Open trial of recombinant Der f 2 pullulan-conjugated immunotherapy in cats.

BACKGROUND: Allermmune HDM (Zenoaq) is a recombinant Dermatophagoides farinae 2 (Der f 2) pullulan-based immunotherapy vaccine whose efficacy on house dust mite allergic dogs has been demonstrated. There is no published information on its use in cats. OBJECTIVES: The objective of the study was to evaluate the safety and short-term effects of Allermmune HDM in Dermatophagoides farinae (Df)-sensitised cats. MATERIALS AND METHODS: Eleven cats diagnosed with atopic skin syndrome received Allermmune weekly for six weeks then monthly for three months (total duration 18 weeks). On Weeks 0, 6 and 18 clinical lesions were assessed by the Feline Dermatitis Extent and Severity Index (FEDESI); owners assessed pruritus with a 10-cm Visual Analog Scale (pVAS). Concurrent medication use was recorded. The allergen-specific immunoglobulin (Ig)E were measured before study inclusion with a commercial serological assay. RESULTS: There were no evident adverse effects. FEDESI and pVAS improved significantly after six weeks (p = 0.001 and p = 0.01, respectively). The pretreatment Df-specific IgE levels were significantly higher in the cats with improved clinical scores than in the cats with no clinical score change (p = 0.009). CONCLUSIONS AND CLINICAL RELEVANCE: Allermmune HDM may be safe in cats and has the potential to alleviate signs of atopic skin syndrome. Allergen-specific IgE levels may represent an efficacy marker. Controlled studies of longer duration and larger sample size are worth pursuing.

Identification of cross-reactive allergens between the Dermatophagoides farinae house dust mite and the Toxocara canis nematode in dogs with suspected allergies.

BACKGROUND: Immunoglobulin (Ig)E cross-reactivity has been shown between Dermatophagoides farinae (Df; house dust mite) and the nematode Toxocara canis (Tc), yet its allergen basis is unknown. OBJECTIVES: To identify the Df allergens IgE-cross-reactive with those of Tc. ANIMALS: Archived sera from 73 dogs with suspected allergy sensitised to Df. MATERIALS AND METHODS: We performed a combination of Pet Allergy Xplorer (PAX) and enzyme-linked immunosorbent assay (ELISA) inhibitions with excretory-secretory and somatic (i.e. nematode body) extracts of Tc or recombinant Tc tropomyosin on coats of Df, Der f 15 and Zen-1 (ELISA) or PAX allergens. RESULTS: The ELISA and PAX inhibitions established that there is mutual yet variable cross-reactivity between the Tc excretory-secretory extract, purified Der f 15 and purified Zen-1. This cross-reactivity is likely to involve cross-reactive glycans, as there is no inhibition between the Tc excretory-secretory extract and recombinant Der f 15 without its predicted natural O-glycans. We also confirmed a heterogeneous cross-reactivity between the somatic Tc extract and Der p 11 (paramyosin), as well as between the recombinant Toxo c 3 and Der p 10 tropomyosins. The cross-reactivity among tropomyosins and paramyosins is likely to involve peptidic epitopes, as these recombinant allergens are not glycosylated. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs with suspected allergies, the cross-reactivity between Tc and Df for dogs is complex and heterogeneous. Some of the cross-reactive IgE recognises shared glycans on Der f 15 and Zen-1, while some targets peptidic epitopes on shared paramyosins and tropomyosins. We do not exclude that additional cross-reactive allergens between Df and Tc also might exist.

Rapid induction of allergen-blocking IgG in dogs vaccinated with plant-based, Der f 2-expressing bioparticles.

BACKGROUND: Allergen-carrying virus-like particles are effective and safe means of allergen immunotherapy (AIT) in rodent models. OBJECTIVE: To study the development of allergen-blocking immunoglobulin (Ig)G in dogs injected with Der f 2-carrying enveloped plant-based bioparticles (eBPs). MATERIALS AND METHODS: Laboratory beagle dogs were injected intradermally (ID) or subcutaneously (SC) with Der f 2-eBP three times at 2-week intervals. A basophil mediator release assay was used to compare the reactivity of Der f 2-eBPs to that of recombinant Der f 2. Allergen-specific IgG serum levels were determined by immunoblotting and ELISA. The allergen-blocking potential of postvaccination IgG was assessed by Pet Allergy Xplorer (PAX) macroarray and basophil mediator release inhibition assays. RESULTS: The amount of Der f 2 eBPs needed to induce basophil activation was 1000-fold higher than that of the soluble natural allergen. In both immunisation groups, eBP injections caused no adverse events and induced Der f 2-specific IgG, first detected on Day (D)14 and peaking on D41. The co-incubation of sera with a Der f 2-IgE-rich canine serum pool resulted in a mean PAX inhibition of 70% (ID) to 80% (SC) on D41. For both groups, the inhibition of basophil mediator release reached 75% on D28 and D41. The percentage inhibition of PAX and mediator release correlated significantly with Der f 2 IgG levels. CONCLUSION AND CLINICAL RELEVANCE: Intradermal and subcutaneous injections of Der f 2-eBPs were safe and increased Der f 2-specific IgG. The clinical benefit of immunotherapy will be evaluated in future trials enrolling atopic dogs allergic to house dust mites.

An international seroprevalence survey of the IgE sensitisation to the Dermatophagoides farinae house dust mite and two of its major allergens (Der f 2, Zen 1) in atopic dogs.

BACKGROUND: Dogs with atopic dermatitis are often immunoglobulin (Ig)E-sensitised to Dermatophagoides farinae (Df) house dust mites, yet limited data exist on the sensitisation rates to the individual Df allergens, Der f 2 and Zen 1. OBJECTIVES: To determine the IgE sensitisation rates to Df, Der f 2 and Zen 1 in atopic dogs from geographically diverse countries. ANIMALS: Serum was collected from 32 laboratory dogs in Japan, and 837 atopic dogs from 11 countries from five continents: Asia (Japan, Thailand, Taiwan), Europe (Italy, Latvia, the Netherlands, UK), North America (USA), South America (Argentina, Brazil) and Africa (South Africa). METHODS AND MATERIALS: We determined Df-, Der f 2- and Zen 1-specific IgE levels by ELISA. Correlations between the IgE values for these three allergens were calculated. RESULTS: The IgE seropositivity rates for Df varied between 74% (Argentina) and 100% (the Netherlands, Thailand, South Africa), those for Der f 2 between 12% (Argentina) and 88% (South Africa), and for Zen 1 between 70% (Argentina) and 100% (the Netherlands). Apart from the especially low seropositivity rate for Der f 2-specific IgE in Argentina, the percentage of IgE sensitisation varied little between countries. There was significant correlation between the IgE levels to these three allergens which was highest between Df and Zen 1, and lowest between Zen 1 and Der f 2. CONCLUSIONS AND CLINICAL RELEVANCE: The IgE sensitisation to Df is geographically widespread. Der f 2 and Zen 1 are major allergens for dogs in almost all countries where this was evaluated.

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.

Phaeohyphomycosis caused by Phialophora americana in a dog.

Phaeohyphomycosis was diagnosed in a 6-year-old, male castrated Dachshund on immunosuppressive treatment. The fungus was identified by culture and PCR as Phialophora americana. This is the first reported case of infection with this pathogen in a dog. The infection was successfully managed medically, without surgical intervention.

Une phaéohyphomycose a été diagnostiquée chez un teckel mâle castré de 6 ans sous traitement immunosuppresseur. Le champignon a été identifié par culture et PCR comme Phialophora americana. Il s'agit du premier cas rapporté d'infection par cet agent pathogène chez un chien. L'infection a été prise en charge médicalement avec succès, sans intervention chirurgicale.

Se diagnosticó feohifomicosis en un macho de Teckel castrado de 6 años en tratamiento inmunosupresor. El hongo fue identificado por cultivo y PCR como Phialophora americana. Este es el primer caso reportado de infección por este patógeno en un perro. La infección se manejó con éxito médicamente, sin intervención quirúrgica.

Feohifomicose foi diagnosticada em um cão da raça Dachshund, macho castrado, de seis anos de idade, em tratamento imunossupressivo. O fungo identificado por cultura e PCR foi Phialophora americana. Este é o primeiro relato de caso de infecção por este patógeno em um cão. A infecção foi bem conduzida com tratamento medicamentoso, sem intervenção cirúrgica.

Venom immunotherapy for Hymenoptera allergy in a dog.

A 1.5-year-old male castrated dog was presented in anaphylactic shock after suffering an apparent bee sting. Immunotherapy with bee venom was initiated based upon history, skin testing and serological testing for allergen-specific immunoglobulin (Ig)E. The dog was maintained on venom immunotherapy for five years and showed no signs of adverse effects from therapy or from repeated bee stings.

Un chien castré de 1,5 ans a été présenté pour choc anaphylactique après avoir été piqué par une abeille. L'immunothérapie avec le venin d'abeille a été initié en fonction des commémoratifs, des tests cutanés et des tests sérologiques pour les immunoglobulines (Ig)E spécifiques d'allergènes. Le chien a été maintenu sous immunothérapie au venin pendant cinq ans et n'a montré aucun effet indésirable du traitement ou a la suite d'autres piqures d'abeilles.

Un perro macho castrado de 1,5 años se presentó en shock anafiláctico luego de sufrir una aparente picadura de abeja. La inmunoterapia con veneno de abeja se inició basándose en el historial, las pruebas cutáneas y las pruebas serológicas para la inmunoglobulina (Ig)E específica de alérgenos. El perro se mantuvo con inmunoterapia con veneno durante cinco años y no experimentó efectos adversos con la terapia o con repetidas picaduras de abeja.

Um cão macho castrado de 1 ano e meio de idade foi apresentado em choque anafilático após aparentemente ter sido picado por abelha. Iniciou-se a imunoterapia com veneno de abelha baseado na história clínica, testes alérgicos cutâneos e sorológicos para imunoglobulina (Ig)E alérgeno-específica. O cão foi mantido em imunoterapia com veneno por cinco anos e não apresentou nenhum efeito adverso do tratamento ou de novas picadas de abelha.

Sensitivity and specificity of a shortened elimination diet protocol for the diagnosis of food-induced atopic dermatitis (FIAD).

BACKGROUND: The gold standard to diagnose food allergy in dogs is an eight week elimination diet trial (EDT) followed by a re-challenge. A recent study demonstrated that a shorter EDT is possible if prednisolone is administered initially. HYPOTHESIS/OBJECTIVES: The goal was to evaluate the sensitivity and specificity of the EDT based on the number of relapses after prednisolone discontinuation. In addition, the aim was to determine whether the initial treatment length or the replacement of prednisolone with oclacitinib would influence the outcome. ANIMALS: Eighty-seven dogs with atopic dermatitis. METHODS AND MATERIALS: Dogs were fed an elimination diet and treated with either prednisolone or oclacitinib for two to three weeks. Relapsing dogs were treated a second time. In the absence of a relapse after two weeks off medication, dogs then were challenged. Dogs never achieving two weeks off treatment without relapse received the regular EDT. RESULTS: Fifty-eight of 87 dogs completed the study. Thirty-nine of 58 dogs received prednisolone; 21 of these were diagnosed with FIAD and had no relapse (n = 14), one relapse (n = 6) or two relapses (n = 1). Nineteen of 58 dogs received oclacitinib; 11 of these were deemed food-allergic and had no relapse (n = 7) or two relapses (n = 4). The initial treatment duration did not influence the outcome. The threshold of one relapse or less for the diagnosis of FIAD was associated with a sensitivity of 95% for prednisolone and 63% for oclacitinib. The specificity was 100% for both drugs. CONCLUSION AND CLINICAL IMPORTANCE: Initial prednisolone or oclacitinib use in EDT shortens the time to diagnosis of FIAD.

Atopic dermatitis in West Highland white terriers - Part III: early life peripheral blood regulatory T cells are reduced in atopic dermatitis.

BACKGROUND: Regulatory T (Treg) cells are involved in homeostasis of immune regulation and suppression of inflammation and T-cell polarisation. Current knowledge regarding the role of Treg cells in the initiation of allergic disease is limited for both people and dogs. OBJECTIVES: To explore the role of circulating Treg cells and their possible influencing factors, on the development of atopic dermatitis (AD). METHODS AND MATERIALS: This study followed part of a birth cohort of West Highland white terrier dogs and classified them according to eventual clinical signs of AD (i.e. allergic versus healthy). The Treg phenotypes were assessed longitudinally by flow cytometry at 3, 3-12 and 12-36 months of age, and associated with development of AD. Different early life antigenic factors [endotoxins and allergens in house dust, Toxocara canis-specific immunoglobulin (Ig)E/IgG, allergen-specific and total IgE, skin microbiota] were measured at three months of age, and a possible association with Treg cell levels was assessed. RESULTS: The percentages of CD4+ CD25+ Foxp3+ Treg cells in healthy dogs were significantly higher at in 3-month-old (mean 4.5% healthy versus 3.3% allergic; P = 0.021) and <1-year-old (4.0% healthy versus 2.9% allergic; P = 0.028) dogs when compared to percentages of Treg cells in dogs that developed AD. There was a significantly positive correlation between the relative abundance of Lachnospiraceae on the skin and CD4+ CD25+ Foxp3+ Treg cells in puppies that became allergic (r = 0.568, P = 0.017). CONCLUSION AND CLINICAL IMPORTANCE: Further large-scale studies are needed to identify the practical value of these findings in AD diagnosis, treatment and prevention.

A comparative study of subcutaneous, intralymphatic and sublingual immunotherapy for the long-term control of dogs with nonseasonal atopic dermatitis.

BACKGROUND: Allergen-specific immunotherapy (ASIT) is the only causative treatment of canine atopic dermatitis (cAD). Different routes for administration of ASIT have been used; however, comparative studies are lacking. HYPOTHESIS/OBJECTIVES: The present study compared the efficacy and safety of subcutaneous (SCIT), intralymphatic (ILIT) and sublingual (SLIT) immunotherapy. ANIMALS: 30 atopic dogs were included and allocation to three groups (SCIT, n = 8; ILIT, n = 12; SLIT, n = 10) was determined by the owners. METHODS AND MATERIALS: ASIT was administered using routine protocols. The pruritus Visual Analog Scale (PVAS), canine atopic dermatitis extent and severity index (CADESI), concurrent medications and adverse events were recorded initially and one, three, six and 12 months later. The main outcome measure was return to a normal status, which included CADESI <12, PVAS <2.5 and medication score <10. RESULTS: Drop-outs were distributed evenly and 23 dogs finished the study (SCIT, n = 6; ILIT, n = 10; SLIT, n = 7). Adverse reactions to treatment were rare. At the start of the study, the three groups were homogeneous with respect to clinical signs and concurrent medications. After 12 months of ASIT, the CADESI and PVAS had decreased with a stable medication score in the ILIT and SCIT groups (P < 0.05), while all three scores had increased in the SLIT group. Return to normal state was achieved in one of six (17%) dogs receiving SCIT, in six of 10 (60%) dogs receiving ILIT and in one of seven (14%) dogs receiving SLIT. CONCLUSIONS AND CLINICAL IMPORTANCE: These findings suggest that SCIT and ILIT improved clinical signs of cAD, whereas ILIT had a much higher return to normal rate.

Atopic dermatitis in West Highland white terriers - part I: natural history of atopic dermatitis in the first three years of life.

BACKGROUND: Canine atopic dermatitis (cAD) is a common allergic skin disease that is known to affect individuals early in life; the natural history of its initial development has not been documented. Some breeds such as West Highland white terriers (WHWTs) are highly predisposed to cAD. OBJECTIVES: To follow 100 WHWT puppies during their first three years and to record the onset of clinical signs of cAD. ANIMALS: One hundred and eight puppies from 29 litters were included and 90 were followed for three years. METHODS AND MATERIALS: Puppies were examined initially while with their breeders. After adoption, the owners were contacted twice each year and dogs were examined by veterinarians if signs compatible with cAD were detected; diagnosis of cAD was by two different definitions. The onset, location of the clinical signs and severity of cAD, as well as co-morbidities were recorded. RESULTS: The prevalence of cAD in the cohort was 52%. Most affected dogs (60%) developed signs of cAD during their first year of life and males were over-represented. The location of clinical signs mirrored those of previous descriptions. The severity of cAD was mild in 36% and severe in 13% of affected WHWTs. Dogs with cAD often exhibited other atopic diseases, but only gastro-intestinal signs were significantly different between WHWTs with and without cAD. Adverse reaction to foods was diagnosed in 24% of dogs. CONCLUSION AND CLINICAL IMPORTANCE: This longitudinal study of puppies from a predisposed breed sheds new light on the early development of cAD in WHWTs.

Atopic dermatitis in a cohort of West Highland white terriers in Switzerland. Part II: estimates of early life factors and heritability.

BACKGROUND: There is accumulating evidence in studies of allergic diseases in humans and dogs that environmental experiences during the first months of life can influence the development of allergic disease. No prospective study has evaluated this in veterinary medicine. HYPOTHESIS/OBJECTIVES: To assess early-life risk factors for canine atopic dermatitis (cAD) and estimate its heritability. ANIMALS: A West Highland white terrier birth cohort (n = 107) followed up to three years of age recording the development of cAD. METHODS AND MATERIALS: The effect of environmental factors [house dust mites (HDM), hygiene, feeding, lifestyle] and early-life determinants [breeder, mode of delivery, birth season, sex, litter size, early-life immunoglobulin E (IgE) levels] were assessed, using Stata SE 15.1 statistical analysis. Heritabilities were estimated using the R program packages MCMCglmm and QGglmm. RESULTS: Maternal allergic status [P = 0.013, odds ratio (OR 3.3)], male sex (P = 0.06), mode of delivery (P = 0.12), breeder (P = 0.06), presence of HDM (P = 0.11) and environmental hygiene level (P = 0.15) were identified as possible influence factors by bivariate analyses. In the multivariate analysis the male sex was significantly associated with the development of cAD in the offspring (P = 0.03, OR 2.4). The heritabilities on the observed scale were 0.31 (direct), 0.04 (maternal genetic effects) and 0.03 (maternal permanent environmental effects). CONCLUSION AND CLINICAL IMPORTANCE: These results suggest that several environmental factors could influence the development of cAD but clearly demonstrate the genetic influence of the individual and the dam. Further studies are needed to identify specific environmental factors, which could be potential targets for primary disease intervention.

RNA-seq analysis in equine papillomavirus type 2-positive carcinomas identifies affected pathways and potential cancer markers as well as viral gene expression and splicing events.

Equine papillomavirus type 2 (EcPV2) was discovered only recently, but it is found consistently in the context of genital squamous cell carcinomas (SCCs). Since neither cell cultures nor animal models exist, the characterization of this potential disease agent relies on the analysis of patient materials. To analyse the host and viral transcriptome in EcPV2-affected horses, genital tissue samples were collected from horses with EcPV2-positive lesions as well as from healthy EcPV2-negative horses. It was determined by RNA-seq analysis that there were 1957 differentially expressed (DE) host genes between the SCC and control samples. These genes were most abundantly related to DNA replication, cell cycle, extracellular matrix (ECM)-receptor interaction and focal adhesion. By comparison to other cancer studies, MMP1 and IL8 appeared to be potential marker genes for the development of SCCs. Analysis of the viral reads revealed the transcriptional activity of EcPV2 in all SCC samples. While few reads mapped to the structural viral genes, the majority of reads mapped to the non-structural early (E) genes, in particular to E6, E7 and E2/E4. Within these reads a distinct pattern of splicing events, which are essential for the expression of different genes in PV infections, was observed. Additionally, in one sample the integration of EcPV2 DNA into the host genome was detected by DNA-seq and confirmed by PCR. In conclusion, while host MMP1 and IL8 expression and the presence of EcPV2 may be useful markers in genital SCCs, further research on EcPV2-related pathomechanisms may focus on cell cycle-related genes, the viral genes E6, E7 and E2/E4, and integration events.

Paving the way for more precise diagnosis of EcPV2-associated equine penile lesions.

BACKGROUND: There is growing evidence that equine papillomavirus type 2 (EcPV2) infection is causally associated with the development of equine genital squamous cell carcinomas (SCCs). Early stages of disease present clinically as plaques or wart-like lesions which can gradually progress to tumoural lesions. Histologically these lesions are inconsistently described as benign hyperplasia, papilloma, penile intraepithelial neoplasia (PIN), carcinoma in situ (CIS) or SCC. Guidelines for histological classification of early SCC precursor lesions are not precisely defined, leading to potential misdiagnosis. The aim of this study was to identify histologic criteria and diagnostic markers allowing for a more accurate diagnosis of EcPV2-associated equine penile lesions. RESULTS: A total of 61 archived equine penile lesions were histologically re-assessed and classified as benign hyperplasia, papilloma, CIS or SCC. From these, 19 representative lesions and adjacent normal skin were comparatively analysed for the presence of EcPV2 DNA and transcripts using PCR and RNA in situ hybridisation (RISH). All lesional samples were positive by EcPV2 PCR and RISH, while adjacent normal skin was negative. RISH analysis yielded signal distribution patterns that allowed distinction of early (hyperplasia, papilloma) from late stage lesions (CIS, SCC). Subsequently, the 19 lesions were further assessed for expression of p53, Ki67, MCM7 and MMP1 by immunohistochemistry (IHC). All four proteins were expressed in both normal and lesional tissue. However, p53 expression was up-regulated in basal keratinocyte layers of papillomas, CIS and SCCs, as well as in upper keratinocyte layers of CIS and SCCs. MCM7 expression was only up-regulated in upper proliferating keratinocyte layers of papillomas, CIS and SCCs. CONCLUSION: This study proposes combining a refined histological protocol for analysis of equine penile lesions with PCR- and/or RISH based EcPV2-screening and p53/MCM7 IHC to more accurately determine the type of lesion. This may help to guide the choice of optimum treatment strategy, especially at early stages of disease.

The usefulness of short-course prednisolone during the initial phase of an elimination diet trial in dogs with food-induced atopic dermatitis.

BACKGROUND: Food allergy is a possible cause of atopic dermatitis (AD) in dogs; it is typically diagnosed following an eight-week elimination diet trial (EDT) and a provocation with the original diet. This lengthy procedure is difficult for owners and its interpretation may be unclear. HYPOTHESIS/OBJECTIVES: To test the effect of prednisolone used in the first weeks of an EDT in order to reduce the total time period for diagnosis. The goal was to perform food challenges earlier than after the traditionally recommended eight weeks. ANIMALS: Fifty-three dogs with AD were included in the study. METHODS AND MATERIALS: All dogs were fed a commercially available extensively hydrolyzed protein-based commercial pet food and treated with prednisolone for at least two weeks to control pruritus and inflammation. Dogs were challenged two weeks after prednisolone finished, provided that no flare had occurred. Dogs with relapsing signs were fed the hydrolyzate for at least eight weeks with or without further prednisolone treatment. RESULTS: Ten of 53 dogs (19%) had no relapse after two weeks off prednisolone: they were subsequently challenged with their regular food, had a relapse of signs and were diagnosed with a food-induced AD within four to six weeks of starting the EDT. In the other dogs, signs remained uncontrolled without prednisolone or relapsed rapidly after its discontinuation: they were considered nonfood-allergic after an eight week EDT. CONCLUSION AND CLINICAL IMPORTANCE: This study demonstrates that a shorter EDT is possible if the allergic pruritus and inflammation are initially controlled with a short course of glucocorticoids. This shortened trial is likely to improve owner adherence and facilitate the diagnosis of food allergy.

Glucocorticosteroids and ciclosporin do not significantly impact canine cutaneous microbiota.

BACKGROUND: As prednisone and ciclosporin can have immunosuppressive effects and have been considered potential predisposing factors for skin infections, we investigated the impact of these drugs on the diversity of the cutaneous microbiota, the abundance of Malassezia and infection with Papillomaviruses. RESULTS: Six atopic, asymptomatic Maltese-beagle dogs were treated with ciclosporin for one month and then with prednisone for another month, with a one-month wash-out between treatments. The dogs were sampled on the abdomen and pinna before and after each treatment using a swab. Samples for Papillomavirus detection were obtained with cytobrush sticks. The bacterial microbiota was characterized using 16S amplicon high-throughput sequencing. Malassezia populations were quantified with nested real-time PCR targeting the ribosomal internal transcribed spacer 1. The diversity and composition of cutaneous microbiota was not impacted in a detectable manner by any of the treatments. As observed for the bacterial microbiota, Malassezia populations were not affected by treatment. Three dogs were positive for Papillomavirus at more than one timepoint, but an association with treatment was not apparent. CONCLUSIONS: Ciclosporin and prednisone at doses used for the treatment of atopic dermatitis do not impact the canine cutaneous microbiota in a detectable manner.

Development of a core outcome set for therapeutic clinical trials enrolling dogs with atopic dermatitis (COSCAD'18).

BACKGROUND: For decades, the efficacy of interventions in clinical trials enrolling dogs with atopic dermatitis (AD) relied on heterogeneous evaluations of skin lesions and pruritus using unvalidated tools. Although some instruments for clinical signs were validated later, there was little impact on standardizing outcome measures resulting in difficulties in comparing treatment efficacy between trials and impeding meta-analyses. RESULTS: Participants in the Outcome Measures subcommittee of the International Committee of Allergic Diseases of Animals (ICADA) collaborated for two years to develop a core outcome set (COS) for canine AD, the COSCAD. This project involved several steps, constantly-re-assessed during online exchanges, to define the scope of this COS, to identify the relevant stakeholders, the domains to be evaluated, the instruments available for measuring agreed-upon domains and how to express outcome measures. This COSCAD'18 was designed principally for therapeutic-but not preventive or proactive-clinical trials enrolling dogs with chronic, nonseasonal (perennial), moderate-to-severe AD. Selected domains were skin lesions, pruritus manifestations and perception of treatment efficacy. Instruments to evaluate these domains were the CADESI4 or CADLI, the 10-point pruritus visual analog scale (PVAS10) and the Owner Global Assessment of Treatment Efficacy (OGATE), respectively. The COSCAD'18 has three outcome measures: the percentages of dogs with veterinarian-assessed skin lesions or owner-rated pruritus manifestation scores in the range of normal dogs or those with mild AD; the third is a good-to-excellent global assessment by the pet owners of their perception of treatment efficacy. Importantly, this COSCAD'18 is not meant to represent the sole-or primary-outcome measures evaluated in a trial; authors are always free to add any others, which they deem will best assess the efficacy of tested interventions. Benchmarks to define a threshold for treatment success were not set, as what constitutes a clinically-relevant therapeutic efficacy is expected to vary greatly depending interventions. CONCLUSIONS: This COSCAD'18 should help veterinarians and owners compare the benefits of treatments in future trials. This COS should also facilitate the combination of trial results in future systematic reviews, thereby producing more reliable summary estimates of treatment effects and enhancing evidence-based veterinary dermatology.

An open study on the efficacy of a recombinant Der f 2 (Dermatophagoides farinae) immunotherapy in atopic dogs in Hungary and Switzerland.

BACKGROUND: Previously published studies evaluating a recombinant Der f 2-based immunotherapy have demonstrated efficacy in the treatment of dogs experimentally and naturally sensitized to house dust mites (HDM). Der f 2 sensitization is thought to play a minor role in European atopic dogs sensitized to HDM. OBJECTIVE: The study evaluated the short-term efficacy of a recombinant Der f 2 product in the treatment of naturally sensitized atopic dogs in Switzerland and Hungary. ANIMALS: Fifteen atopic dogs with positive test reactions to Dermatophagoides farinae (Df). MATERIAL AND METHODS: Recombinant Der f 2 allergens were injected subcutaneously at increasing doses once weekly for 6 weeks. Canine Atopic Dermatitis Extent and Severity Index (CADESI-04), pruritus Visual Analog Scale (pVAS) and medication scores were assessed at days 0 and 42. Efficacy was recorded as excellent, good, fair or poor, depending on the number of scores decreasing by more than 50%. RESULTS: Mean CADESI, pVAS and medication scores at inclusion were 35, 6 and 15 (SD = 30, 2, 7), respectively. At Day 42 the scores decreased to 8, 3 and 5, respectively (Wilcoxon matched pairs signed rank tests P = 0.0002, 0.002 and 0.001). Four dogs were classified as excellent responders with a reduction of >50% in all three scores. Nine dogs were classified as good (five) or fair (four) responders and scores deteriorated in two dogs. CONCLUSION: These data suggest that recombinant Der f2 allergens may be as effective and show benefit faster than traditional allergen immunotherapy in European dogs sensitized to Df.

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-ß 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-ß 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.

Total and Toxocara canis larval excretory/secretory antigen- and allergen-specific IgE in atopic and non-atopic dogs.

BACKGROUND: Total IgE concentrations are higher in dogs than in humans. Persistent Toxocara canis larval infection is prevalent in dogs and is associated with substantial specific antibody reactions. A correlation, however, between total IgE and T. canis-specific antibody levels in dogs has not been evaluated. OBJECTIVES: To determine the relationship between total IgE, T. canis-specific IgG and IgE, and allergen-specific IgE levels in atopic and non-atopic dogs, and to evaluate possible confounding factors. ANIMALS: Sera of 30 atopic and 30 non-atopic client-owned dogs. METHODS: Total IgE, T. canis-specific antibody and allergen-specific IgE levels were evaluated by ELISA. RESULTS: Total IgE, T. canis-specific antibody and allergen-specific IgE levels were significantly higher in non-atopic compared to atopic dogs. A positive correlation was demonstrated between T. canis-specific IgG and T. canis-specific IgE; T. canis-specific IgG and total IgE; T. canis-specific IgE and total IgE; and allergen-specific IgE and total IgE. No differences were detected on the basis of age, gender, vaccination status; deworming or season between atopic and non-atopic dogs. Previous immunomodulatory treatment and cause of atopy did not influence antibody levels of atopic dogs. CONCLUSIONS: Toxocara canis-specific IgE appears to be a major component of total IgE in dogs. Total and T. canis-specific IgE levels are higher in non-atopic compared to atopic dogs. It is speculated that T. canis infection may have a protective effect against the development of canine atopic dermatitis and/or that elevations in total serum IgE level are often not associated with atopic dermatitis.