-VP2 protein showed similar reaction values to the purified scFv in the ELISA test, and the results of ELISA analysis using IgY-scFv toward CPV clinical samples were consistent with commercial immunochromatographic assay (ICA) and PCR detection, the scFv did not show cross reactivity with canine distemper virus (CDV) and canine coronavirus (CCV). IgY-scFv was successfully expressed in CRFK cells, and in the virus suppression assay, 55% of CPV infections were eliminated within 24 h. Docking results demonstrated that the number of amino acids of the binding sides between scFv and VP2 were AA37 and AA40, respectively. This study revealed the feasibility of a novel functional antibody fragment development strategy by generating diversified avian IgY-scFv libraries towards the pathogenic target of interest for both detection and therapeutic purposes in veterinary medicine.


Assuntos
Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Proteínas do Capsídeo/imunologia , Doenças do Cão/virologia , Imunoglobulinas/imunologia , Infecções por Parvoviridae/veterinária , Parvovirus Canino/imunologia , Anticorpos de Cadeia Única/imunologia , Animais , Galinhas/imunologia , Doenças do Cão/diagnóstico , Doenças do Cão/imunologia , Cães , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Simulação de Acoplamento Molecular , Infecções por Parvoviridae/diagnóstico , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/virologia , Anticorpos de Cadeia Única/genética
2.
PLoS One ; 15(8): e0238371, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32853287

RESUMO

BACKGROUND: Vaccination is the most important preventive measure for protection against infectious diseases in humans and companion animals. Nevertheless, scepticism about the safety and importance of vaccines is increasing in human and in veterinary medicine. Although owner attitudes towards vaccination have been investigated in cats, there are no similar studies in dogs. The goals of this study were therefore to investigate the vaccination status of dogs in Germany, to determine owner compliance with vaccination and to identify factors that play a role in owners' decisions to have their dogs vaccinated. METHODS: Data were collected from August 2018 to February 2019 using an online survey targeting dog owners in Germany. A total of 3,881 questionnaires were evaluated, and factors associated with the vaccination status of dogs were determined by a linear logistic regression model using Akaike information criterion. Cohen's kappa statistic was used to evaluate agreement between questionnaire and 340 vaccination passports submitted voluntarily by owners. RESULTS: A total of 46.8% (n = 1,818/3,881) of dogs were vaccinated with core vaccines according to current guidelines with the lowest vaccination rate for leptospirosis (50.1%; n = 1,941/3,874). Dog's age (16 weeks to 15 months) (odds ratio (OR): 3.08; 95% CI: 2.05-4.68), type (working dog) (OR: 2.06; 95% CI: 1.22-3.53) and travelling abroad within previous 36 months (OR: 1.82; 95% CI: 1.12-2.96) had the strongest 'positive' association with the vaccination status. Recommendation from a veterinarian not to vaccinate against leptospirosis had the strongest 'negative' association (OR: 0.08; 95% CI: 0.04-0.18). CONCLUSION: The study revealed a need for improvement in vaccination compliance because of inadequate vaccination coverage, especially for leptospirosis, in dogs. Factors influencing owner compliance were numerous. Vaccination recommendations made by the veterinarian had a strong association with the vaccination status and should be used to increase canine vaccination rates.


Assuntos
Doenças do Cão/imunologia , Doenças do Cão/prevenção & controle , Propriedade/estatística & dados numéricos , Vacinação/estatística & dados numéricos , Animais , Atitude , Cães , Alemanha , Humanos , Leptospirose/imunologia , Leptospirose/prevenção & controle , Inquéritos e Questionários , Cobertura Vacinal/estatística & dados numéricos , Médicos Veterinários/estatística & dados numéricos
3.
Immunogenetics ; 72(5): 315-323, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32556497

RESUMO

Canine atopic dermatitis (CAD) is an inflammatory and pruritic allergic skin disease with both genetic and environmental risk factors described. We performed mRNA sequencing of non-lesional axillary skin biopsies from nine German shepherd dogs. Obtained RNA sequences were mapped to the dog genome (CanFam3.1) and a high-quality skin transcriptome was generated with 23,510 expressed gene transcripts. Differentially expressed genes (DEGs) were defined by comparing three controls to five treated CAD cases. Using a leave-one-out analysis, we identified seven DEGs: five known to encode proteins with functions related to an activated immune system (CD209, CLEC4G, LOC102156842 (lipopolysaccharide-binding protein-like), LOC480601 (regakine-1-like), LOC479668 (haptoglobin-like)), one (OBP) encoding an odorant-binding protein potentially connected to rhinitis, and the last (LOC607095) encoding a novel long non-coding RNA. Furthermore, high mRNA expression of inflammatory genes was found in axillary skin from an untreated mild CAD case compared with healthy skin. In conclusion, we define genes with different expression patterns in CAD case skin helping us understand post-treatment atopic skin. Further studies in larger sample sets are warranted to confirm and to transfer these results into clinical practice.


Assuntos
Dermatite Atópica/veterinária , Doenças do Cão/genética , Regulação da Expressão Gênica/imunologia , Pele/imunologia , Animais , Dermatite Atópica/genética , Dermatite Atópica/imunologia , Doenças do Cão/imunologia , Cães , Inflamação/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transcriptoma
5.
PLoS One ; 15(5): e0233305, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32413075

RESUMO

Despite vulnerability and unsanitary conditions of animal hoarding may predispose environmental contamination and spread of vectors and pathogens, no study to date has focused on their impact on public health and zoonotic diseases. Accordingly, this study aimed to assess the seroprevalence of anti-Toxoplasma gondii antibodies and associated factors in individuals with animal hoarding disorder (AHD) and their dogs in Curitiba, Southern Brazil. Blood samples were obtained from 264 dogs (21 households) and 19 individuals with AHD (11 households). Their blood was tested by indirect fluorescent antibody test (IFAT). Overall, anti-Toxoplasma gondii seropositivity was found in 21/264 dogs (7.95%; 95% CI: 4.69-11.22) with titers ranging from 16 to 4096, and in 7/19 individuals with AHD (36.84%; CI: 15.15-58.53) with titers ranging from 16 to 64. Serological analysis for anti-T. gondii antibodies were considered positive in at least one individual or dog in 9/11 (81.82%; 95% CI: 59.03-100.00) cases that were thoroughly assessed. Surprisingly, the seropositivity of individuals with AHD and their dogs was among the lowest reportedly observed in human and dog populations of Brazil. There was no significant association between positive owners and positive dogs or the presence of cats in the household. Regard epidemiological variables, a significant association was found between dog's seropositivity and the type of dog food. To the authors' knowledge, the present study represents the first investigation of T. gondii seroprevalence in individuals with hoarding disorder and their dogs. In conclusion, despite low sanitary conditions, anti-Toxoplasma gondii antibodies frequency in individuals with AHD and their dogs are lower than the general population likely due to low protozoan load in such isolated households.


Assuntos
Anticorpos Antiprotozoários/sangue , Doenças do Cão/epidemiologia , Transtorno de Acumulação/epidemiologia , Toxoplasma/imunologia , Toxoplasmose/epidemiologia , Ração Animal , Animais , Brasil/epidemiologia , Doenças do Cão/sangue , Doenças do Cão/imunologia , Cães , Geografia Médica , Colecionismo/sangue , Colecionismo/epidemiologia , Colecionismo/imunologia , Transtorno de Acumulação/sangue , Transtorno de Acumulação/imunologia , Humanos , Estudos Soroepidemiológicos , Análise Espacial , Toxoplasmose/sangue , Toxoplasmose/imunologia
6.
PLoS One ; 15(3): e0230272, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32168360

RESUMO

Canine prostate adenocarcinoma (PAC) and transitional cell carcinoma (TCC) of prostate and urinary bladder are highly invasive and metastatic tumors of closely neighbored organs. Cell lines are valuable tools to investigate tumor mechanisms and therapeutic approaches in vitro. PAC in dogs is infrequent, difficult to differentiate from TCC and usually characterized by poor prognosis, enhancing the value of the few available cell lines. However, as cell lines adapt to culturing conditions, a thorough characterization, ideally compared to original tissue, is indispensable. Herein, six canine PAC cell lines and three TCC cell lines were profiled by immunophenotype in comparison to respective original tumor tissues. Three of the six PAC cell lines were derived from primary tumor and metastases of the same patient. Further, two of the three TCC cell lines were derived from TCCs invading into or originating from the prostate. Cell biologic parameters as doubling times and chemoresistances to commonly used drugs in cancer treatment (doxorubicin, carboplatin and meloxicam) were assessed. All cell lines were immunohistochemically close to the respective original tissue. Compared to primary tumor cell lines, metastasis-derived cell lines were more chemoresistant to doxorubicin, but equally susceptive to carboplatin treatment. Two cell lines were multiresistant. COX-2 enzyme activity was demonstrated in all cell lines. However, meloxicam inhibited prostaglandin E2 production in only seven of nine cell lines and did neither influence metabolic activity, nor proliferation. The characterized nine cell lines represent excellent tools to investigate PAC as well as TCC in prostate and urinary bladder of the dog. Furthermore, the profiled paired cell lines from PAC primary tumor and metastasis provide the unique opportunity to investigate metastasis-associated changes PAC cells undergo in tumor progression. The combination of nine differently chemoresistant PAC and TCC cell lines resembles the heterogeneity of canine lower urinary tract cancer.


Assuntos
Adenocarcinoma/patologia , Carcinoma de Células de Transição/patologia , Neoplasias da Próstata/patologia , Neoplasias da Bexiga Urinária/patologia , Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/genética , Adenocarcinoma/imunologia , Animais , Carboplatina/farmacologia , Carcinoma de Células de Transição/tratamento farmacológico , Carcinoma de Células de Transição/genética , Carcinoma de Células de Transição/imunologia , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/imunologia , Proliferação de Células/efeitos dos fármacos , Ciclo-Oxigenase 2/genética , Doenças do Cão/tratamento farmacológico , Doenças do Cão/genética , Doenças do Cão/imunologia , Doenças do Cão/patologia , Cães , Doxorrubicina/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Imunofenotipagem , Masculino , Meloxicam/farmacologia , Metástase Neoplásica , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/genética , Neoplasias da Próstata/imunologia , Neoplasias da Bexiga Urinária/tratamento farmacológico , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/imunologia
7.
Parasite Immunol ; 42(6): e12713, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32173875

RESUMO

Canine leishmaniasis (CanL) is caused by the intracellular parasite Leishmania infantum. Prostaglandin E2 (PGE2 ) exerts potent regulatory effects on the immune system in experimental model Leishmania infection, but this influence has not yet been studied in CanL. In this study, PGE2 and PGE2 receptor levels and the regulatory effect of PGE2 on arginase activity, NO2 , IL-10, IL-17, IFN-γ, TNF-α and parasite load were evaluated in cultures of splenic leucocytes obtained from dogs with CanL in the presence of agonists and inhibitors. Our results showed that splenic leucocytes from dogs with CanL had lower EP2 receptor levels than those of splenic leucocytes from healthy animals. We observed that NO2 levels decreased when the cells were treated with a PGE2 receptor agonist (EP1/EP2/EP3) or COX-2 inhibitor (NS-398) and that TNF-α, IL-17 and IFN-γ cytokine levels decreased when the cells were treated with a PGE2 receptor agonist (EP2) or PGE2 itself. The parasite load in splenic leucocyte cell cultures from dogs with CanL decreased after stimulation of the cells with PGE2 . We conclude that Leishmania infection of dogs modulates PGE2 receptors and speculate that the binding of PGE2 to its receptors may activate the microbicidal capacity of cells.


Assuntos
Citocinas/imunologia , Dinoprostona/metabolismo , Doenças do Cão/tratamento farmacológico , Leishmania infantum/imunologia , Leishmaniose/veterinária , Receptores de Prostaglandina E/metabolismo , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Inibidores de Ciclo-Oxigenase 2/farmacologia , Dinoprostona/agonistas , Dinoprostona/antagonistas & inibidores , Doenças do Cão/imunologia , Doenças do Cão/parasitologia , Cães , Leishmaniose/tratamento farmacológico , Leishmaniose/imunologia , Óxido Nítrico/análise , Nitrobenzenos/farmacologia , Carga Parasitária , Receptores de Prostaglandina E/agonistas , Receptores de Prostaglandina E/fisiologia , Sulfonamidas/farmacologia , Fator de Necrose Tumoral alfa/imunologia
8.
J Vet Med Sci ; 82(4): 483-487, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32147621

RESUMO

DEP domain-containing 1B (DEPDC1B) is involved in the regulation of cell de-adhesion and actin cytoskeleton activity during the G2/M transition of the cell cycle, and its overexpression has been proven to be associated with cancer progression in several human cancers. Canine DEPDC1B was identified as a gene that was overexpressed in canine lymphoma tissues in our previous study. However, in dogs, the protein expression of DEPDC1B remains to be determined due to the lack of a specific monoclonal antibody. Here, we developed rat monoclonal antibodies against canine DEPDC1B and characterized their applicability for immunodetection assays. Our findings demonstrated that these antibodies are functional and can be important tools to investigate the precise role of DEPDC1B in canine tumors.


Assuntos
Anticorpos Monoclonais/imunologia , Proteínas Ativadoras de GTPase/imunologia , Animais , Western Blotting/veterinária , Linhagem Celular Tumoral , Doenças do Cão/imunologia , Cães/imunologia , Feminino , Proteínas Ativadoras de GTPase/genética , Imuno-Histoquímica/métodos , Imuno-Histoquímica/veterinária , Imunoprecipitação/métodos , Imunoprecipitação/veterinária , Linfoma de Células T/veterinária , Células Madin Darby de Rim Canino , Camundongos , Ratos Sprague-Dawley , Proteínas Recombinantes/imunologia
9.
Parasite Immunol ; 42(5): e12706, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32119124

RESUMO

To provide useful information based on the macropathology, histopathology and immunohistochemical investigation in the spleens of dogs with Babesia rossi infection. Control spleens were collected from four healthy dogs euthanized for welfare reasons. Nine dogs that died naturally because of a mono-infection with Babesia rossi were selected for the diseased group. One haematoxylin-and-eosin-stained section of splenic tissue from each of the infected and control dogs was examined under the light microscope. Immunohistochemical markers were applied to characterize different immunocyte populations. The application of analytic software enabled semi-quantitative comparison of leucocyte subpopulations. Routine splenic histopathology revealed diffuse intermingling of white and red pulp from infected dogs with a clear loss of distinction between these zones. Immunohistochemistry revealed an increase in the proportion of tissue resident and bone marrow origin macrophages in the infected spleens. Apart from a few remnant lymphocytes within the peri-arteriolar lymphatic sheaths and follicles, the majority of the immunocytes redistributed to the red pulp, supporting the observation of white and red pulp intermingling. The majority of our findings are in agreement with histomorphological descriptions of the spleen in a variety of noncanid mammalian hosts with lethal malaria or babesiosis.


Assuntos
Babesia/fisiologia , Babesiose/patologia , Doenças do Cão/patologia , Baço/patologia , Animais , Babesiose/imunologia , Babesiose/parasitologia , Doenças do Cão/imunologia , Doenças do Cão/parasitologia , Cães , Leucócitos/imunologia , Leucócitos/parasitologia , Linfócitos/imunologia , Linfócitos/parasitologia , Baço/imunologia , Baço/parasitologia
10.
Vet Immunol Immunopathol ; 221: 110015, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32058160

RESUMO

Atopic dermatitis (AD) is an allergic skin disease that causes significant morbidity and affects multiple species. AD is highly prevalent in companion dogs, and the clinical management of the disease remains challenging. An improved understanding of the immunologic and genetic pathways that lead to disease could inform the development of novel treatments. In allergic humans and mouse models of AD, the disease is associated with Th2 and group 2 innate lymphoid cell (ILC2) activation that drives type 2 inflammation. Type 2 inflammation also appears to be associated with AD in dogs, but gaps remain in our understanding of how key type 2-associated cell types such as canine Th2 cells and ILC2s contribute to the pathogenesis of canine AD. Here, we describe previously uncharacterized canine ILC2-like cells and Th2 cells ex vivo that produced type 2 cytokines and expressed the transcription factor Gata3. Increased circulating Th2 cells were associated with chronic canine AD. Single-cell RNA sequencing revealed a unique gene expression signature in T cells in dogs with AD. These findings underline the importance of pro-allergic Th2 cells in orchestrating AD and provide new methods and pathways that can inform the development of improved therapies.


Assuntos
Dermatite Atópica/veterinária , Doenças do Cão/imunologia , Imunidade Inata , Linfócitos/imunologia , Células Th2/imunologia , Animais , Células Sanguíneas/imunologia , Dermatite Atópica/imunologia , Cães , Feminino , Inflamação , Linfócitos/classificação , Masculino , Análise de Sequência de RNA , Análise de Célula Única
11.
Int J Parasitol ; 50(2): 103-110, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32004510

RESUMO

Babesia rossi is an important, tick-borne intraerythrocytic protozoan parasite; however, its natural history and epidemiology is poorly understood. Babesia rossi is the most virulent Babesia sp. in domestic dogs and is generally considered to cause severe babesiosis, which is fatal if left untreated. However, subclinical infections and mild disease from B. rossi have been reported, although the clinical progression of these cases was not reported. Therefore, to better understand B. rossi under field conditions, we evaluated its clinical progression and seroprevalence in an owned, free-roaming dog population in Zenzele, South Africa, where the parasite is endemic and prevention is not routine. The entire dog population in Zenzele was monitored intensively at the individual level from March 2008 until April 2014, primarily for a longitudinal study on rabies control. Subsequent evaluation of B. rossi comprised analyses of clinical and laboratory data collected from the Zenzele dog population during the 6 year study period. A substantial proportion (31% (n = 34)) of 109 dogs (randomly selected from every available dog in February/March 2010 older than ~6-8 weeks (n = 246)) tested by Indirect Fluorescent Antibody Test had seroconverted strongly to B. rossi. All 34 dogs were generally consistently healthy adults, determined from regular clinical examinations between March 2008 and April 2014. Blood smear examinations at multiple time points between July 2009 and February 2011 were also undertaken for almost all of these (34) seropositive dogs and all those tested were consistently negative for Babesia spp. Subclinical infections and mild disease were also the main findings for a separate group of 18 dogs positive for Babesia spp. on blood smear examination and confirmed to be infected with B. rossi by Polymerase Chain Reaction - Reverse Line Blot. Almost all of these dogs were positive at only one time point from repeat blood smear examinations between July 2009 and February 2011. We suggest that these observations are consistent with immunity acquired from repeated, low-level exposure to the parasite, generating transient subclinical infections or mild disease. Should this be the case, the use of tick control, particularly in adult dogs in free-roaming populations in B. rossi endemic regions, should be carefully considered.


Assuntos
Babesia/isolamento & purificação , Babesiose , Doenças do Cão/parasitologia , Estudos Soroepidemiológicos , Imunidade Adaptativa , Animais , Vetores Artrópodes/parasitologia , Babesiose/imunologia , Babesiose/parasitologia , Babesiose/transmissão , Doenças do Cão/imunologia , Doenças do Cão/transmissão , Cães , Seguimentos , Estudos Longitudinais , Patologia Molecular , Reação em Cadeia da Polimerase/veterinária , Testes Sorológicos , África do Sul/epidemiologia , Controle de Ácaros e Carrapatos , Carrapatos/parasitologia
12.
Parasit Vectors ; 13(1): 69, 2020 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-32054524

RESUMO

BACKGROUND: Canine heartworm disease, caused by Dirofilaria immitis, has global veterinary importance. In Australia, the prevalence of canine heartworm infection decreased markedly following the introduction of over-the-counter macrocyclic lactones. We aimed to estimate the prevalence of canine heartworm infection in at-risk populations of dogs in eastern Australia and analyse published prevalence data from Australia. METHODS: In total, 566 dogs from eastern Australia were tested for the presence of D. immitis antigen. Four cohorts were studied: pig-hunting dogs from Queensland (Cohort 1, n = 104), dogs from remote New South Wales (NSW) (Cohort 2, n = 332), urban pets from rural NSW (Cohort 3, n = 45) and ex-racing Greyhounds from Sydney, NSW (Cohort 4, n = 85). Serum samples were screened for D. immitis antigen using a reference laboratory microwell-based assay (DiroChek®) or a point-of-care immunochromatography test kit (Anigen Rapid®). Risk factors associated with the odds of D. immitis antigen seropositivity were identified using binary logistic regression models. Seropositive blood samples were tested for the presence and quantity of D. immitis DNA using a species specific real-time (q)PCR assay. A metanalysis of the Australian canine heartworm literature was conducted. RESULTS: The prevalence of dirofilariasis in pig-hunting dogs from Queensland (Cohort 1) was 12.5% (95% CI: 6.5-18.9%), with a subpopulation of dogs from Central Queensland having a prevalence of 21% (95% CI: 12.3-33.4%). Age was significantly associated with D. immitis antigen seropositivity (increased risk with increased age). The odds of being > 5 years versus ≤ 5 years was 3.7-times (95% CI: 1.1-12.5) greater in antigen positive versus antigen negative dogs. No D. immitis antigen positive dogs were detected in dogs from NSW (Cohorts 2-4). The Australian canine heartworm disease literature includes 98 peer-reviewed publications (1901-2019) with 30 studies reporting on D. immitis prevalence in dogs. Throughout the publication peak period (1980s), the primary antemortem diagnostic test was detection of microfilariae. CONCLUSIONS: Canine heartworm infection in dogs used for pig hunting is a previously unexplored topic in Australia. Pig-hunting dogs are infected with canine heartworm in Queensland, Australia, placing pet dogs and cats at increased risk of infection.


Assuntos
Dirofilariose/epidemiologia , Doenças do Cão/parasitologia , Cães/parasitologia , Fatores Etários , Animais , Antígenos de Helmintos/imunologia , Estudos de Coortes , Dirofilaria immitis/imunologia , Dirofilaria immitis/isolamento & purificação , Dirofilariose/imunologia , Doenças do Cão/epidemiologia , Doenças do Cão/imunologia , Feminino , Masculino , Comportamento Predatório , Prevalência , Queensland/epidemiologia , Suínos
13.
Res Vet Sci ; 130: 19-25, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32109759

RESUMO

Immune control of Leishmania infantum, the causative agent of most canine leishmaniosis (CanL), requires a balancing act between inflammatory and regulatory responses. This balance is specifically between the proinflammatory T helper 1 type (Th1) CD4+ T cells that are responsible for controlling parasite replication and T regulatory 1 cells which mediate an immunosuppressive, regulatory, response needed to dampen overabundant inflammation but if predominant, result in CanL progression. How this delicate immune cell interaction occurs in the dog will be highlighted in this review, focusing on the progressive changes observed within myeloid lineage cells (predominantly macrophages), B cells and T cells. After exposure to parasites, macrophages should become activated, eliminating L. infantum through release of reactive oxygen species. Unfortunately, multiple parasite and host factors can prevent macrophage activation allowing parasites to persist within them. T cells balance between a productive TH1 type CD4+ response capable of producing IFN-γ which aids macrophage activation versus T cell exhaustion which reduces T cell proliferation, IFN-γ production and allows parasite expansion within macrophages. Neutrophils and Th17 cells add to the inflammatory state, aiding in parasite removal, but also leading to pathology. A regulatory B cell population increases IL-10 production and down regulates the TH1 response allowing parasite growth. All of these immune challenges affect the balance between progression to clinical disease and maintaining sub-clinical disease. Vaccines and immunotherapies targeted at recovering or maintaining T and B cell function can be important factors in mending the immune balance required to survive CanL.


Assuntos
Linfócitos B/imunologia , Doenças do Cão/imunologia , Leishmaniose Visceral/veterinária , Macrófagos/imunologia , Linfócitos T/imunologia , Animais , Cães , Leishmania infantum/fisiologia , Leishmaniose Visceral/imunologia
14.
PLoS Negl Trop Dis ; 14(1): e0008021, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31961868

RESUMO

Domestic dogs are the main reservoir of Leishmania infantum, a causative agent of visceral leishmaniasis (VL). The number of human disease cases is associated with the rate of canine infection. Currently available drugs are not efficient at treating canine leishmaniasis (CanL) and months after the treatment most dogs show disease relapse, therefore the development of new drugs or new therapeutic strategies should be sought. In CanL, dogs lack the ability to mount a specific cellular immune response suitable for combating the parasite and manipulation of cytokine signaling pathway has the potential to form part of effective immunotherapeutic methods. In this study, recombinant canine cytokines (rcaIL-12, rcaIL-2, rcaIL-15 and rcaIL-7) and soluble receptor IL-10R1 (rcasIL-10R1), with antagonistic activity, were evaluated for the first time in combination (rcaIL-12/rcaIL-2, rcaIL-12/rcaIL-15, rcaIL-12/rcasIL-10R1, rcaIL-15/rcaIL-7) or alone (rcasIL-10R1) to evaluate their immunomodulatory capacity in peripheral blood mononuclear cells (PBMCs) from dogs with leishmaniasis. All the combinations of recombinant proteins tested were shown to improve lymphoproliferative response. Further, the combinations rcaIL-12/rcaIL-2 and rcaIL-12/rcaIL-15 promoted a decrease in programmed cell death protein 1 (PD-1) expression in lymphocytes. These same combinations of cytokines and rcaIL-12/rcasIL-10R1 induced IFN-γ and TNF-α production in PBMCs. Furthermore, the combination IL-12/IL-15 led to an increased in T-bet expression in lymphocytes. These findings are encouraging and indicate the use of rcaIL-12 and rcaIL-15 in future in vivo studies aimed at achieving polarization of cellular immune responses in dogs with leishmaniasis, which may contribute to the development of an effective treatment against CanL.


Assuntos
Doenças do Cão/tratamento farmacológico , Doenças do Cão/imunologia , Interleucina-12/administração & dosagem , Interleucina-15/administração & dosagem , Leishmaniose Visceral/imunologia , Animais , Doenças do Cão/genética , Doenças do Cão/parasitologia , Cães , Imunidade Celular , Interferon gama/genética , Interferon gama/imunologia , Interleucina-12/genética , Interleucina-12/imunologia , Interleucina-15/genética , Interleucina-15/imunologia , Leishmania infantum/fisiologia , Leishmaniose Visceral/tratamento farmacológico , Leishmaniose Visceral/parasitologia , Leishmaniose Visceral/veterinária , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/parasitologia , Receptor de Morte Celular Programada 1/genética , Receptor de Morte Celular Programada 1/imunologia , Receptores de Citocinas/genética , Receptores de Citocinas/imunologia
15.
PLoS One ; 15(1): e0227386, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31923271

RESUMO

Canine Chronic Ulcerative Stomatitis is a spontaneously occurring inflammatory disease of the oral mucosa. An immune-mediated pathogenesis is suspected though not yet proven. We have recently reported on the clinical and histologic features, and identification of select leukocyte cell populations within the lesion. A clinical and histologic similarity to oral lichen planus of people was proposed. In the present study, these initial observations are extended by examining lesions from 24 dogs with clinical evidence of chronic ulcerative stomatitis. Because dogs with chronic ulcerative stomatitis often have concurrent periodontal disease, we wondered if dental plaque/biofilm may be a common instigator of inflammation in both lesions. We hypothesized that dogs with chronic ulcerative stomatitis would exhibit a spectrum of pathologic changes and phenotype of infiltrating leukocytes that would inform lesion pathogenesis and that these changes would differ from inflammatory phenotypes in periodontitis. Previously we identified chronic ulcerative stomatitis lesions to be rich in FoxP3+ and IL17+ cells. As such, we suspect that these leukocytes play an important role in lesion pathogenesis. The current study confirms the presence of moderate to large numbers of FoxP3+ T cells and IL17+ cells in all ulcerative stomatitis lesions using confocal immunofluorescence. Interestingly, the majority of IL17+ cells were determined to be non-T cells and IL17+ cell frequencies were negatively correlated with severity on the clinical scoring system. Three histologic subtypes of ulcerative stomatitis were determined; lichenoid, deep stomatitis and granulomatous. Periodontitis lesions, like stomatitis lesions, were B cell and plasma cell rich, but otherwise differed from the stomatitis lesions. Direct immunofluorescence results did not support an autoantibody-mediated autoimmune disease process. This investigation contributes to the body of literature regarding leukocyte involvement in canine idiopathic inflammatory disease pathogenesis.


Assuntos
Doenças do Cão/imunologia , Gengivite Ulcerativa Necrosante/imunologia , Animais , Doença Crônica , Diagnóstico Diferencial , Doenças do Cão/patologia , Cães , Técnica Direta de Fluorescência para Anticorpo , Gengivite Ulcerativa Necrosante/diagnóstico , Gengivite Ulcerativa Necrosante/patologia , Gengivite Ulcerativa Necrosante/veterinária , Inflamação/etiologia , Leucócitos/patologia , Mucosa Bucal/patologia , Doenças Periodontais/diagnóstico
16.
Vet Immunol Immunopathol ; 221: 110012, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31978678

RESUMO

Alterations in serum cytokine levels and profiles have been reported in association with a variety of disease conditions (e.g., allergic, immune-mediated, etc.) in both humans and animals. In comparison to serum cytokine measurements, tear cytokine measurements might be expected to more accurately reflect the inflammatory milieu associated with periocular disease. The purpose of this study was to use a multiplexed assay to compare the cytokine profile of tears in healthy dogs to those with inflammatory skin and periocular disease. We were able to detect IL-2, IL-6, IL-8, and TNF-α in >47 % of tear samples from both healthy canine patients and those with inflammatory dermatologic disease (with or without concurrent periocular involvement). In contrast, IL-7, IL-10 and IFN-γ were rarely detected. Dogs with both dermatologic and periocular disease (but not dermatologic disease alone) had higher levels of IL-8 (P < 0.001, P > 0.05, respectively) relative to healthy dogs. Patients with concurrent dermatologic and periocular disease also demonstrated significantly greater variability in IL-8 concentrations between eyes than did healthy dogs (P < 0.0001). Our findings suggest that tear cytokine analysis may prove to be a useful tool to investigate the role and interactions of the local ocular immune response in patients with inflammatory periocular disease.


Assuntos
Citocinas/imunologia , Doenças do Cão/imunologia , Oftalmopatias/veterinária , Dermatopatias/veterinária , Lágrimas/imunologia , Animais , Cães , Olho/imunologia , Oftalmopatias/imunologia , Feminino , Interleucina-2/imunologia , Interleucina-6/imunologia , Interleucina-8/imunologia , Masculino , Dermatopatias/imunologia , Fatores de Necrose Tumoral/imunologia
17.
Vet Immunol Immunopathol ; 221: 110010, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31981823

RESUMO

Macrophage migration inhibitory factor (MIF) is a pleiotropic cytokine that is produced by many cell types in situations of homeostasis or disease. One of its functions is to act as a proinflammatory molecule. In humans, several studies have shown that MIF levels become elevated in the serum, urine, cerebrospinal fluid and tissues of patients with chronic inflammatory diseases (systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, sepsis, atheromas, diabetes and cancer). In dogs, distemper is a viral infectious condition that may lead to demyelination and inflammation in the central nervous system (CNS). In addition to the action of the virus, the inflammatory process may give rise to lesions in the white matter. Therefore, the objectives of the present study were to evaluate the role of MIF in the encephalitis that the canine distemper virus causes and to compare this with immunodetection of major histocompatibility complex-II (MHC-II), CD3 T lymphocytes, MMP-9 and glial fibrillary acidic protein (GFAP; astrocytes) in demyelinated areas of the encephalon, in order to ascertain whether these findings might be related to the severity of the encephalic lesions. To this end, a retrospective study on archived paraffinized blocks was conducted, in which 21 encephala from dogs that had been naturally infected with the canine distemper virus (infected group) and five from dogs that had been free from systemic or CNS-affecting diseases (control group) were used. In the immunohistochemical analysis on the samples, the degree of marking by GFAP, MHC-II, MMP-9 and MIF was greater in the demyelinated areas and in the adjacent neuropil, and this was seen particularly in astrocytes. Detection of CD3 was limited to perivascular cuffs. In areas of liquefactive necrosis, Gitter cells were positive for MMP-9, MIF and MHC-II. Hence, it was concluded that activated astrocytes influenced the afflux of T lymphocytes to the encephalon (encephalitis). In the more advanced phases, activated phagocytes in the areas of liquefactive necrosis (Gitter cells) continued to produce inflammatory mediators even after the astrocytes in these localities had died, thereby worsening the encephalic lesions. Distemper virus-activated astrocytes and microglia produce MIF that results in proinflammatory stimulus on glial cells and brain-infiltrating leukocytes. Therefore, the effect of the inflammatory response is potentiated on the neuropil, resulting in neurological clinical signs.


Assuntos
Astrócitos/química , Cinomose/imunologia , Doenças do Cão/imunologia , Doenças do Cão/virologia , Encefalite/veterinária , Fatores Inibidores da Migração de Macrófagos/imunologia , Animais , Astrócitos/imunologia , Vírus da Cinomose Canina , Cães , Encefalite/imunologia , Encefalite/virologia , Técnicas Histológicas , Inclusão em Parafina , Estudos Retrospectivos
18.
J Vasc Interv Radiol ; 31(3): 482-491.e4, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31956003

RESUMO

PURPOSE: To determine the safety and feasibility of percutaneous high-frequency irreversible electroporation (HFIRE) for primary liver cancer and evaluate the HFIRE-induced local immune response. MATERIALS AND METHODS: HFIRE therapy was delivered percutaneously in 3 canine patients with resectable hepatocellular carcinoma (HCC) in the absence of intraoperative paralytic agents or cardiac synchronization. Pre- and post-HFIRE biopsy samples were processed with histopathology and immunohistochemistry for CD3, CD4, CD8, and CD79a. Blood was collected on days 0, 2, and 4 for complete blood count and chemistry. Numeric models were developed to determine the treatment-specific lethal thresholds for malignant canine liver tissue and healthy porcine liver tissue. RESULTS: HFIRE resulted in predictable ablation volumes as assessed by posttreatment CT. No detectable cardiac interference and minimal muscle contraction occurred during HFIRE. No clinically significant adverse events occurred secondary to HFIRE. Microscopically, a well-defined ablation zone surrounded by a reactive zone was evident in the majority of samples. This zone was composed primarily of maturing collagen interspersed with CD3+/CD4-/CD8- lymphocytes in a proinflammatory microenvironment. The average ablation volumes for the canine HCC patients and the healthy porcine tissue were 3.89 cm3 ± 0.74 and 1.56 cm3 ± 0.16, respectively (P = .03), and the respective average lethal thresholds were 710 V/cm ± 28.2 and 957 V/cm ± 24.4 V/cm (P = .0004). CONCLUSIONS: HFIRE can safely and effectively be delivered percutaneously, results in a predictable ablation volume, and is associated with lymphocytic tumor infiltration. This is the first step toward the use of HFIRE for treatment of unresectable liver tumors.


Assuntos
Técnicas de Ablação/veterinária , Carcinoma Hepatocelular/veterinária , Doenças do Cão/cirurgia , Eletroporação/veterinária , Neoplasias Hepáticas/veterinária , Animais , Complexo CD3/imunologia , Carcinoma Hepatocelular/imunologia , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/cirurgia , Doenças do Cão/imunologia , Doenças do Cão/patologia , Cães , Estudos de Viabilidade , Feminino , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , Neoplasias Hepáticas/imunologia , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/cirurgia , Linfócitos do Interstício Tumoral/imunologia , Masculino , Estudo de Prova de Conceito , Sus scrofa
19.
Vet Dermatol ; 31(1): 5-27, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31957202

RESUMO

BACKGROUND: Demodicosis is a common disease in small animal veterinary practice worldwide with a variety of diagnostic and therapeutic options. OBJECTIVES: To provide consensus recommendations on the diagnosis, prevention and treatment of demodicosis in dogs and cats. METHODS AND MATERIALS: The authors served as a Guideline Panel (GP) and reviewed the literature available before December 2018. The GP prepared a detailed literature review and made recommendations on selected topics. A draft of the document was presented at the North American Veterinary Dermatology Forum in Maui, HI, USA (May 2018) and at the European Veterinary Dermatology Congress in Dubrovnik, Croatia (September 2018) and was made available via the World Wide Web to the member organizations of the World Association for Veterinary Dermatology for a period of three months. Comments were solicited and responses were incorporated into the final document. CONCLUSIONS: In young dogs with generalized demodicosis, genetic and immunological factors seem to play a role in the pathogenesis and affected dogs should not be bred. In old dogs and cats, underlying immunosuppressive conditions contributing to demodicosis should be explored. Deep skin scrapings are the diagnostic gold standard for demodicosis, but trichograms and tape squeeze preparations may also be useful under certain circumstances. Amitraz, macrocyclic lactones and more recently isoxazolines have all demonstrated good efficacy in the treatment of canine demodicosis. Therapeutic selection should be guided by local drug legislation, drug availability and individual case parameters. Evidence for successful treatment of feline demodicosis is strongest for lime sulfur dips and amitraz baths.


Assuntos
Doenças do Gato/diagnóstico , Doenças do Gato/tratamento farmacológico , Dermatite/veterinária , Doenças do Cão/diagnóstico , Doenças do Cão/tratamento farmacológico , Infestações por Ácaros/veterinária , Animais , Doenças do Gato/imunologia , Gatos , Dermatite/imunologia , Dermatite/parasitologia , Doenças do Cão/imunologia , Cães , Inseticidas/uso terapêutico , Infestações por Ácaros/diagnóstico , Infestações por Ácaros/tratamento farmacológico , Infestações por Ácaros/imunologia , Ácaros/efeitos dos fármacos , Pele/efeitos dos fármacos , Pele/parasitologia , Pele/patologia , Medicina Veterinária/métodos , Medicina Veterinária/organização & administração
20.
J Vet Med Sci ; 82(2): 177-183, 2020 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-31875597

RESUMO

Hydrolyzed proteins are often prescribed for dogs with food hypersensitivity in food elimination programs. However, the potential of these diets to stimulate lymphocyte-mediated hypersensitivity is currently unknown. In this study, two commercially available hydrolyzed diets for dogs, D-1 (Aminopeptide Formula Dry, Royal Canin Japon, Tokyo, Japan), and D-2 (Canine z/d Ultra Dry, Hill's-Colgate (Japan) Ltd., Tokyo, Japan), were analyzed to identify residual proteins or peptides, as well as activated helper T-lymphocyte reactions in dogs with suspected food hypersensitivity. Proteins and peptides with molecular weights >1 kDa (majority 1.5-3.5 kDa) were detected in both diet extracts with sodium dodecyl sulfate polyacrylamide gel electrophoresis, and size exclusion chromatography. When peripheral blood mononuclear cells (PBMC's) from 316 dogs with suspected food allergies were cultured with hydrolyzed diet extracts, flow cytometry analysis revealed detectable levels of CD25low helper T-lymphocytes stimulated by D-1 and D-2 in 91 of 316, (28.8%), and 75 of 316 (23.7%) samples, respectively. These data indicated that the extracts contained proteins or peptides large enough to activate the lymphocytes. The percentages of CD25low helper T-lymphocytes stimulated by D-1 and D-2 extracts increased to 38.7% and 29.6%, respectively, in 186 of the original 316 samples (186/316, 58.9%), also reactive to poultry-related antigens. Thus, both poultry-related antigens, and D-1 and D-2 diet extracts may activate helper T-lymphocytes. These results demonstrate that hydrolyzed diets may contain proteins that stimulate helper T-lymphocytes, and may not be effective for treating all dogs with food hypersensitivity.


Assuntos
Ração Animal/análise , Doenças do Cão/imunologia , Hipersensibilidade Alimentar/veterinária , Linfócitos T Auxiliares-Indutores/imunologia , Animais , Dieta/veterinária , Proteínas na Dieta/imunologia , Doenças do Cão/sangue , Cães , Hipersensibilidade Alimentar/sangue , Hipersensibilidade Alimentar/imunologia , Ativação Linfocitária/imunologia , Hidrolisados de Proteína
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