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Biomedical subjects

R E Halliwell

Publications and source records attributed to R E Halliwell.

At least 19 recordsLinked to original sources

The ACVD task force on canine atopic dermatitis (III): the role of antibodies in canine atopic dermatitis.

Although an important pathogenic role for IgE is established in the case of allergic asthma and rhinitis in man, its role in atopic dermatitis is less clear. There are many studies where allergists and immunologists have provided evidence in favour of such a role, whereas dermatologists are less than convinced. In dogs, however, there is an abundance of clinical evidence implying that atopic dermatitis is antigen driven, and recent studies suggest that there may be a role for IgE, not only in the effector pathway, but also in antigen capture. Although an IgG response often accompanies an IgE response in dogs with atopic dermatitis, there is little evidence in support of a pathogenic role in respect of the former isotype.

Allergens↗

The ACVD task force on canine atopic dermatitis (XI): the relationship between arthropod hypersensitivity and atopic dermatitis in the dog.

The relationship between arthropod allergen hypersensitivity and the development of canine atopic dermatitis (AD) is unclear. It has been shown that dogs with AD are more likely to exhibit positive intradermal reactivity to flea allergens than non-pruritic dogs from the same flea-endemic geographic region. Also, dogs in a flea endemic region are four times more likely to suffer from flea allergy dermatitis (FAD) and AD than from FAD alone. These results provide indirect evidence to support the hypothesis that, in the canine species, atopy predisposes to the development of hypersensitivity to flea allergens and eventually to FAD. A causal relationship between insects other than fleas and canine AD has not been identified with certainty.

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Serum antibodies to Malassezia yeasts in canine atopic dermatitis.

Significant numbers of humans with atopic dermatitis develop Malassezia-specific IgE. Immediate skin-test reactivity to Malassezia has been demonstrated in atopic dogs. The aim of this study was to compare the serum IgG and IgE response to Malassezia in atopic dogs with and without clinical evidence of Malassezia dermatitis and/or otitis, nonatopic dogs with clinical evidence of Malassezia dermatitis and/or otitis and healthy dogs. Cytology was used to diagnose clinically significant Malassezia dermatitis and otitis. Contact plate cultures confirmed the validity of this technique. Reproducible enzyme-linked immunosorbent assays for Malassezia-specific IgG and IgE in canine serum were established. Atopic dogs had significantly higher serum IgG and IgE levels than either healthy dogs or nonatopic dogs with clinical evidence of Malassezia dermatitis and/or otitis. There was no significant difference in IgG and IgE levels between atopic dogs with and without clinical evidence of Malassezia dermatitis and/or otitis. The implications of these findings in the pathogenesis and management of canine atopic dermatitis are discussed.

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Allergen-specific IgE and IgGd antibodies in atopic and normal dogs.

Intradermal skin tests (IDSTs) were performed on 65 atopic and 24 normal dogs. The levels of allergen-specific IgE and IgGd antibodies were determined in serum samples by enzyme-linked immunosorbent assay (ELISA) using the same 12 allergens that were used in the IDST on normal dogs. The correlation between the levels of IgE and IgGd to Dermatophagoides farinae (DF) and Dermatophagoides pteronyssinus (DP) was examined. The sensitivity, specificity and positive and negative predictive values of allergen-specific IgE and IgGd levels in the total dog population were also compared. Results were consistent and reproducible for 9/12 allergens, but in the case of house dust, flea and Alternaria tenuis, a less discriminating standard curve and the fact that the negative control gave positive results, suggests non-specific binding and that these allergens are complex and should not be employed without further purification. A high percentage of atopic dogs had positive IDSTs and detectable IgE and IgGd antibodies to DF, DP and house dust. Similar results were obtained in the normal dog population. There were significant correlations between allergen-specific IgE and IgGd levels to DF and DP. However, in contrast to IgE, allergen-specific IgGd in normal dogs was higher than in atopic dogs. Furthermore, a high percentage of the atopic population had detectable IgGd to unrelated allergens, despite negative IDSTs. Overall, the negative predictive values were similar for both IgE and IgGd. Sensitivities were higher in the allergen-specific IgGd assays, but the specificities and positive predictive values were higher in the allergen-specific IgE assays. In conclusion, the concordance of IDSTs with ELISA results to DF and DP in normal dogs without clinical signs implied the possible heterogeneity of IgE in dogs. The presence of IgGd directed against apparently irrelevant allergens in atopic patients and the high levels of IgGd in normal dogs to the most common allergens, DF and DP, implied an uncertain role of IgGd in canine atopic disease. Therefore, the detection of allergen-specific IgE is a more useful adjunct to the diagnosis of atopic disease in the dog than IgGd.

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Studies on pulmonary and systemic Aspergillus fumigatus-specific IgE and IgG antibodies in horses affected with chronic obstructive pulmonary disease (COPD).

Inhalant exposure to Aspergillus fumigatus (Asp. f.) antigens induces marked inflammatory and immunological alterations in the lungs of horses affected with chronic obstructive pulmonary disease (COPD). In this study we investigated the role of specific allergen(s) present in Asp. f. on systemic and pulmonary IgE and IgG responses in control and COPD-affected horses, using an enzyme-linked immunosorbent assay (ELISA) and immunoblotting techniques. Compared with controls, horses affected with COPD had significantly higher levels of BALF IgE and IgG to somatic Asp. f. antigens as well as to the allergen 1/a (Asp. f. 1/a). Serum levels of IgE and IgG against these antigens did not differ between control and COPD-affected horses. Antigen specific IgE and IgG levels did not correlate between BALF and serum. Scanning of Asp. f. and IgE and IgG blots revealed bands that are recognised by both IgE- and IgG-specific antibodies. Additionally, all horses responded with BALF IgE- and IgG-specific for 93, 35, 31 and 23 kDa allergens, suggesting that these antigens are involved in the induction of airway IgE and IgG responses. These allergens may have the potential to be used as biomarkers for the diagnosis of Asp. f. related exacerbations of equine COPD.

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Retrospective survey of allergen immunotherapy in canine atopy.

The clinical records of 277 cases of canine atopy treated with specific allergen immunotherapy were reviewed. A good response was defined as control with immunotherapy either alone or with topical agents, a partial response as control with immunotherapy and other systemic agents, and a poor response as no perceived benefit and the immunotherapy discontinued. The mean follow-up period was 29.2 months (range 10 to 85 months). Ninety-one cases (33 per cent) were lost to follow-up or failed to comply with the therapeutic protocol. Of the remaining 186 cases, 40 (21.5 per cent) had a good response to immunotherapy, 74 (39.8 per cent) had a partial response, and 72 (38.7 per cent) had a poor response. Immunotherapy was therefore of long-term benefit in 114 dogs (61.3 per cent). No significant differences in response rates were associated with the breed or sex of the dog, or the age of onset of the disease, or with the type or number of allergens included in a vaccine. Dogs which had clinical signs for more than 61 months before immunotherapy had a significantly poorer response rate (23.5 per cent, P < 0.05). In-house cases had a significantly better response rate (95.2 per cent, P < 0.05) than externally managed cases.

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Production and characterisation of polyclonal antisera against feline IgE.

Cats, naturally or experimentally infected with Toxocara cati were immunised with dinitrophenylated ascaris antigen (DNP-Asc). All cats developed immediate skin reactivity to DNP coupled to bovine serum albumin (DNP-BSA) and the sera of the nine cats had a heat labile homocytotropic antibody detectable by homologous Prausnitz-Küstner (PK) tests. Reagin-rich fractions were prepared from these sera and used for the preparation of polyclonal antisera in rabbits. Resultant antisera were passed through a immunoabsorbent column of Sepharose 4B coupled to heated normal cat serum. An immunoabsorbent column prepared with the resultant antisera removed the PK reactivity from the cat sera, and the activity was recovered following acid elution. The antiserum failed to detect any recognised immunoglobulin in cat sera, but precipitated with a heat labile protein with gamma-1 electrophoretic mobility in the sera of parasited cats. These findings support the contention that the antisera are specific for feline IgE.

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Feline immunoglobulin E: induction of antigen-specific antibody in normal cats and levels in spontaneously allergic cats.

Sera from 10 cats with symptoms consistent with atopy, from 15 normal household cats and from 11 laboratory maintained cats were assessed for allergen-specific IgE and IgG to Dermatophagoides farinae (DF) by enzyme-linked immunosorbent assay (ELISA). In addition, 10 normal cats were immunised with DF and intradermal skin tests (IDST) were performed weekly. Sera from the latter were also assessed for DF-specific IgE by ELISA and using Prausnitz-Küstner (PK) tests. Although DF-specific IgE was detectable in all the atopic cats, there was no significant difference between the levels in this group and in the clinically normal household cats. However levels in both these groups were significantly higher than those in the laboratory maintained cats. Detectable DF-specific IgE was induced in all of the 10 cats, but the levels were not correlated with the development of positive IDSTs, nor with the level of IgE as assessed by PK tests. These findings are consistent with a possible heterogeneity of IgE antibody in cats.

Allergens↗

Canine leucocyte histamine release: response to antigen and to anti-IgE.

Histamine release from canine leucocyte-enriched peripheral blood preparations was measured in atopic, non-atopic and artificially sensitised dogs after immunological challenge with D. farinae antigen and anti-IgE. Total cell histamine and spontaneous histamine release was also measured. The total cell histamine content of equal leucocyte preparations was not statistically significant between the atopic and non-atopic groups. At all dilutions of antigen a higher amount of histamine was released from the leucocytes of atopic dogs than was seen in the non-atopic group. No histamine release in response to D. farinae was seen in the sensitised dogs although a statistically significant increase in serum D. farinae-specific IgE could be demonstrated after sensitisation (P < 0.03). Histamine release in response to anti-IgE was significantly greater in the atopic dogs than the non-atopic dogs (P < 0.004) and the sensitised dogs (P < 0.003). There was no statistically significant difference in total serum IgE between the groups. The authors conclude that the leucocytes of atopic dogs have a greater tendency to release histamine than those of normal and artificially sensitised dogs and that this is independent of the concentration of total serum IgE or antigen-specific IgE. They suggest that there may be immunoregulatory abnormalities in atopic dogs intrinsic to the atopic state as is described in man.

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Canine atopic disease: the prevalence of positive intradermal skin tests at two sites in the north and south of Great Britain.

Results of intradermal skin test responses to the same panel of 53 allergens were compared in 118 dogs with atopic disease presented at two geographical centres, Edinburgh (87 cases) and London (31 cases). The allergens most commonly positive at both centres were human dander and Dermatophagoides farinae, but positive tests to all of the allergens used occurred in at least one case. The mean number of allergens to which positive tests resulted in atopic dogs was 5.126 (Edinburgh) and 5.129 (London). The majority of animals were sensitive to allergens from more than one group. A significantly higher number of positive reactors to house dust allergen was observed at London than at Edinburgh (P < 0.05), while a significantly higher number of positive reactions to grass pollens was detected at Edinburgh than in London (P < 0.05). Sensitivity to Dermatophagoides pteronyssinus, in the absence of sensitivity to D. farinae, was uncommon and therefore both of these mite allergens should be incorporated in intradermal skin testing panels in Great Britain.

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Mast cell tryptase levels in normal canine tissues.

Levels of canine tryptase from various tissues were quantified using a competition enzyme-linked immunosorbent assay (ELISA). The assay utilises an affinity-purified rabbit anti-tryptase antibody in the solid phase and alkaline-phosphatase conjugated tryptase together with unlabelled tryptase in the fluid phase. The assay will rapidly quantify 40-5000 ng ml-1 of tryptase in tissue extracts. Tissues from the skin, gut, liver and lung were studied, of which canine gut appeared to contain the highest levels of tryptase per milligram wet weight, which may suggest an important role for this enzyme at this site. This assay may prove valuable in assessing the role of mast cells in various disease states in the dog.

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