Reactivity to intradermal injection of extracts of Dermatophagoides farinae, Dermatophagoides pteronyssinus, house dust mite mix, and house dust in dogs suspected to have atopic dermatitis: 115 cases (1996-1998)

OBJECTIVE: To compare reactivities to intradermal injection of extracts of Dermatophagoides farinae, Dermatophagoides pteronyssinus, house dust mite mix, and house dust in dogs suspected to have atopic dermatitis. DESIGN: Retrospective study. ANIMALS: 115 dogs. PROCEDURES: Records of all dogs suspected to have atopic dermatitis that underwent intradermal testing between October 1996 and July 1998 were reviewed. Reactivities to intradermal injection of crude mixed house dust mite (1:25,000 wt/vol) and crude house dust (25 PNU/ml) extracts were compared with reactivities to intradermal injection of individual extracts of D farinae and D pteronyssinus (1:50,000 wt/vol). RESULTS: Ninety dogs were confirmed to have atopic dermatitis including 61 of the 69 dogs with positive reactions to either or both of the individual house dust mite extracts. Intradermal testing with the mixed house dust mite extract had sensitivity of 75%, specificity of 96%, and accuracy of 83%. Intradermal testing with the house dust extract had sensitivity of 30%, specificity of 93%, and accuracy of 56%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that use of crude mixed house dust mite and crude house dust extracts for intradermal testing in dogs is not as accurate a method of determining house dust mite hypersensitivity as is the use of individual D farinae and D pteronyssinus extracts mainly because of the high percentage of false-negative results. Extracts of individual house dust mites are recommended for intradermal testing of dogs suspected to have atopic dermatitis.     

Patch test reactions to house dust mites in dogs with atopic dermatitis

The purpose of this study was to determine the percentage of dogs with spontaneous atopic dermatitis that show a positive patch test reaction to a commercially available 20% house dust mites mixture containing equal parts of Dermatophagoides farinae and Dermatophagoides pteronyssinus in white petrolatum. In addition, we evaluated whether skin reactions induced after the epicutaneous application of house dust mites were clinically and histologically similar to naturally developed skin lesions of dogs with atopic dermatitis. Furthermore, we investigated if the reactions induced by house dust mites were true allergic reactions by comparing them to atopic lesional skin and to patch test reactions induced by an irritant substance (sodium lauryl sulphate). White petrolatum alone and nonlesional skin sites were used as negative controls. Macroscopic and microscopic evaluations of the patch test and control sites were performed in a blinded fashion at 48 and 72 h after patch test application. Microscopic results were evaluated in a qualitative and quantitative manner. A chi‐square test for homogenicity was used for the quantitative analysis to compare the proportion of each dermal inflammatory cell type among positive histopathological tested sites. P values ≤ 0.05 were considered significant. The study included 12 healthy nonatopic dogs and 13 dogs with nonseasonal atopic dermatitis. None of the nonatopic dogs reacted to house dust mites and white petrolatum. Ten (77%) of the 13 atopic dogs reacted macroscopically and histopathologically to house dust mites. Macroscopic reactions induced by house dust mites were characterized by erythema, oedema and papules. The macroscopic reactions induced by house dust mites were identical to lesional skin in 20% of the dogs and identical to reactions induced by sodium lauryl sulphate in 40% of the dogs. Qualitative histopathological findings showed that the reactions induced by house dust mites were similar to atopic lesional skin in 80% of the dogs and were similar to sodium lauryl sulphate in 20% of the dogs. Quantitative analyses showed that the proportion of neutrophils in reactions induced by sodium lauryl sulphate was significantly higher (P < 0.05) compared to house dust mites reactions, which could be a differentiator factor between an allergic and an irritant reaction. These results showed that the epicutaneous application of house dust mites in dogs with atopic dermatitis induced histopathological lesions similar to spontaneous atopic lesions in dogs. Therefore, this study demonstrated that house dust mites penetrated the skin of dogs with atopic dermatitis and induced an inflammatory response that resembled a true allergic reaction. Funding: Small Companion Animal Grant, University of Minnesota.     

IgG responses to antigens from Dermatophagoides farinae in healthy and atopic dogs

In this study, we used a semi-quantitative electrophoresis and immunoblotting technique to characterise the IgG response to antigens from Dermatophagoides farinae in 20 healthy and 20 atopic dogs. Both groups mounted an IgG response to multiple antigens from the mite. There was no significant difference in the number of bands recognised, or the molecular weights of the bands, between the two groups. The two most obvious bands in both groups were proteins with molecular weights of 98 kDa (likely to be the high molecular weight allergen Der f 15) and 44 kDa, although dogs in both groups recognised a similar pattern of other antigens. The magnitude of the IgG response was greater in the atopic group although this was not statistically significant. The results indicate that the immune system of both healthy and atopic dogs generates an IgG response to multiple antigens from D. farinae. As some of these antigens (such as the 98 and 44 kDa proteins) are also targeted by IgE in atopic dogs, immunoglobulin class switching in response to Th2 cytokines may not be as dominant a process as has been proposed.     

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.     

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.     

Evaluation of IgG subclass responses against Dermatophagoides farinae allergens in healthy and atopic dogs

A semiquantitative chemiluminescent Western blot analysis system was developed and validated to evaluate antigen-specific IgG subclass responses to electrophoretically separated proteins of Dermatophagoides farinae in healthy and atopic dogs. Both groups mounted similar D. farinae-specific IgG1 and IgG4 responses to multiple antigens, but IgG2 and IgG3 responses were difficult to detect. The most commonly recognized bands in both groups were 18 and 98 kDa antigens for IgG1 and 18, 45, 66, 98, 130 and 180 kDa for IgG4. The number of bands recognized per dog did not differ significantly, but significantly more atopic dogs had an IgG1 response to a 180 kDa protein. The overall D. farinae-specific IgG1 and IgG4 responses were slightly higher, but not significantly different, in the healthy group. The results suggest that some antigens produced by D. farinae can induce different subclass responses. However, as most of these responses are seen in both healthy and atopic dogs, they are likely to merely represent recognition of foreign proteins presented to the immune system, rather than involvement in the pathogenesis of atopic dermatitis. The role of the 180 kDa antigen warrants further study.     

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.     

Effects of propofol-induced sedation on intradermal test reactions in dogs with atopic dermatitis

We compared the effect of propofol and saline control on intradermal test reactions in dogs with atopic dermatitis undergoing outpatient intradermal testing (IDT). Nineteen dogs were used in this clinical study. Patients were randomly allocated to receive either intravenous (IV) propofol or IV 0.9% saline, and IDT was performed on the right or left (randomized) lateral thorax. One investigator, unaware of the treatments, interpreted all IDT results. Injection sites were analysed using a subjective and objective method. A value of P or= 1+ on all dogs, significantly more positive sites were apparent during propofol sedation than during saline administration. In addition, the greater number of individual dogs experiencing more positive reactions >or= 1+ during propofol sedation was significant. When subjectively analysing reactions >or= 2+, the greater number of positive reactions and the greater number of dogs with more positive reactions observed during propofol treatment was not significantly different from the saline control. When analysed objectively, the greater number of positive reactions observed during propofol sedation was not significant. A greater number of dogs had higher subjective scores and larger objective measurements during propofol sedation compared with saline administration. In summary, propofol sedation was associated with an overall greater number of positive IDT reactions compared with the saline control. Although not always significant, this difference should be considered when choosing propofol for skin testing dogs with atopic dermatitis.     

Pilot investigation of a model for canine atopic dermatitis: environmental house dust mite challenge of high-IgE-producing beagles, mite hypersensitive dogs with atopic dermatitis and normal dogs

Although canine atopic dermatitis (cAD) is common, few models are available. The aim of this study was to evaluate high-IgE beagles epicutaneously sensitized to house dust mite (HDM) as a possible model for cAD. Six high-IgE beagles were environmentally challenged with HDM using various doses and protocols. Similar challenge protocols were used in positive and negative control dogs: three dogs with naturally occurring cAD and positive intradermal skin test (IDT) to HDM and three normal dogs without history of skin disease and negative IDT to HDM. All high-IgE beagles and all atopic dogs developed severe cutaneous lesions and pruritus after challenge. Lesions were erythematous papules and macules in contact areas such as face, ears, ventral abdomen, groin, axillae and feet. They were first visible after 6 h and increased in severity over time. No normal dog developed pruritus or lesions. Biopsies of representative lesions in the high-IgE beagles were taken for histopathology and immunohistochemistry. There was superficial perivascular dermatitis with mononuclear infiltrates and spongiosis. Lymphocytes and eosinophils accumulated in small epidermal micro-abscesses with hyperplasia of epidermal IgE-bearing dendritic cells. These findings suggest that this colony of high-IgE beagles develops a dermatitis that clinically, histopathologically and immunologically resembles the naturally occurring canine disease. It is also concluded that this modality of challenge is not irritating to normal dogs but induces flare-ups in hypersensitive atopic dogs.     

Comparison of subjective and objective intradermal allergy test scoring methods in dogs with atopic dermatitis

An intradermal allergy test (IDT) is an important diagnostic tool for identifying offending allergens in canine atopic dermatitis. No standardized method of scoring an IDT has been described. The purpose of this study was to determine whether there is a correlation between a conventional, subjective IDT scoring method based on perceived wheal diameter, erythema, and turgor (0-4+) and an objective scoring method based on measuring wheal diameter alone. Thirty-four atopic dogs were skin tested with 68 different allergens. All skin tests were performed according to standard procedures, and any IDT score ≥2+ was considered clinically significant. When the subjective IDT scores were compared with the objective IDT scores in all dogs, there was a moderate level of correlation overall (r=0.457; P <0.0001). The highest level of agreement between subjective and objective scores was noted with the reactions assigned subjective scores of "0" and "2+." Overall, there was a slight level of agreement between subjective and objective scores based on clinical significance (i.e., subjective scores ≥2+; κ=0.20; P <0.0001). In conclusion, the authors believe that the objective scoring method used in this study may provide a point of reference for inexperienced individuals (dermatology residents, veterinarians, technicians) when learning to grade an IDT.     

Response to intradermal skin testing with four cat allergen preparations in healthy and allergic dogs

Intradermal skin testing with 4 cat allergens was performed on 40 dogs. The allergens used were a commercial preparation, cat allergen 1, cat pelt extract, and cat serum. Twenty dogs had inhalant allergies (canine atopy) and 20 dogs were healthy. The dogs in these 2 groups were further allotted to groups of dogs with or without exposure to cats. Cat pelt extract was the only allergen that caused a statistically significant (P less than 0.025) difference in the number of positive reactions in healthy vs allergic dogs, with healthy dogs having the greater number of positive reactions. Seven (35%) of the 20 dogs with no known exposure to cats had positive reactions to at least one of the allergens. In dogs that had been exposed to cats, positive reactions developed in twice the number of dogs without clinical signs of canine atopy, compared with those with clinical signs of canine atopy. These data showed that dogs do become sensitized to cat allergen, but that this sensitization may not indicate known exposure to cats or clinical disease.     

Sensitivity patterns to house dust mites and forage mites in atopic dogs: 150 cases

This study investigated intradermal test reactions to extracts of six species of mites in 150 dogs with atopic dermatitis. At least one positive reaction was seen in 120 animals (80%). Dermatophagoides farinae attracted the highest number of positive reactions (108 dogs, 90% of dogs and 72% of atopic dogs showing positive reactions). Positive reactions to other mites were not uncommon, with many dogs testing positive for Dermatophagoides pteronyssinus (32% of dogs tested), Acarus siro (35%), Tyrophagus putrescentiae (30%), Glycyphagus domesticus (27%) and Lepidoglyphus destructor (23%). Sensitivity to D. farinae alone occurred commonly (57% of cases), but multiple sensitivities were seen frequently with the other mites. Cases of sensitivity to only one mite were also seen: D. pteronyssinus (five cases), T. putrescentiae (one case) and G. domesticus (one case). Further studies are needed to appreciate more clearly the precise role played by the different species of mite in canine atopic dermatitis.