Are transepidermal water loss and clinical signs correlated in canine atopic dermatitis? A compilation of studies

Background – Cutaneous impairment plays a crucial role in atopic dermatitis (AD). Transepidermal water loss (TEWL) measurement is an indirect assessment of skin barrier function and correlates with disease severity in humans. Skin impairment also exists in canine AD; however, concerns exist regarding variability and reliability of TEWL measurements in dogs. Hypothesis/Objectives – The purposes of this retrospective study were twofold: first, to investigate the correlation between severity of dermatitis [measured by Canine Atopic Dermatitis Extent and Severity Index (CADESI)] and TEWL; and second, to evaluate whether increased TEWL at a young age correlates with disease severity later in life. Methods – Data from atopic beagles and dogs with natural AD were analysed. Transepidermal water loss was measured in atopic beagles (n = 24) with an open chamber and in dogs with naturally occurring AD with a closed chamber device (two studies, with n = 14 and n = 18). Pearson product–moment correlation was used for analyses. Transepidermal water loss of the inguinal region, axilla, antebrachial flexure and pinna was analysed. Correlations were investigated for each study, separately first and then jointly. They included CADESI and TEWL of individual regions, total CADESI and total TEWL of all measured regions, and total CADESI and TEWL of key regions. Results – In atopic beagles, TEWL measured at 1 year of age pre‐ and post‐allergen challenge was correlated with CADESI at 1, 3 and 6 years of age. Overall, low correlation coefficients were found; therefore, a biologically relevant connection could not be demonstrated. The main significant positive correlation was found between TEWL in the pinna and total CADESI. Conclusions and clinical importance – It is concluded that TEWL does not correlate with disease severity.     

Environmental and oral challenge with storage mites in beagles experimentally sensitized to Dermatophagoides farinae

This study aimed to investigate whether challenge with storage mites elicited flare ups of atopic dermatitis (AD) in Dermatophagoides farinae sensitized atopic Beagles housed in a house dust and storage mite-free environment. Atopic Beagles were environmentally challenged with 50 mg of Tyrophagus putrescentiae for three days in a row. Clinical signs were scored before, 6 h after each challenge and then every 24 h for a total of 5 days using a Canine Atopic Dermatitis Extent and Severity Index. Four healthy Beagles, negative on serology and intradermal testing for both house dust and storage mites, were used as controls and similarly challenged. A month after environmental challenge, the atopic Beagles were challenged by the oral route (50 mg of T. putrescentiae for three days in a row) and evaluated as described. Analyses of variance (ANOVAs) were used for comparisons between groups and types of challenges. All atopic Beagles developed erythematous pruritic lesions clinically compatible with AD on the face, pinnae, feet and ventral abdomen after both environmental and oral challenge. Control dogs did not develop dermatitis except for mild pinnal erythema in one dog. In the environmental challenge, ANOVA showed a significant effect of time, group, and group x time interaction, with atopic Beagles showing significantly higher scores than the controls. There were no significant differences in clinical scores after oral and environmental challenge in the atopic group. Cross-reactivity between house dust and storage mites could therefore contribute to flare ups of AD in house dust mite allergic dogs.     

Peripheral blood mononuclear cell responses to Dermatophagoides farinae in canine atopic dermatitis

Atopic dermatitis is a chronic inflammatory and pruritic skin disease commonly seen in dogs and humans that is characterised by the presence of allergen-specific IgE. Data from skin tests and serological analysis suggest that the house dust mite Dermatophagoides farinae is the most important allergen in dogs with atopic dermatitis. The aim of this study was to determine if D. farinae specific peripheral blood mononuclear cell (PBMC) responses could be detected in dogs with atopic dermatitis. PBMCs were isolated by the density centrifugation from dogs with atopic dermatitis that were skin test positive for D. farinae, dogs with atopic dermatitis that were skin test negative for D. farinae, and healthy dogs. Cells were cultured with increasing concentrations of the D. farinae extract, no antigen, vaccine antigens or concanavalin A (ConA). There was significantly greater responsiveness of PBMCs from the D. farinae positive dogs than from either the D. farinae negative or healthy dogs (ANOVA, P<0.05). In contrast, no significant differences were observed in the control responses between the three groups. This is the first study to demonstrate that D. farinae specific circulating memory cells are involved in the pathogenesis of canine house dust mite hypersensitivity.     

Transepidermal water loss in healthy and atopic dogs, treated and untreated: a comparative preliminary study

An impaired skin barrier function is thought to be crucial for allergic sensitization. In humans, the skin barrier is assessed by noninvasive methods, such as the measurement of transepidermal water loss (TEWL). Although limitations have been reported, measurement of TEWL has been demonstrated to be a suitable method to assess barrier function indirectly in dogs. The purposes of this prospective clinical study were twofold. The first aim was to evaluate and compare TEWL in healthy and atopic dogs. The second aim was to evaluate TEWL in a population of atopic dogs and to assess TEWL in dogs whose disease was in remission after successful therapy and compare it with dogs whose disease was not controlled or receiving treatment. One hundred and fifty dogs were selected and divided into the following three groups: 50 atopic dogs before specific treatment (group A); 50 in remission (group B); and 50 control dogs (group C). The mean values for TEWL for each group were 22.47 (g/m(2) h) (group A; 95% confidence interval 20.85-24.09), 12.57 (g/m(2) h) (group B; 95% confidence interval 11.43-13.7) and 8.81 (g/m(2) h) (group C; 95% confidence interval 8.09-9.52); P-value was 0.0001 for TEWL (groups A, B and C). This study showed a significant difference of TEWL between healthy control dogs and dogs with atopic dermatitis. Additionally, TEWL was lower in atopic dogs whose disease was in remission due to treatment. These results were consistent with reports in human medicine about TEWL.     

Sulphido-leukotriene production from peripheral leukocytes and skin in clinically normal dogs and house dust mite positive atopic dogs

Pathogenesis of canine atopy has not been completely elucidated. In humans, sulphido-leukotrienes (s-LT) play a role in atopy, and increased production of s-LT occurs in the skin and peripheral leukocytes after allergen challenge. The study population included 16 clinically normal and 13 atopic dogs. All atopic dogs had in common a positive reaction (4+) to the intradermal injection of house dust mite (allergen of reference). Blood samples and skin biopsies were collected. Sulphido-LT synthesis by peripheral leukocytes after stimulation was measured, and no statistically significant difference was found between clinically normal and atopic dogs. Sulphido-LT concentrations in skin samples from stimulated and unstimulated sites were measured, and no statistically significant difference was detected between clinically normal and atopic dogs or between lesional and nonlesional skin within the atopic group. Clinical signs of atopic dogs were graded by owners and no correlation was found between their severity and cutaneous concentrations of s-LT. In this study there was no increase in s-LT synthesis in atopic dogs.     

Transmission electron microscopy studies in an experimental model of canine atopic dermatitis

Impairment of skin barrier function has been hypothesized in canine atopic dermatitis (AD). In this prospective, controlled study, the ultrastructure of the upper epidermal layers was investigated using an experimental model of canine AD. Seven atopic Beagles sensitized to Dermatophagoides farinae and four healthy Beagles were used as controls. Both normal and atopic dogs were challenged with D. farinae for 3 days. Clinical signs were scored and skin biopsies were taken from the inguinal area before and 3 days after allergen exposure. Samples were processed to enhance lipid visibility and evaluated by Transmission Electron Microscopy. Emphasis was placed on evaluation of the lipid lamellae (LL), and lamellar bodies (LB) of the stratum corneum.
After allergen challenge, atopic Beagles developed severe pruritic dermatitis while no skin lesions were noted in the controls. Ultrastructurally, before allergen challenge, atopic Beagles displayed focally severe abnormalities in LL organization and wider intercellular spaces containing abnormal lipid material. In atopic Beagles, LBs were frequently found inside corneocytes while this finding was not observed in the controls. After allergen challenge, further increase of intercellular spaces was observed in the stratum corneum of atopic Beagles while no appreciable changes were observed in the normal dogs. Intercellular spaces in atopic Beagles were filled with abundant amounts of abnormal lipid material and highly disorganized LL. It is concluded that baseline differences in the ultrastructure of the skin exist between normal and experimentally sensitized atopic Beagles and that these changes are aggravated by allergen challenge and the resulting flare-up of dermatitis.     

Use of an amplified ELISA technique for detection of a house dust mite allergen (Der f 1) in skin and coat dust samples from dogs

OBJECTIVE: To use an amplified ELISA technique to document the presence and quantify the concentration of the house dust mite allergen, Der f 1, in skin and coat dust samples collected from dogs. ANIMALS: 29 pet dogs of various breeds. PROCEDURE: Dogs were weighed, and body surface area in square meters was determined. Skin and coat dust samples were obtained by vacuuming dogs. Collected dust was analyzed by use of standard and amplified ELISA techniques. RESULTS: By use of the standard ELISA technique, Der f 1 was detected in skin and coat dust samples from 6 of 29 (21%) dogs. Mean concentration of Der f 1 in the 6 samples with positive assay results was 16.16 ng/mL (range, 5.61 to 31.24 ng/mL). Samples with negative assay results were retested for dust mite allergen by use of an amplified ELISA technique; an additional 14 dogs had positive assay results. Mean concentration of allergen was 0.36 ng/mL (range, 0.19 to 2.20 ng/mL). Combining both techniques, 20 of 29 (69%) dogs had positive assay results for Der f 1. CONCLUSIONS AND CLINICAL RELEVANCE: Results of our study indicate that house dust mite allergens are present on the skin and in the coat of dogs, and this source of allergen may act as a reservoir for allergen exposure in hypersensitive dogs. Use of an amplified ELISA technique to determine environmental concentrations of house dust mite allergens in homes and on dogs will help to identify the relationship between immunologic findings and environmental exposures in dogs with atopic dermatitis.     

Evaluation of a hand‐held evaporimeter (VapoMeter®) for the measurement of transepidermal water loss in healthy dogs

In humans, transepidermal water loss (TEWL) is measured by noninvasive techniques using either open‐ or closed‐chamber instruments. The aim of this study was to investigate the use of a hand‐held, closed chamber device (Vapometer^®) to measure TEWL in canine skin. Repeated measurements obtained from multiple body sites in one short and one long‐coated dog had mean coefficients of variation ranging from 20% to 33%. In the short‐coated dog, TEWL ranged from a mean of 5.8 g/m²/h on the ventral abdomen to 24.4 g/m²/h between the shoulders. In the long‐coated dog, mean TEWL values ranged from 26.3 g/m²/h on the right chest wall to 51.3 g/m²/h in the right axilla. TEWL readings differed significantly at different body sites and showed significant day‐to‐day variation. In a comparison of a further 20 dogs, TEWL readings obtained from the lateral thorax differed significantly between dogs. Furthermore, in seven of the twenty dogs, readings differed significantly when one side was compared with the other. The Vapometer^® was able to measure TEWL in canine skin and yielded values similar to those previously reported in the literature using other devices. However, for use in clinical studies, the significant site to site, day‐to‐day and dog to dog variations would make changes induced by disease, drugs, dietary supplements or topical agents very difficult to reliably detect.     

Dermatophagoides farinae house dust mite allergen challenges reduce stratum corneum ceramides in an experimental dog model of acute atopic dermatitis

Background – Ceramides are essential stratum corneum (SC) lipids and they play a pivotal role in maintaining effective cutaneous barrier function. Objectives – The present study aimed at determining the effect of a Dermatophagoides farinae house dust mite (Df‐HDM) allergen challenge on SC ceramides of atopic dogs experimentally sensitized to these allergens. Animals – Six Df‐HDM‐sensitized atopic Maltese–beagle dogs were used. Methods – Prechallenge SC was obtained by cyanoacrylate stripping. One week later, the dogs were challenged topically with Df‐HDM allergens, which resulted in mild to moderate inflammation 24 h later. Two weeks after challenge, SC of lesional and nonlesional skin was obtained. Finally, SC was collected from challenge sites 2 months after lesion resolution. The different SC lipids were quantified blindly by thin‐layer chromatography. Results – Significantly lower amounts of ceramides [AH], [AP], [AS], [NP], [EOP], [NS] and [EOS] were observed in lesional SC compared with prechallenge samples, while no significant effect was found on the amount of other lipids, including cholesterol and free fatty acids. The ceramide profile of nonlesional skin generally showed the same postchallenge reduction pattern. Ceramide amounts returned to normal within 2 months after lesion remission. Conclusion and clinical importance – These findings suggest that the allergic reactions caused by Df‐HDM allergens lead to a selective reduction of SC ceramides, not only at sites of inflammation but also at sites away from those of allergen application. There is normalization of ceramide amounts after inflammation subsides. These observations suggest that the deficiency of ceramides observed in canine atopic skin occurs, at least in part, secondary to inflammation.     

Use of contact plates for the quantitative culture of Malassezia pachydermatis from canine skin

The Malassezia pachydermatis populations of the axilla and groin of 12 normal and 12 atopic dogs were compared using tape-strips and contact plates. When assessed by either method, the mean density of yeasts in the groin of the atopic dogs was significantly greater (P<0.05) than that of the normal dogs, suggesting that the cutaneous microenvironment of the groin region of the atopic dogs favoured colonisation by this yeast. Differences between the counts from the axilla were not significant. The frequency of isolation of yeasts from both dogs and sites was significantly higher (P<0.05 and P<0.001, respectively) in the atopic group. There was a very highly significant correlation (P<0.001) between the tape-strip counts and contact plate counts in the atopic group only. This study suggests that isolation of numerous M pachydermatis colonies from the axilla and groin of dogs using contact plates is indicative of elevated skin surface populations. The simplicity of the contact plate method makes it suitable for the routine quantitative culture of cutaneous M pachydermatis populations in dogs with dermatological disease.     

Validation of a novel epicutaneous delivery system for patch testing of house dust mite‐hypersensitive dogs

Background – Patch tests with allergens are used for the evaluation of cellular hypersensitivity to food and environmental allergens in dogs and humans with atopic dermatitis. Viaskin is a novel allergen epicutaneous delivery system that enhances epidermal allergen capture by immune cells. Objectives – To compare the use of Viaskin and Finn chamber patch tests in dogs hypersensitive to mite allergens. Methods – Empty control or Dermatophagoides farinae house dust mite‐containing Viaskin or Finn chamber patches were applied to the thoracic skin of six mite‐hypersensitive Maltese‐beagle crossbred atopic dogs. Lesions were graded 49 and 72 h after patch test application, and skin biopsies were collected after 72 h. Overall microscopic inflammation, eosinophil and T‐lymphocyte infiltrations were scored. Results – Positive macroscopic patch test reactions developed at five of six Viaskin application sites and four of six Finn chamber application sites. Median microscopic epidermal and dermal inflammation, as well as eosinophil and CD3 T‐lymphocyte dermal scores were always higher in biopsies collected at Viaskin than at Finn chamber sites. Microscopic inflammation scores were significantly higher after mite allergen‐containing Viaskin compared with empty patches, but this was not the case for mite‐containing Finn chambers compared with control chambers. Scores obtained using Viaskin were not significantly different from those obtained using Finn chambers. Macroscopic and microscopic scores were significantly correlated. Conclusions and clinical importance – In mite‐allergic dogs, Viaskin epicutaneous delivery systems appear to induce stronger allergen‐specific inflammation than currently used Finn chamber patch tests. Consequently, Viaskin patches might offer a better alternative for screening cellular hypersensitivity to food and environmental allergens.     

The influence of topical unsaturated fatty acids and essential oils on normal and atopic dogs

Seven dogs with atopic dermatitis and six normal dogs were treated with a spot-on product containing essential oils and unsaturated fatty acids q 7 days for 8 wk. Seven additional atopic dogs received a daily spray containing similar ingredients to the spot-on. In all dogs, transepidermal water loss (TEWL) was measured before and after treatment using a closed chamber device. In atopic dogs, a validated lesion score (canine atopic dermatitis extent and severity index, CADESI) was determined and pruritus was assessed with a visual analog scale before and after treatment. The mean CADESI scores in atopic dogs decreased with the spot-on (P=0.0043) and with the spray (P=0.0366). Similarly, the pruritus scores decreased with the spot-on (P=0.266) and with the spray (P=0.0177). There was a significant difference between the TEWL values of healthy and atopic dogs on the abdomen (P=0.0181) and back (P=0.0123). TEWL decreased significantly on the back after treatment with the spray (P=0.016), but not on the abdomen (P=0.078). Adverse effects were not observed. The results of this pilot study indicate that topical fatty acids and essential oils are a useful treatment option for canine atopic dermatitis.     

Studies on the role of routes of allergen exposure in high IgE-producing beagle dogs sensitized to house dust mites

The current study aimed to investigate the role played by oral, epicutaneous, and inhalation routes of exposure to house dust mites (HDM). The colony of high IgE-producing beagle dogs has been shown to develop pruritic dermatitis compatible with atopic dermatitis following environmental exposure (EE) to HDM. In crossover experiments, the response to EE was compared to two modified challenges, oral exposure (OE) and snood and muzzle exposure (SME). For OE, HDM were fed daily for 3 days. For SME, ingestion of allergen was prevented but there was inhalation and epicutaneous exposure to all body regions except to one ear. In all experiments, dogs were challenged for three consecutive days, and evaluated before, 6 h after exposure and daily thereafter, for 5 days. After a wash-out period, groups were crossed-over so that each dog was randomly challenged to all three protocols. Clinical scores were analysed using least squares analysis of variance. All dogs developed pruritic dermatitis regardless of the protocol. With OE, lesions developed in the same body regions as with EE although scores were lower. This difference became more evident after the first 3 days when OE scores decreased and EE scores continued to increase. The scores of covered and uncovered ears did not differ with SME. Scores for the remainder of the body were significantly lower than for EE. The development of lesions on covered ears supports the importance of inhalation or a systemic reaction to epicutaneous exposure in other areas. It is concluded that all routes are important and have additive effects, that route of exposure does not determine the distribution of lesions and that continuous epicutaneous exposure probably plays the most important role.     

Dermatophagoides mites in house dust as an allergen source in atopic dogs

Thirty dogs with a clinical diagnosis of atopy were skin tested with 58 allergens including an aqueous house dust mite extract and a crude house dust extract. Sixty percent of the dogs had positive skin reactions to both the house dust mite and house dust antigens. In atopic dogs, house dust mites appear to be an important allergen source in house dust extracts but are not the only major source.     

Common allergens of atopic dermatitis in dogs: comparative findings based on intradermal tests

Intradermal tests were performed on 58 dogs diagnosed with atopic dermatitis from 2004~2008 at the Veterinary Medical Teaching Hospital of Konkuk University, Korea. To compare the allergen distribution observed in the present investigation to the results from other studies conducted in Korea and elsewhere, the allergens were grouped according to their kinds. There was no significant difference in gender distribution among the dogs. The most common breeds among the 58 dogs were Maltese (n = 11) and Shih-tzu (n = 11). The average age was 4.8 years. The most frequently produced a positive reaction on the intradermal tests was mold (67.3%) followed by house dust (54.5%) and house dust mites (49.1%). The present study found a low distribution of dogs allergic to various outdoor allergens compared to studies performed in other countries; this may reflect differences in living conditions for dogs living in Korea.     

Allergen-induced production of IL-31 by canine Th2 cells and identification of immune,skin, and neuronal target cells

The canine cytokine IL-31 induces pruritus in dogs and can be detected in dogs with atopic dermatitis; however very little is understood around its interactions with specific canine cells. We hypothesize that IL-31 is involved in the progression of allergic skin disease by coordinating the interaction between the immune system with skin and neuronal systems. The goal of the following work was to identify cells that produce IL-31 as well as cells that may respond to this cytokine. Peripheral blood mononuclear cells (PBMCs) were collected from naïve and house dust mite (HDM) allergen-sensitized beagle dogs and used for ex vivo characterization of cytokine production assessed using ELISpot and quantitative immunoassay. Sensitization to HDM allergen induced a T-helper type 2 (Th2) cell phenotype characterized by an increase in the production of IL-4 protein. Interestingly, repeated allergen challenge over time also resulted in an increase in IFN-γ. Further evaluation showed that co-stimulation of Th2 polarized cells with antigen and the bacterial component Staphylococcus enterotoxin B (SEB) produced higher levels of IL-31 compared to either stimulant alone. Production of IL-31 when PBMCs were stimulated by T cell mitogens suggests T cells as a source of IL-31. Quantitative real-time PCR was utilized to determine expression of the IL-31 receptor alpha chain in canine cell lines and tissue. Canine monocytic cells, keratinocytes, and dorsal root ganglia were shown to express the IL-31 receptor alpha chain mRNA. In a multifaceted disease such as canine atopic dermatitis, the combination of Th2 polarization and microbial presence may lead to IL-31 mediated effects driving inflammation and pruritus by immune cells, keratinocytes, and direct neuronal stimulation.     

High allergen-specific serum immunoglobulin E levels in nonatopic West Highland white terriers

Human and canine atopic dermatitis (AD) share an association with IgE specific to environmental allergens, but few studies have evaluated serum allergen‐specific IgE in nonatopic dogs. This study compared serum allergen‐specific IgE levels in 30 atopic and 18 nonatopic West Highland white terriers. Atopic dermatitis was confirmed using standard criteria. Nonatopic dogs were over 5 years of age and had no clinical signs or history of AD. Serum allergen‐specific IgE levels were measured with Allercept^® IgE ELISAs using a 48‐allergen Australian panel. Positive reactions were defined as ≥150 ELISA absorbance units. Intradermal tests were performed in 16 atopic dogs, either at the time of or at various times prior to serum collection. In atopic dogs, the most common positive ELISA and intradermal test results were to Dermatophagoides farinae (11 of 30 dogs), but there were no statistically significant correlations between results from the two methods for any allergen. In nonatopic dogs, multiple high‐positive ELISA reactions were reported to 45 of 48 allergens, most commonly D. farinae and Tyrophagus putrescentiae (17 of 18 dogs each). Positive ELISA results in nonatopic dogs were statistically significantly higher than those in atopic dogs for 44 of 48 allergens, including two allergens (D. farinae and Dermatophagoides pteronyssinus) commonly regarded as significant in canine AD. In conclusion, positive allergen‐specific IgE ELISAs were not specific for canine AD, and high allergen‐specific IgE levels were seen in nonatopic dogs. The clinical significance of this and whether it characterizes a protective phenotype is unclear.     

Early exposure to probiotics in a canine model of atopic dermatitis has long-term clinical and immunological effects

Probiotics modulate the immune response and may have protective effects against atopic dermatitis (AD). Clinical trials using dogs with spontaneous disease are limited by confounding factors such as different diets, environments and sensitizations while a more controlled evaluation is possible using experimental models. A validated model of canine AD showed that early exposure to Lactobacillus rhamnosus GG (LGG) significantly decreases allergen-specific IgE and partially prevents AD in the first 6 months of life. This study is a follow-up three years after discontinuation of LGG. Clinical signs were evaluated after allergen challenge with ragweed, timothy, Dermatophagoides farinae. Allergen-specific IgE, IL-10 and TGF-β were measured on the 1st day of challenge, before allergen exposure. Normal dogs were included as controls. Analyses included seven dogs in the non-probiotic and nine in the probiotic litter. For clinical scores, a 2-Group × 9-Time Analysis of Variance showed significant effects of group (p=0.0003, probiotic相似文献   

Comparison of response to immunotherapy by intradermal skin test and antigen-specific IgE in canine atopy

The intradermal skin test (IDST) and serologic allergy test (SAT) has been developed for confirming a diagnosis of canine atopy and determining allergens for immunotherapy. To determine the prevalence of causative allergens for canine atopic dermatitis in Japan, IDST and SAT were performed with the CMG Immunodot strips on 95 atopic dogs using 9 allergens. In addition, we compared agreement rate, sensitivity and specificity between them (using IDST as the standard). The allergen most commonly positive in both tests was house dust mites (IDST: 69.5%, SAT: 48.4%). Moreover, Japanese cedar, mugwort and grass mix were detected as attendant causative allergens. Agreement rates between the two tests ranged from 67.4% to 96.8%; the overall mean agreement rate were 81%. SAT was shown to have sensitivity to IDST ranging from 16.7 to 68.2%. The specificities were very high for all allergens, on the order of 94.9-100% (median=98.7%). Finally, the efficacy of immunotherapy was evaluated on 27 atopic dogs based on IDST (15 dogs) and SAT (12 dogs) results. Overall, 60% (9/15) of the IDST group and 66.8% (8/12) of the SAT group experienced a 50% to 100% reduction in their symptomatology. No significant differences were found in response to immunotherapy during the follow-up period between allergen selection methods. These results indicate the value of serologic tests as an aid to identifying an allergen solution for immunotherapy.