RESUMEN
BACKGROUND: Canine atopic dermatitis (cAD) is a disease associated with Type 2 helper T (Th2) immune responses in the acute phase of the disease. In humans, keratinocytes are activated by Th2 cytokines via the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway. However, the activation of keratinocytes by Th2 cytokines in cAD has not yet been demonstrated. HYPOTHESIS/OBJECTIVES: To evaluate keratinocyte activation based on the phosphorylation (p) of JAK1, STAT3 and STAT6. ANIMALS: Seven dogs with cAD and three healthy dogs. MATERIALS AND METHODS: Immunohistochemical analysis was performed to detect pJAK1, pSTAT3 and pSTAT6 in keratinocytes in normal canine skin, and the skin of atopic dogs. In the latter group samples were collected from both primary and secondary lesions, and nonaffected skin. RESULTS: The percentage of pJAK1-positive keratinocytes was significantly higher in primary cAD lesions than in healthy skin (p < 0.05). No significant differences were observed in pSTAT3-positive keratinocytes among the groups. The percentage of pSTAT6-positive keratinocytes was significantly higher in primary and secondary lesions than in healthy skin (p < 0.05, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: The novel finding in this study was the activation of keratinocytes as demonstrated by the phosphorylation of JAK1/STATs in lesional and nonlesional cAD skin. These results suggest the potential of not only JAK1, but also of STAT6 as therapeutic targets for cAD.
Asunto(s)
Dermatitis Atópica , Enfermedades de los Perros , Humanos , Perros , Animales , Dermatitis Atópica/tratamiento farmacológico , Dermatitis Atópica/veterinaria , Janus Quinasa 1/metabolismo , Fosforilación , Factor de Transcripción STAT3/metabolismo , Factor de Transcripción STAT3/uso terapéutico , Queratinocitos , Citocinas/metabolismo , Enfermedades de los Perros/patologíaRESUMEN
BACKGROUND: In human medicine, narrow-band ultraviolet B (NB-UVB) phototherapy has been used to treat various T-cell-mediated skin diseases. However, the effect of NB-UVB on inflamed canine skin remains uncertain. OBJECTIVES: To investigate the effect of NB-UVB phototherapy on the skin of dogs with hapten-induced contact dermatitis. ANIMALS: Seven healthy beagles without skin problems. METHODS AND MATERIALS: Dogs were irradiated with varying doses of NB-UVB to determine the minimal erythema dose (MED). After determining the MEDs of six dogs (excluding one of the seven whose skin did not show a visible reaction), we investigated the effect of NB-UVB on their inflamed skin by topically applying 2,4-dinitrochlorobenzene (DNCB), which causes type 1 helper T cell (Th1)- and cytotoxic T-cell (Tc)1-induced skin inflammation. We then irradiated the skin with NB-UVB. We analysed the treated skin samples via histopathological and immunohistochemical methods, and TdT-mediated dUTP nick-end labelling (TUNEL) to demonstrate apoptotic cells. We also analysed the cytokine gene transcription via real-time quantitative reverse transcription PCR. RESULTS: The NB-UVB MEDs caused mild inflammatory changes yet no severe epidermal exfoliations in the irradiated skin. In DNCB-treated skin irradiated by the NB-UVB MEDs, TUNEL-positive dermal apoptotic cells were increased significantly compared with those of DNCB-treated, nonirradiated skin. INF-γ and TNF-α transcription levels in DNCB-treated, irradiated skin were significantly lower than those in the DNCB-treated, nonirradiated skin. CONCLUSION AND CLINICAL RELEVANCE: Phototherapy using NB-UVB MEDs attenuated cutaneous Th1 and Tc1 cytokine responses with minimal skin damage in a canine model of hapten-induced contact dermatitis.
Asunto(s)
Dermatitis por Contacto , Enfermedades de los Perros , Terapia Ultravioleta , Animales , Dermatitis por Contacto/veterinaria , Enfermedades de los Perros/radioterapia , Perros , Haptenos , Piel , Linfocitos T , Rayos Ultravioleta/efectos adversos , Terapia Ultravioleta/efectos adversos , Terapia Ultravioleta/veterinariaRESUMEN
BACKGROUND: Thymus and activation-regulated chemokine (TARC/CCL17) has been implicated in the pathogenesis of canine atopic dermatitis (cAD). Serum TARC concentrations are a reliable biomarker for human atopic dermatitis; however, their potential as a biomarker for cAD has not been investigated. HYPOTHESIS/OBJECTIVES: To investigate whether serum TARC concentrations correlate with disease severity and therapeutic responses for cAD. ANIMALS: Thirty-nine dogs with cAD and 42 healthy dogs were recruited. METHODS AND MATERIALS: Serum TARC concentrations in dogs with cAD and healthy dogs were measured by sandwich ELISA with anti-canine TARC antibodies. The clinical severity of cAD was scored using the validated Canine Atopic Dermatitis Extent and Severity Index, 4th iteration (CADESI-04). Serum TARC concentrations were compared between dogs with cAD and healthy controls, and their relationship with CADESI-04 was examined. Serum TARC concentrations also were measured in 20 dogs with cAD treated with prednisolone or oclacitinib for four weeks. RESULTS: Serum TARC concentrations were significantly higher in dogs with cAD than in healthy dogs (P < 0.001). In dogs with cAD, serum TARC concentrations correlated with CADESI-04 scores (ρ = 0.457, P < 0.01). Furthermore, serum TARC concentrations significantly decreased in treated dogs with the attenuation of clinical signs (P < 0.001). Changes in serum TARC concentrations before and after treatment correlated with those in CADESI-04 scores (ρ = 0.746, P < 0.001). CONCLUSIONS AND CLINICAL RELEVANCE: Serum TARC concentrations have potential as a clinical and research tool for the objective evaluation of disease severity and therapeutic responses for cAD.
Asunto(s)
Dermatitis Atópica , Enfermedades de los Perros , Animales , Biomarcadores , Quimiocina CCL17 , Dermatitis Atópica/diagnóstico , Dermatitis Atópica/veterinaria , Enfermedades de los Perros/diagnóstico , Perros , Femenino , Masculino , Prednisolona , Índice de Severidad de la EnfermedadRESUMEN
BACKGROUND: Degenerative myelopathy (DM) is a progressive neurodegenerative disease frequently found in Pembroke Welsh Corgis (PWCs). Most DM-affected PWCs are homozygous for the mutant superoxide dismutase 1 (SOD1) allele; however, the genetic examination for the SOD1 mutation does not exclusively detect symptomatic dogs. In order to identify novel biomarkers, the plasma microRNA (miRNA) profiles of PWCs with DM were investigated. RESULTS: Quantification of the plasma levels of 277 miRNAs by an RT-qPCR array identified 11 up-regulated miRNAs and 7 down-regulated miRNAs in DM-affected PWCs from those in wild-type SOD1 PWCs. A pathway analysis identified 3 miRNAs: miR-26b, miR-181a, and miR-196a, which potentially regulate several genes associated with SOD1. In order to validate the diagnostic accuracy of the candidate miRNAs in the aged PWC population, candidate miRNAs in plasma were measured by RT-qPCR and a receiver operating characteristic (ROC) curve analysis was performed. miR-26b had the largest area under the ROC curve for distinguishing DM PWCs from healthy PWCs (sensitivity, 66.7%; specificity, 87.0%). The plasma level of miR-26b was significantly higher in the DM group than in the healthy control group. A positive correlation was observed between increases in the plasma level of miR-26b and disease progression. CONCLUSIONS: These results suggest that plasma miR-26b is a potential novel diagnostic biomarker of DM.
Asunto(s)
Enfermedades de los Perros/genética , MicroARNs/sangre , Enfermedades Neurodegenerativas/veterinaria , Animales , Biomarcadores/sangre , Progresión de la Enfermedad , Enfermedades de los Perros/diagnóstico , Perros , Femenino , Masculino , Mutación , Enfermedades Neurodegenerativas/sangre , Enfermedades Neurodegenerativas/diagnóstico , Enfermedades Neurodegenerativas/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Superóxido Dismutasa-1/genéticaRESUMEN
OBJECTIVE: To evaluate the accuracy and safety of a novel patient-specific drill guide template for stabilizing the thoracolumbar vertebrae of dogs. STUDY DESIGN: Cadaveric experimental study and prospective case series. SAMPLE POPULATION: Cadaveric canine thoracolumbar vertebral specimens (n = 3) and clinical cases of thoracolumbar spinal instability (n = 4). METHODS: Computed tomography data of the thoracolumbar spines were obtained before surgery, and images were imported into imaging software. Optimum screw trajectories were selected for each vertebra, and drill guide templates were designed and fabricated with a 3-dimensional printing system. Drill guide templates were applied to cadaveric spine and clinical cases. Computed tomography imaging was performed after surgery, and planned and postoperative trajectories were compared to estimate the accuracy and safety of the drill guide templates. RESULTS: Twenty-two drill holes were made in cadaveric spinal specimens. All drill holes were completely located in the bone. The overall mean screw deviation was 0.88 ± 0.36 mm. In clinical cases, 29 screws were placed in thoracolumbar vertebrae. Most (89.6%) of these screws were placed without evidence of vertebral canal invasion. One (3.5%) screw perforated the bone structure. The overall mean screw deviation was 1.16 ± 0.56 mm. CONCLUSION: Drill guide templates were useful for accurate intraoperative screw navigation in thoracolumbar fixation in small dogs. CLINICAL SIGNIFICANCE: The use of drill guide templates can be considered as an aid to safety and accuracy of screw placement in canine thoracolumbar instabilities.
Asunto(s)
Tornillos Óseos , Vértebras Cervicales/cirugía , Enfermedades de los Perros/cirugía , Imagenología Tridimensional/veterinaria , Impresión Tridimensional , Cirugía Asistida por Computador/veterinaria , Animales , Cadáver , Perros , Femenino , Humanos , Inestabilidad de la Articulación/cirugía , Inestabilidad de la Articulación/veterinaria , Masculino , Periodo Posoperatorio , Estudios Prospectivos , Proyectos de Investigación , Tomografía Computarizada por Rayos X/métodosRESUMEN
BACKGROUND: Interleukin (IL)-33 has been implicated in the pathogenesis of canine atopic dermatitis, a Type 2 T helper cell (Th2)-associated disease. In humans, IL-33 mediates its biological effects through the receptor suppression of tumourigenicity 2 (ST2), which is preferentially expressed on Th2 cells. The effects of IL-33 on canine Th2 cells are unclear. HYPOTHESIS/OBJECTIVES: ST2 may be preferentially expressed on canine Th2 cells; IL-33 may induce the transcription of Th2 cytokines from these cells. ANIMALS: Three healthy dogs were used. METHODS: The transcription level of st2 was quantified in helper T cells, cytotoxic T cells and Th2 cells isolated from healthy dogs. The transcription levels of Th2 cytokines including il-4, il-5, il-13 and il-31 were quantified in Th2 cells stimulated with recombinant canine (rc) IL-33 and/or recombinant human (rh) IL-2. RESULTS: Transcription of st2 was the strongest in Th2 cells. Th2 cells also transcribed the genes for il-5 and il-13 after being stimulated with rcIL-33 and rhIL-2. CONCLUSIONS AND CLINICAL IMPORTANCE: These results indicate that canine Th2 cells activated by IL-33 enhance Th2-mediated inflammation through the production of IL-5 and IL-13.
Asunto(s)
Carcinogénesis/inmunología , Dermatitis Atópica/veterinaria , Enfermedades de los Perros/inmunología , Interleucina-33/genética , Receptores de Citocinas/genética , Células Th2/inmunología , Animales , Citocinas/genética , Citocinas/inmunología , Dermatitis Atópica/inmunología , Enfermedades de los Perros/metabolismo , Perros , Interleucina-13/genética , Interleucina-13/inmunología , Interleucina-33/inmunología , Interleucina-5/genética , Interleucina-5/inmunología , Receptores de Citocinas/antagonistas & inhibidores , Receptores de Citocinas/inmunología , Transcripción GenéticaRESUMEN
BACKGROUND: Previous studies indicate that tight junctions are involved in the pathogenesis of canine atopic dermatitis (cAD). An in vitro skin model is needed to elucidate the specific role of tight junctions in cAD. A 3D epidermal equivalent model using canine progenitor epidermal keratinocytes (CPEK) has been established; the expression of tight junctions within this model is uncharacterized. HYPOTHESIS/OBJECTIVES: To investigate the expression of tight junctions in the 3D epidermal equivalent. ANIMALS: Two normal laboratory beagle dogs served as donors of full-thickness skin biopsy samples for comparison to the in vitro model. METHODS: Immunohistochemical techniques were employed to investigate the expression of tight junctions including zonula occludens (ZO)-1 and claudin-1 in normal canine skin, and in the CPEK 3D epidermal equivalent. RESULTS: Results demonstrated the expression of ZO-1 and claudin-1 in the CPEK 3D epidermal equivalent, with staining patterns that were similar to those in normal canine skin. CONCLUSIONS AND CLINICAL IMPORTANCE: The CPEK 3D epidermal equivalent has the potential to be a suitable in vitro research tool for clarifying the specific role of tight junctions in cAD.
RESUMEN
BACKGROUND: In humans, interleukin (IL)-33 plays a critical role in the enhancement of allergic skin inflammation. However, it currently remains unclear whether IL-33 is involved in the pathogenesis of canine atopic dermatitis (cAD). OBJECTIVES: To examine the expression of IL-33 in chronic lesional skin of cAD. ANIMALS: Eight dogs with spontaneous cAD and five healthy dogs were used. METHODS: The transcription of il-33 in chronic lesional skin of cAD was quantified by quantitative reverse transcription PCR. The expression of IL-33 was evaluated immunohistochemically using an anti-human IL-33 monoclonal antibody with cross-reactivity to canine IL-33. RESULTS: The transcription levels of il-33 in chronic lesional skin of cAD were significantly higher than those in normal skin of healthy dogs. Keratinocytes were a major cellular source of IL-33 production in chronic lesional skin of cAD. CONCLUSIONS AND CLINICAL IMPORTANCE: The results indicate that IL-33 is involved in chronic lesional skin of cAD.
Asunto(s)
Dermatitis Atópica/veterinaria , Enfermedades de los Perros/inmunología , Interleucina-33/metabolismo , Animales , Dermatitis Atópica/inmunología , Dermatitis Atópica/metabolismo , Dermatitis Atópica/patología , Enfermedades de los Perros/metabolismo , Enfermedades de los Perros/patología , Perros , Regulación de la Expresión Génica/fisiología , Interleucina-33/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , PielRESUMEN
BACKGROUND: In canine epitheliotropic cutaneous T-cell lymphoma (ECTCL), neoplastic cells cause skin lesions and potentially metastasize to lymph nodes, blood and other organs. Murine models are potentially valuable for elucidating the molecular mechanisms responsible for regulation of ECTCL cell migration. HYPOTHESIS/OBJECTIVES: To describe a phenotype of mice xenografted with canine ECTCL cells (EO-1 cells). ANIMALS: Four NOD.CB17-Prkdcscid /J (NOD SCID) mice were used. METHODS AND MATERIALS: EO-1 cells were subcutaneously xenografted into NOD SCID mice. After four weeks, the development of tumour lesions in skin and other organs was investigated. RESULTS: Mice developed skin lesions with metastasis to the lymph nodes, spleen, lung, blood and liver. CONCLUSIONS AND CLINICAL IMPORTANCE: Mice xenografted with EO-1 cells may be useful for studying the pathogenesis of canine ECTCL.
Asunto(s)
Linfoma Cutáneo de Células T/veterinaria , Neoplasias Cutáneas/veterinaria , Animales , Modelos Animales de Enfermedad , Perros , Femenino , Xenoinjertos , Linfoma Cutáneo de Células T/patología , Ratones , Ratones Endogámicos NOD , Ratones SCID , Trasplante de Neoplasias/veterinaria , Piel/patología , Neoplasias Cutáneas/patologíaRESUMEN
BACKGROUND: Dysfunction of the physical and chemical barriers of the skin may play roles in the pathogenesis of atopic dermatitis (AD) by facilitating penetration of antigens through the skin and consequently evoking aberrant immune reactions. It is now emerging that keratinocytes are actively involved in cutaneous immune reactions by producing various soluble factors initiated by inflammatory stimuli, including mechanical injury or activation of Toll-like receptors and protease-activated receptors. Among the soluble factors, keratinocyte-derived cytokines and chemokines skew Type 2 helper T (Th2) cell-dominant immune reactions, with the recruitment of Th2 cells. OBJECTIVE: To review the roles of keratinocyte-derived cytokines and chemokines in the pathogenesis of AD in humans and dogs. CONCLUSION AND CLINICAL IMPORTANCE: Keratinocyte-derived cytokines such as thymus and activation-regulated chemokine, granulocyte-macrophage colony stimulating factor, thymic stromal lymphopoietin and interleukin-33 are involved in the pathogenesis of human AD and possibly in canine AD. These cytokines and chemokines may possibly be used as subjective clinical markers and therapeutic targets for both human and canine AD.
Asunto(s)
Quimiocinas/fisiología , Citocinas/fisiología , Dermatitis Atópica/etiología , Enfermedades de los Perros/etiología , Queratinocitos/fisiología , Animales , Dermatitis Atópica/inmunología , Dermatitis Atópica/veterinaria , Enfermedades de los Perros/inmunología , Perros , Humanos , Piel/inmunologíaRESUMEN
BACKGROUND: Colonization, overgrowth and subsequent infection by Staphylococcus spp. is frequently observed in canine atopic dermatitis (CAD), where it contributes to the intensity of cutaneous inflammation. The mechanisms by which staphylococci contribute to the pathogenesis of CAD are unclear. Studies suggest that thymic stromal lymphopoietin (TSLP), a cytokine induced by a cell wall component of Staphylococcus spp., may play a critical role in Th2 responses including the pathogenesis of CAD. HYPOTHESIS/OBJECTIVE: To determine if synthetic triacylated lipopeptide (TLR1/2 ligand), a cell wall component of Staphylococcus spp., induces the transcription of TSLP via TLR2 in canine keratinocytes. METHODS: Transcription of TSLP was quantified in a canine keratinocyte cell line after stimulation with synthetic triacylated lipopeptide, and again after inhibition of TLR2 by a targeted small interfering RNA. RESULTS: The transcription of TSLP was enhanced 6 h after stimulation with the synthetic triacylated lipopeptide; it was completely suppressed by knockdown of TLR2. CONCLUSIONS AND CLINICAL IMPORTANCE: The results demonstrated that a synthetic cell wall component of Staphylococcus spp. induced transcription of TSLP via TLR2 in canine keratinocytes. Additional studies will be required to investigate whether Staphylococcus spp. contributes to Th2 responses in CAD through TLR2-mediated TSLP production.
Asunto(s)
Citocinas/metabolismo , Dermatitis Atópica/veterinaria , Enfermedades de los Perros/metabolismo , Queratinocitos/metabolismo , Staphylococcus , Receptor Toll-Like 2/metabolismo , Animales , Citocinas/genética , Dermatitis Atópica/complicaciones , Dermatitis Atópica/metabolismo , Enfermedades de los Perros/etiología , Enfermedades de los Perros/patología , Perros , Regulación de la Expresión Génica/inmunología , Hipersensibilidad/veterinaria , Inflamación/metabolismo , Inflamación/veterinaria , Infecciones Cutáneas Estafilocócicas/complicaciones , Infecciones Cutáneas Estafilocócicas/metabolismo , Infecciones Cutáneas Estafilocócicas/veterinaria , Receptor Toll-Like 2/genética , Linfopoyetina del Estroma TímicoRESUMEN
BACKGROUND: Pro-inflammatory cytokines and chemokines produced by activated keratinocytes play an important role in the pathogenesis of canine atopic dermatitis (AD) as well as human AD. Recent studies suggest that keratinocytes activated by IL-17A are involved in the pathogenesis of human AD. However, the role of IL-17A in canine keratinocytes is poorly understood. HYPOTHESIS/OBJECTIVES: Interleukin-17A would induce the transcription of pro-inflammatory cytokines and chemokines in canine keratinocytes. METHODS: The transcription levels of pro-inflammatory cytokines and chemokines were quantified in a canine keratinocyte cell line stimulated with recombinant canine (rc) IL-17A. RESULTS: The transcription of GM-CSF, S100A8, IL-8 and IL-19 in cultured keratinocytes was significantly enhanced at 24 h after stimulation with rcIL-17A. CONCLUSIONS AND CLINICAL IMPORTANCE: Keratinocytes activated by IL-17A have the ability to produce various pro-inflammatory cytokines and chemokines, suggesting that IL-17A may play a central role of the development of Th2-associated inflammation in canine AD.
Asunto(s)
Citocinas/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Interleucina-17/farmacología , Queratinocitos/efectos de los fármacos , Transcripción Genética/efectos de los fármacos , Animales , Citocinas/genética , Perros , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinariaRESUMEN
BACKGROUND: Previous studies reported the involvement of CC chemokine receptor 4 (CCR4)-positive CD4(+) cells in the pathogenesis of canine atopic dermatitis. In humans, CCR4 is selectively expressed on type 2 helper T (Th2) cells; however, a subset of canine CCR4(+) helper T cells has not been determined. HYPOTHESIS/OBJECTIVES: To characterize the transcription profile of CCR4(+) CD4(+) lymphocytes isolated from the peripheral blood of healthy dogs. ANIMALS: Three healthy dogs were used. METHODS: The transcription levels of type 1 helper T (Th1) and Th2 cytokines in CCR4(+) CD4(+) and CCR4(-) CD4(+) lymphocytes isolated from healthy dogs were quantified by real-time RT-PCR. RESULTS: The CCR4(+) CD4(+) lymphocytes preferentially transcribed Th2 cytokines, such as interleukin-4 and interleukin-13, but not Th1 cytokines, such as interferon-γ. CONCLUSIONS AND CLINICAL IMPORTANCE: CCR4 can be used as a specific marker of Th2 cells for elucidation of the pathogenesis or the establishment of novel therapeutics in canine Th2-associated diseases, such as canine atopic dermatitis.
Asunto(s)
Linfocitos T CD4-Positivos/metabolismo , Citocinas/metabolismo , Regulación de la Expresión Génica/fisiología , Receptores CCR4 , Animales , Perros , Femenino , Masculino , TranscriptomaRESUMEN
BACKGROUND: A previous study demonstrated that the cysteine protease of Dermatophagoides farinae induced production of granulocyte-macrophage colony-stimulating factor (GM-CSF) in a canine epidermal keratinocyte progenitor cell line (CPEK); however, the molecular mechanism has not been elucidated. HYPOTHESIS/OBJECTIVES: Given that the transcription of GM-CSF mRNA in human lymphocytes is mainly regulated by the nuclear factor of activated T cells (NFAT), it is hypothesized that NFAT also contributes to GM-CSF production in canine keratinocytes stimulated with a cysteine protease. METHODS: Nuclear translocation of NFAT was evaluated in CPEK cells in the absence or presence of the cysteine protease papain. We also investigated whether blockade of NFAT could inhibit GM-CSF production. RESULTS: Papain-induced nuclear translocation of NFAT, producing GM-CSF, was partly inhibited by ciclosporin. CONCLUSIONS AND CLINICAL IMPORTANCE: The results suggest that GM-CSF production mediated by the cysteine protease is regulated not only by NFAT but also by unknown signalling pathways in canine keratinocytes.
Asunto(s)
Perros , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Queratinocitos/efectos de los fármacos , Queratinocitos/metabolismo , Factores de Transcripción NFATC/metabolismo , Animales , Línea Celular , Ciclosporina/farmacología , Inhibidores de Cisteína Proteinasa/administración & dosificación , Inhibidores de Cisteína Proteinasa/farmacología , Relación Dosis-Respuesta a Droga , Regulación de la Expresión Génica/efectos de los fármacos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Factores de Transcripción NFATC/genética , Papaína/administración & dosificación , Papaína/farmacología , Células Madre/efectos de los fármacos , Células Madre/metabolismoRESUMEN
BACKGROUND: Mycosis fungoides (MF) is the most common form of canine epitheliotropic cutaneous lymphoma, which is characterized by the accumulation of neoplastic CD8(+) T cells. Given that multifocal skin lesions are commonly seen in MF, neoplastic lymphocytes may actively migrate into the blood circulation. HYPOTHESIS/OBJECTIVES: Cytotoxic T cells with a skin-homing phenotype could be increased in the blood circulation of dogs with MF. ANIMALS: Ten dogs with MF and 10 age-matched healthy dogs were included. METHODS: The transcription levels of chemokine receptors, cytokines and cytotoxic markers in peripheral blood of dogs with MF were quantified by real-time RT-PCR. RESULTS: The dogs with MF had lower transcription levels of chemokine receptors associated with skin homing (CCR4), epitheliotropism (CXCR3), lymph node homing (CCR7), a type-1 cytokine (LT-α) and cytotoxic markers (perforin and granzyme B) in the circulation than healthy control dogs (P < 0.05). CONCLUSIONS AND CLINICAL IMPORTANCE: The present results suggest that the number of peripheral cytotoxic T cells with a skin-homing phenotype could be decreased in the peripheral blood of dogs with MF, which might be due to the sequestration of cytotoxic T cells in the lesional skin.
Asunto(s)
Biomarcadores de Tumor/sangre , Citocinas/metabolismo , Enfermedades de los Perros/metabolismo , Micosis Fungoide/veterinaria , Receptores de Quimiocina/metabolismo , Transcriptoma , Animales , Estudios de Casos y Controles , Citocinas/genética , Enfermedades de los Perros/sangre , Perros , Regulación Neoplásica de la Expresión Génica , Micosis Fungoide/genética , Micosis Fungoide/metabolismo , Receptores de Quimiocina/genéticaRESUMEN
Background: Precursor-targeted immune-mediated anemia (PIMA) has been described in dogs presenting with nonregenerative anemia and evidence of ineffective erythropoiesis. Although it has been suggested that its occurrence may be related to the immune targeting of erythroid precursors, this pathogenesis has not been established. PIMA is mainly treated with glucocorticoids, and in cases where glucocorticoids alone are not effective, immunosuppressants are also used as combination therapy. However, not all cases of PIMA go into remission after these treatments. Case Description: Two dogs with severe nonregenerative anemia diagnosed as PIMA based on the results of clinical pathological examinations, including bone marrow examination, were treated with whole-blood transfusion and immunosuppressive doses of prednisolone, mycophenolate mofetil, and cyclosporine. However, these treatments failed to achieve remission of PIMA. Therefore, concomitant administration of oclacitinib, which is a Janus kinase-1 inhibitor that has been applied recently to the treatment of immune-mediated diseases, was performed; this combined regimen improved the anemia and achieved complete remission of PIMA. Conclusion: Oclacitinib may be an option for the treatment of PIMA in dogs failing to achieve remission with conventional immunosuppressive therapy.
Asunto(s)
Anemia , Enfermedades de los Perros , Perros , Animales , Ciclosporina/uso terapéutico , Prednisolona/uso terapéutico , Yoduro de Potasio/uso terapéutico , Inmunosupresores/uso terapéutico , Anemia/tratamiento farmacológico , Anemia/etiología , Anemia/veterinaria , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de los Perros/patologíaRESUMEN
Aims: C16 monounsaturated fatty acid (C16:1) show antibacterial activity against Staphylococcus aureus, a pathogen associated with various diseases such as atopic dermatitis and bacteremia, while the compound does not exhibit antibacterial activity against Staphylococcus epidermidis, an epidermal commensal that inhibits the growth of S. aureus. In this study, we aimed to find bifidobacterial strains with the ability to produce C16:1 and to find a practical manner to utilize C16:1-producing strains in industry. Methods: Various Bifidobacterium strains were screened for their content of C16:1. The chemical identity of C16:1 produced by a selected strain was analyzed by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS). Medium components that affect the C16:1 content of the selected strain were investigated. Antibacterial activity against staphylococci was compared between the authentic C16:1 isomers and total fatty acids (TFA) extracted from the selected strain. Results: B. adolescentis 12451, B. adolescentis 12-111, B. boum JCM 1211, and Bifidobacterium sp. JCM 7042 showed high C16:1 content among the tested strains. TFA extracted from Bifidobacterium sp. JCM 7042 contained C16:1 at 2.3% as the fatty acid constituent (2.4 mg/L of broth). Through GC-MS and LC-MS analyses, the C16:1 synthesized by Bifidobacterium sp. JCM 7042 was identified as 7-cis-hexadecenoic acid (7-cis-C16:1). The authentic 7-cis-C16:1 showed strong and selective antibacterial activity against S. aureus, similar to 6-cis-C16:1, with a minimum inhibitory concentration (MIC) of < 10 µg/mL. Components that increase C16:1 productivity were not found in the MRS and TOS media; however, Tween 80 was shown to considerably reduce the C16:1 ratio in TFA. Antibacterial activity against S. aureus was observed when the TFA extracted from Bifidobacterium sp. JCM 7042 contained high level of 7-cis-C16:1 (6.1% in TFA) but not when it contained low level of 7-cis-C16:1 (0.1% in TFA). Conclusion: The fatty acid, 7-cis-C16:1, which can selectively inhibit the S. aureus growth, is accumulated in TFA of several bifidobacteria. The TFA extracted from cultured cells of Bifidobacterium sp. JCM 7042 demonstrated antibacterial activity. From a practical viewpoint, our findings are important for developing an efficient method to produce novel skin care cosmetics, functional dairy foods, and other commodities.
RESUMEN
C-C chemokine receptor 7 (CCR7) contributes to cell homing to lymph nodes (LNs). Recent studies reported that CCR7 is also expressed in tumor cells, which correlates with LN metastasis in various cancers. However, the expression of CCR7 in tumor cells is unknown in dogs due to the lack of appropriate antibodies. In the present study, a fusion protein of C-C chemokine ligand 19 (CCL19) was employed as an alternative method to CCR7 antibodies. The fusion CCL19 protein specifically detected CCR7 expressed in canine lymphoma cell lines, which showed active chemotaxis to both canine and mouse ligands. The present study will help further research on the involvement of canine CCR7 in LN metastasis.
Asunto(s)
Enfermedades de los Perros , Linfoma , Enfermedades de los Roedores , Animales , Línea Celular , Perros , Ganglios Linfáticos , Linfoma/veterinaria , Ratones , Receptores CCR7RESUMEN
The objective of this study was to evaluate the feasibility and clinical outcomes of microendoscopic dorsal laminectomy for multi-level cervical intervertebral disc protrusions in dogs. Eight client-owned dogs diagnosed with multi-level cervical intervertebral disc protrusions using computed tomography (CT) and magnetic resonance imaging (MRI) were included in this retrospective case series. Microendoscopic dorsal laminectomies (MEL) were performed with an integrated endoscopic system to the cranial and caudal vertebrae of the affected intervertebral joints. Pre- and post-operative neurological status, operation time, intra-operative complications, and postoperative complications were reviewed. Post-operative CT images were obtained to measure the dimensions of laminectomy and compared to those of planned laminectomy. Full endoscopic procedures were feasible in 7 dogs (87.5%) and the laminectomy dimensions were in agreement with pre-operative planning. In all dogs, major intra- and postoperative complications did not occur. Conversion to open surgery was required in one case. Short-term postoperative clinical deterioration was found in two dogs. Long-term clinical outcomes were good and comparable to those reported in previous studies of open dorsal laminectomies. MEL is a promising minimally invasive approach to multi-level cervical dorsal laminectomy for intervertebral disc protrusions. This technique may improve postoperative discomfort compared to the open approach. Further studies are needed to directly compare outcomes between these two approaches.
RESUMEN
Canine degenerative myelopathy (DM) is a progressive neurodegenerative disease of the spinal cord. The diagnosis is based on the observation of clinical signs, genetic testing, and exclusion of other spinal cord diseases, and a definitive diagnosis of DM can only be confirmed by postmortem histopathological findings. The aim of this study was to investigate the diagnostic ability of diffusion tensor imaging (DTI) for DM. Eight DM-affected Pembroke Welsh Corgis, thirteen dogs with thoracolumbar intervertebral disk herniation (IVDH), and six healthy control dogs were included. All dogs were scanned using a 3.0-T MRI system. Apparent diffusion coefficient (ADC) and fractional anisotropy (FA) values were calculated for each intervertebral disk level slice between T8-T9 and L2-L3 intervertebral disk levels, and the entire area of the thoracolumbar spinal cord between T8-T9 and L2-L3 intervertebral disk levels (T8-L3 region). The ADC and FA values of the T8-L3 region were significantly lower in the DM group than in the IVDH group. The ADC values for the T8-L3 region had a moderate negative correlation with clinical duration (rs= -0.723, P=0.043); however, the FA values of other intervertebral disk levels and T8-L3 region had no correlation with clinical durations. The measurement of DTI indices can be used to quantitatively assess neurodegeneration and may have diagnostic value for DM. In particular, the ADC value of the T8-L3 region may aid in making a non-invasive premortem diagnosis of DM.