Interleukin 10 and transforming growth factor‐beta 1 plasma levels in atopic dogs before and during immunotherapy

Background: Human studies suggest that the cytokines, interleukin 10 (IL-10) and transforming growth factor-beta 1 (TGF-ß1) may play an important role in allergen-specific immunotherapy (ASIT). However, there is little known about the function of these cytokines in atopic dogs. This study compared the plasma levels of IL-10 and TGF-ß1 in atopic and control dogs and investigated their changes during different ASIT approaches. Methods: A total of 54 atopic and 32 control dogs were included. Immunotherapy was performed in 30 atopic dogs. The dogs undergoing immunotherapy were allocated to four groups of different ASIT approaches (namely subcutaneous, intralymphatic, sublingual ASIT and subcutaneous ASIT with recombinant allergens). Blood samples were collected at four timepoints throughout the one year of ASIT. Canine atopic dermatitis extent and severity index, pruritus visual analogue scale and medication score were recorded at each timepoint. Commercially available ELISA kits were used to quantify IL-10 and TGF-ß1 in plasma. Results: There was no significant difference in IL-10 and TGF-ß1 between atopic and control dogs. The IL-10 levels were significantly increased in the intralymphatic group at the end of the study. No significant differences were found in the other groups for both IL-10 and TGF-ß1. Conclusion: The findings of this work suggest that IL-10 and TGF-ß1 cannot be used to monitor the course of the disease during ASIT

A novel therapeutic diet can significantly reduce the medication score and pruritus of dogs with atopic dermatitis during a nine‐month controlled study

Background: Canine atopic dermatitis (cAD) is a common chronic relapsing pruritic skin disease for which management commonly relies on life-long use of immunomodulatory drugs. A number of the medications used are associated with adverse effects and the potential for complications during long-term use. Hypothesis: The goal of the study was to determine if a complete and balanced diet formulated for therapeutic benefit could contribute towards management of cAD. We hypothesised that the diet would reduce pruritus while also reducing the requirement for medication during the study period. Animals, materials and methods: Forty privately owned dogs, having undergone a comprehensive diagnosis for cAD, were randomly allocated to two groups, each group being fed one of two diets (test or control) for up to nine months. We assessed pruritus, Canine Atopic Dermatitis Extent and Severity Index-(4th iteration) and medication score, the latter reflecting the medication required to maintain a satisfactory quality of life for the animal. Results: Both diets were well-accepted and -tolerated. There was a significant improvement in the pruritus score after three months of feeding the therapeutic diet (P = 0.0001). No such improvement was observed at any time point in the group of dogs given the control diet. There was a reduced drug requirement for dogs receiving the therapeutic diet after three months (P = 0.058), and that decrease was significant at six months (P = 0.021) and nine months (P = 0.018). No improvement was seen at any time point in the control group. Conclusion: The results suggest that a novel therapeutic diet can assist in the management of cAD by helping to control pruritus and reducing reliance on medication

Independent DSG4 frameshift variants in cats with hair shaft dystrophy

Investigations of hereditary phenotypes in spontaneous mutants may help to better understand the physiological functions of the altered genes. We investigated two unrelated domestic shorthair cats with bulbous swellings of the hair shafts. The clinical, histopathological, and ultrastructural features were similar to those in mice with lanceolate hair phenotype caused by loss-of-function variants in Dsg4 encoding desmoglein 4. We sequenced the genomes from both affected cats and compared the data of each affected cat to 61 control genomes. A search for private homozygous variants in the DSG4 candidate gene revealed independent frameshift variants in each case, c.76del or p.Ile26fsLeu*4 in case no. 1 and c.1777del or p.His593Thrfs*23 in case no. 2. DSG4 is a transmembrane glycoprotein located primarily in the extracellular part of desmosomes, a complex of adhesion molecules responsible for connecting the keratin intermediate filaments of neighbouring epithelial cells. Desmosomes are essential for normal hair shaft formation. Both identified DSG4 variants in the affected cats lead to premature stop codons and truncate major parts of the open-reading frame. We assume that this leads to a complete loss of DSG4 function, resulting in an incorrect formation of the desmosomes and causing the development of defective hair shafts. Together with the knowledge on the effects of DSG4 variants in other species, our data suggest that the identified DSG4 variants cause the hair shaft dystrophy. To the best of our knowledge, this study represents the first report of pathogenic DSG4 variants in domestic animals

Comparison of the Gut Microbiome between Atopic and Healthy Dogs—Preliminary Data

Human studies show that in addition to skin barrier and immune cell dysfunction, both the cutaneous and the gut microbiota can influence the pathogenesis of atopic diseases. There is currently no data on the gut-skin axis in allergic canines. Therefore, the aim of this study was to assess the bacterial diversity and composition of the gut microbiome in dogs with atopic dermatitis (AD). Stool samples from adult beagle dogs (n = 3) with spontaneous AD and a healthy control group (n = 4) were collected at Days 0 and 30. After the first sampling, allergic dogs were orally dosed on a daily basis with oclacitinib for 30 days, and then re-sampled. Sequencing of the V3–V4 region of the 16S rRNA gene was performed on the Illumina MiSeq platform and the data were analyzed using QIIME2. The atopic dogs had a significantly lower gut microbiota alpha-diversity than healthy dogs (p = 0.033). In healthy dogs, a higher abundance of the families Lachnospiraceae (p = 0.0006), Anaerovoracaceae (p = 0.006) and Oscillospiraceae (p = 0.021) and genera Lachnospira (p = 0.022), Ruminococcustorques group (p = 0.0001), Fusobacterium (p = 0.022) and Fecalibacterium (p = 0.045) was seen, when compared to allergic dogs. The abundance of Conchiformibius (p = 0.01), Catenibacterium spp. (p = 0.007), Ruminococcus gnavus group (p = 0.0574) and Megamonas (p = 0.0102) were higher in allergic dogs. The differences in alpha-diversity and on the compositional level remained the same after 1 month, adding to the robustness of the data. Additionally, we could also show that a 4-week treatment course with oclacitinib was not associated with changes in the gut microbiota diversity and composition in atopic dogs. This study suggests that alterations in the gut microbiota diversity and composition may be associated with canine AD. Large-scale studies preferably associated to a multi-omics approach and interventions targeting the gut microbiota are needed to confirm these results

Independent DSG4 frameshift variants in cats with hair shaft dystrophy

Investigations of hereditary phenotypes in spontaneous mutants may help to better understand the physiological functions of the altered genes. We investigated two unrelated domestic shorthair cats with bulbous swellings of the hair shafts. The clinical, histopathological, and ultrastructural features were similar to those in mice with lanceolate hair phenotype caused by loss-of-function variants in Dsg4 encoding desmoglein 4. We sequenced the genomes from both affected cats and compared the data of each affected cat to 61 control genomes. A search for private homozygous variants in the DSG4 candidate gene revealed independent frameshift variants in each case, c.76del or p.Ile26fsLeu*4 in case no. 1 and c.1777del or p.His593Thrfs*23 in case no. 2. DSG4 is a transmembrane glycoprotein located primarily in the extracellular part of desmosomes, a complex of adhesion molecules responsible for connecting the keratin intermediate filaments of neighbouring epithelial cells. Desmosomes are essential for normal hair shaft formation. Both identified DSG4 variants in the affected cats lead to premature stop codons and truncate major parts of the open-reading frame. We assume that this leads to a complete loss of DSG4 function, resulting in an incorrect formation of the desmosomes and causing the development of defective hair shafts. Together with the knowledge on the effects of DSG4 variants in other species, our data suggest that the identified DSG4 variants cause the hair shaft dystrophy. To the best of our knowledge, this study represents the first report of pathogenic DSG4 variants in domestic animals

Independent COL17A1 Variants in Cats with Junctional Epidermolysis Bullosa.

Epidermolysis bullosa (EB), characterized by defective adhesion of the epidermis to the dermis, is a heterogeneous disease with many subtypes in human patients and domestic animals. We investigated two unrelated cats with recurring erosions and ulcers on ear pinnae, oral mucosa, and paw pads that were suggestive of EB. Histopathology confirmed the diagnosis of EB in both cats. Case 1 was severe and had to be euthanized at 5 months of age. Case 2 had a milder course and was alive at 11 years of age at the time of writing. Whole genome sequencing of both affected cats revealed independent homozygous variants in COL17A1 encoding the collagen type XVII alpha 1 chain. Loss of function variants in COL17A1 lead to junctional epidermolysis bullosa (JEB) in human patients. The identified splice site variant in case 1, c.3019+1del, was predicted to lead to a complete deficiency in collagen type XVII. Case 2 had a splice region variant, c.769+5G>A. Assessment of the functional impact of this variant on the transcript level demonstrated partial aberrant splicing with residual expression of wildtype transcript. Thus, the molecular analyses provided a plausible explanation of the difference in clinical severity between the two cases and allowed the refinement of the diagnosis in the affected cats to JEB. This study highlights the complexity of EB in animals and contributes to a better understanding of the genotype-phenotype correlation in COL17A1-related JEB

Bienen- und Wespengiftallergien bei Hunden – Von der Akutbehandlung bis zur Desensibilisierung

Insektengifte zählen zu den häufigsten Allergieauslösern des Hundes. Das klinische Bild reicht von lokalen Reaktionen über allergische Allgemeinreaktionen bis hin zu Intoxikationen. Der Beitrag gibt wichtige Informationen über die diagnostische Aufarbeitung, die Akutbehandlung und präventive Maßnahmen

Circulating CD4(+)CD25(+)Foxp3(+) T regulatory cell levels in an experimental model of canine atopic dermatitis

BACKGROUND: The pathogenesis of canine atopic dermatitis (cAD) is characterized immunologically by an imbalanced T-cell response. Mechanisms of immune regulation in cAD have not yet been completely elucidated. OBJECTIVES: To investigate peripheral blood T regulatory (Treg) cells and their associated cytokines (TGF-β and IL-10) in an experimental model of cAD. ANIMALS: Eight beagle dogs that were initially naïve and subsequently sensitized to house dust mites (HDM). METHODS AND MATERIALS: T regulatory cell phenotyping was performed by flow-cytometric analysis on peripheral blood; serum cytokine levels were measured by ELISA, before sensitization and after challenge with HDM allergens. Additionally, clinical scores and allergen-specific IgE were determined. RESULTS: After challenge of sensitized dogs to HDM allergen, a significant increase of Treg cells and simultaneous decrease in the serum TGF-β were observed. However, in most dogs, serum IL-10 values were below the detection limit. Treg cell proportions before sensitization were significantly negatively correlated with the HDM-specific IgE levels and clinical scores after induction of AD signs. CONCLUSION AND CLINICAL IMPORTANCE: The results confirm that Treg responses are involved in the pathogenesis of an experimental model cAD. Further investigations are required to clarify the precise immune modulating function of canine Treg cells and their interplay with other immune cell types