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PubMed1981

A modified technique of guinea pig testing to identify delayed hypersensitivity allergens.

A modified guinea pig testing technique was developed for the detection of weak allergens and allergenicity of materials unsuitable for testing by intradermal injection. This test involved the use of Freund's complete adjuvant to stimulate the immune system of the animal, and external application instead of intradermal injection of the test compound in the induction stage. The allergenicity of Sudan III, Brilliant Lake Red R and Sudan I was tested by this procedure. In the dose-effect study of Sudan I, the dose dependency of a positive reaction of the induction and challenge concentrations was recognized. The test was compared with three other guinea pig sensitization tests. The results obtained with this test correlated well with those obtained with the guinea pig maximization test.

Allergens↗
PubMedDate not supplied

Anaphylaxis to oral furosemide.

Furosemide, one of the most used diuretic drugs, rarely induces type-1 allergic reactions It is included in the non-aromatic sulfonamides but a cross-reactivity mechanism between this group and the sulfonamides antibiotics, has not been clearly demonstrated. A 24-year-old woman, 10 minutes after the intake of one pill of Seguril 40mg experienced oral itching, generalized urticaria, facial angioedema, dyspnea and hypotension. She recovered after the administration of parental adrenaline, methyl-prednisolone and dyphenhydramine. An skin prick test with furosemide (10 mg/ml) was negative. The intradermal skin tests were positive to furosemide (1 %) as well as sulfamethoxazole (0.03 mg/ml), with 10 atopic and non-atopic negative controls. The patient rejected the performance of an oral challenge test with sulfamethoxazole. IgE-mediated reactions to furosemide are infrequent, but it could be the cause of life-threatening reactions. We have reported a case of anaphylaxis after the oral administration of furosemide with a demonstrated hypersensitivity mechanism through the positive intradermal skin test. The previous administration of the drug could probably the mechanism of sensitization, but the positive intradermal test to sulfamethoxazole would open the hypothesis of a cross-reactivity between non-aromatic and antimicrobial sulfonamides. It could be necessary an oral challenge test with furosemide in allergic patients to sulfamides.

Administration, Oral↗
PubMed1997

Skin tests in the diagnosis of eruptions caused by betalactams.

The aetiologic evaluation of adverse cutaneous reactions to penicillins is still not an easy problem to solve. Skin testing is usually carried out intradermally with benzylpenicilloyl polylysine (BPO-PPL) and minor determinant mixture (MDM), but these are often unsuitable for the detection of sensitivity to betalactam antibiotics. 101 selected subjects, with different cutaneous reactions to betalactams and with a clinical history of positive challenge, were skin tested (patch test, prick test, intradermal test) with a standard betalactam series (amoxycillin, sodium penicillin G, ampicillin, bacampicillin, aztreonam, ceftriazone, BPO-PPL, MDM). 1 or more positive reactions to skin tests, mainly to intradermal tests, were observed in 47.5% of the subjects studied, especially in those with maculopapular eruptions, urticaria/angioedema and drug reactions caused by ampicillin and amoxycillin. Cross-sensitivity was demonstrated in 22.8% of cases and was due almost solely to the semisynthetic penicillins. Finally, to increase the yield in detecting positive patients, it is necessary that ampicillin and amoxycillin be tested in addition to major and minor determinants.

Adolescent↗
PubMed2001

Chronic urticaria: novel clinical and serological aspects.

BACKGROUND: Recently, distinct studies have shown that: (a) chronic idiopathic urticaria (CIU) is autoimmune in 30-50% of cases; (b) in patients with CIU the autologous serum skin test is inhibited by heparin; and (c) basophil histamine release induced in vitro by CIU sera maybe complement-dependent. OBJECTIVE: To carry out a comprehensive clinical and serological study on CIU based upon these observations. METHODS: Three hundred and six adults with CIU underwent intradermal (ID) test with autologous serum; 57 of them with autologous heparinized plasma as well. Sera from 121 patients (plasmas from 17) were employed to induce in vitro histamine release from basophils of normal donors. The effects of heating (56 degrees C, 60 min), filtration through membrane, and preincubation with heparin were evaluated as well. RESULTS: Autologous serum and plasma induced a weal and flare reaction in 205 out of 306 (205/306; 67%) and in 8/57 (14%) patients, respectively. Positive plasma skin tests were observed only in patients showing strongly positive serum skin tests. Plasma did not elicit any skin reaction in 3/3 patients with dermatographism who showed a positive intradermal test with saline. Sera from 20/121 (16.5%) patients induced significant histamine release from basophils of normal donors. 19/20 sera were from patients with a positive intradermal test; thus, basophil histamine release assay was positive in 19/87 (21.8%) patients with a positive serum skin test. Heating at 56 degrees C x 1 h markedly reduced the histamine-releasing activity of both serum and plasma from in vitro reactors. Ultrafiltered fractions > 100 kDa of both sera tested retained the histamine-releasing activity, whereas fractions < 100 kDa were not able to induce any histamine release. Heparin dose-dependently inhibited histamine release induced by sera and plasma, and by basophil agonists such as anti-IgE, formyl-methionyl-leucyl-phenilalanine, and interleukin (IL)-3. CONCLUSIONS: 67% of our patients with CIU showed a positive autologous serum skin test. Sera from about 20% of those positive on autologous serum skin test induced histamine release from normal basophils in vitro probably as a consequence of the presence of functional autoantibodies. The marked difference between in vivo and in vitro findings could reflect the existence of a mast cell-specific histamine-releasing factor which does not release histamine from basophils of healthy blood donors. However, it might be also the result of in vivo priming of patients' cutaneous mast cells or of heterogeneity of basophil donors. At least in some cases complement seems essential for histamine-releasing activity of serum from patients with CIU. Heparin inhibits histamine release from both basophils (in vitro) and mast cells (in vivo), probably acting directly at a cellular level.

Adolescent↗
PubMed2001

Natural rubber pharmaceutical vial closures release latex allergens that produce skin reactions.

BACKGROUND: The release of allergenic proteins from natural rubber vial closures (stoppers) into aqueous pharmaceuticals may induce allergic reactions in individuals with latex allergy (LA) receiving medications from such vials. OBJECTIVE: The goal of this study was to determine whether solutions stored in vials containing natural rubber closures release allergenic proteins detectable by skin testing of subjects with LA. METHODS: Five pharmaceutical vial closures (2 natural rubber and 3 synthetic) were coded, inserted onto vials containing phenol-saline-human serum albumin, and stored in an inverted position before use. Twelve volunteers with and 11 volunteers without LA underwent skin testing with solutions from each of the 5 vials, either those not punctured (0P) or those punctured 40 times with a 21-gauge needle 12 to 24 hours before testing (40P). RESULTS: All intradermal skin test responses in the group without LA were negative. Two and 5 of the 12 subjects with LA had positive intradermal skin reactions to 0P and 40P solutions, respectively, from vials containing rubber closures. Two subjects with LA had inexplicable, positive, nonreproducible intradermal skin test reactions to solutions from vials containing bromobutyl but not vials with isoprene synthetic closures. In vitro inhibition analysis detected 6 to 7 AU/g latex allergen in extracts of cut natural rubber containing closures but not in extracts of synthetic closures. CONCLUSION: Natural rubber vial closures released allergenic latex proteins into the tested solutions in direct contact during storage in sufficient quantities to elicit positive intradermal skin reactions in some individuals with LA. These data support a recommendation to eliminate natural rubber from closures of pharmaceutical vials.

Adolescent↗
PubMed1996

[Tests for atopic antigens].

It is important to determine what kind of antigen is involved in atopic asthma for appropriate management as well as diagnosis. We can measure total IgE and antigen specific IgE antibodies by means of RIA and EIA in vitro. Antigen specific IgE antibodies can be detected more sensitively by skin tests consisting of intradermal, prick and scratch tests, among which the intradermal test is most sensitive. To study if the antibody thus detected is functional, there are in vivo tests such as the provocation test and eye reaction with the corresponding antigen and in vitro tests such as the histamine release assay and CAST. In Japan we tend to prefer in vitro assays, while the skin test has been recommended more than in vitro assays in the United States.

Asthma↗
PubMed1994

Hypersensitivity to therapeutic murine monoclonal antibodies.

OBJECTIVE: To determine the predictive value of pre-treatment skin tests and in vitro IgE/IgG4 anti-murine monoclonal antibodies in patients treated with murine monoclonal antibodies. DESIGN: Patients treated at two cancer institutions were evaluated by skin testing and solid phase immunoassays to detect IgE and IgG4 specific anti-murine monoclonal antibodies. Skin testing by scratch and intradermal skin testing was done on patients before treatment with murine monoclonal antibodies. IgE & IgG4 specific anti-murine monoclonal antibodies were determined before treatment in all patients and at 1, 7, 14 and 21 days post-treatment in 1 patient. SETTING: Cancer patients undergoing murine monoclonal antibody treatment in two university medical centers were recruited for the study. PARTICIPANTS: Twelve patients, aged 41-75 years with gastrointestinal cancers (colon, stomach, pancreas or liver) with metastatic disease, who had relapses or conventional therapy were enrolled. Some patients had previous exposure to rodents, either as laboratory personnel or had kept them as pets. INTERVENTION: One patient who experienced an anaphylactic reaction to murine monoclonal antibody infusion was desensitized so therapy could continue. MAIN OUTCOME MEASURES: Skin tests, immunoassays, and patient history were correlated with adverse reactions to infusions of murine monoclonal antibodies. MAIN RESULTS: Skin tests (scratch method) and/or in vitro immunoassays may predict allergic outcomes in patients receiving infusions of murine monoclonal antibodies. Intradermal skin testing with murine monoclonal antibodies may result in false positive reactions and have less predictive value. Specific IgE or IgG4 were elevated in the two patients who experienced severe adverse reactions to murine monoclonal antibodies but not in those patients with no reactions and therefore, may have some predictive value. A history of past exposure to mice may also increase the risk of adverse reactions. In one patient, intravenous desensitization enabled treatment to proceed. CONCLUSION: Scratch skin tests, in vitro IgE and/or IgG4 immunoassays together with a past history of previous exposure to murine antigen(s) may predict potential allergic reaction to therapy with murine monoclonal antibodies.

Adult↗
PubMedDate not supplied

Preparation of benzylpenicilloyl-polylysine: a preliminary study.

The manufacturing of PrePen (benzylpenicilloyl-polylysine) was suspended from September 2000 through November 2001 because of Food and Drug Administration intervention. PrePen is penicilloyl bound to polylysine and is considered to be the major determinant of penicillin metabolism; it is used as a skin-testing reagent to detect immunoglobulin E antibodies in people with a history of penicillin allergy. Because of the uncertainty of whether PrePen would be made available in the future, we attempted to make benzylpenicilloyl-polylysine in our laboratory using a modification of a previously described method. Subsequently, we compared our preparation with PrePen by enzyme-linked immunosorbent assay and skin testing. Using a patient previously documented to have a positive reaction to PrePen by history and intradermal skin testing, a puncture test to PrePen as well as dilutions of benzylpenicilloyl-polylysine was negative. Positive intradermal tests were performed on PrePen and benzylpenicilloyl-polylysine at 5 mg/mL. The enzyme-linked immunosorbent assay was performed to compare the antigenicity of PrePen and benzylpenicilloyl-polylysine and revealed similar binding to both. Subsequently, cutaneous testing on 10 additional penicillin-allergic people was performed, with corroborative results. Our benzylpenicilloyl-polylysine production methodology could prove useful to those who do not have access to PrePen or another similar reagent.

Benzeneacetamides↗
PubMed1986

Skin tests in bird breeder's disease.

The diagnostic value of skin tests was assessed in 35 patients affected by bird breeder's disease by comparing the results with those obtained in a group of 27 symptomless breeders and 10 symptomless unexposed individuals. Sterilised and diluted serum and lyophilised extracts of faeces prepared from the breeder's own birds were injected intradermally. Skinprick tests using the same materials were carried out simultaneously. The intradermal test using serum showed a positive reaction in 18 out of 20 patients tested and in three out of 20 symptomless breeders (p less than 0.0005); later reactions occurred in eight of the patients and one of the symptomless breeders (p less than 0.01). A delayed reaction was seen in five of the patients and one of the symptomless breeders (p = 0.09). No positive reactions were recorded in symptomless unexposed individuals. Intradermal skin testing using extracts of faeces yielded an immediate reaction in 19 of 21 patients and in three of 11 symptomless breeders tested (p less than 0.001); a late reaction was seen in 18 of the patients and in five of the symptomless breeders (p less than 0.05). Delayed reactions occurred in 11 of the 21 patients tested, whereas no positive responses were seen in symptomless breeders (p less than 0.025). Skinprick tests gave negative results in all cases. Skin testing constitutes a simple, quick, and safe procedure that can assist the differential diagnosis between individuals with bird breeder's disease and symptomless breeders, the immediate reaction being similar in its sensitivity to testing for serum precipitins but possessing greater specificity.

Adolescent↗
PubMed2001

Recombinant allergens for skin testing.

Skin testing is a basic diagnostic procedure widely used to explore immediate-type reactions to allergen preparations in vivo. Despite their reliability, if standardized extracts are used, skin tests suffer from limited reproducibility due to difficulties in preparing consistently standardized extracts from natural raw material. Starting from allergen-encoding cDNAs, large amounts of highly pure allergens with a high batch-to-batch consistency satisfying the quality requirements of medicinal products manufactured by recombinant DNA technology can be produced. These reagents are expected to be qualitatively superior to the commercially available allergen preparations used for the in vitro and in vivo diagnosis of allergic conditions. In this article the current literature available on skin testing with such recombinant allergens (rAllergens) is reviewed and critically analyzed. To date many different rAllergens of various pollens, moulds, mites, bee venom, latex and celery have been used in skin testing in more than 1,600 allergic and control individuals. Skin prick tests as well as intradermal skin tests with rAllergens prove to be highly specific and safe. The diagnostic sensitivity of single rAllergens is generally lower than those obtained with allergen extracts, but can be considerably increased by using rAllergen panels covering the most important allergenic structures present in a given complex allergenic extract. Moreover, quantitative skin testing with single rAllergens allows interesting insights into correlations between the in vivo and in vitro sensitization to a given allergen. In conclusion, skin testing with rAllergens offers a highly specific and safe additional diagnostic tool to elucidate patient- and disease-specific sensitization patterns which will be needed for the development of patient-tailored immunotherapeutic treatments.

Allergens↗
PubMed2000

Effective DNA vaccination of cattle with the mycobacterial antigens MPB83 and MPB70 does not compromise the specificity of the comparative intradermal tuberculin skin test.

The current tuberculin test and slaughter strategy for the control of bovine tuberculosis in cattle has failed to prevent a sharp rise in cases over recent years, especially in the south-west of England. A recent scientific review has concluded that the development of a cattle vaccine holds the best prospect for tuberculosis control in British herds. In order to continue with test and slaughter-based control strategies, the development of TB vaccines that do not compromise the specificity of the tuberculin skin test are required. This report describes results of cattle vaccination experiments with TB DNA vaccines expressing the mycobacterial antigens MPB70, MPB83, and Ag85A and constitutes the first published vaccination study with DNA vaccines undertaken in a target host species. All calves vaccinated with the MPB83 expressing plasmid demonstrated potent cellular immune responses, characterised by CD4(+) T cells producing interferon-gamma as well as humoral immunity characterised by IgG1 biased specific antibodies. Vaccination with MPB70 was less effective with immune responses only observed in half of the vaccinated animals, while vaccination with Ag85A did not result in vaccine-induced immune responses. Intramuscular vaccination was found to stimulate stronger cellular responses than intradermal immunisation. Significantly, the specificity of tuberculin skin testing was not compromised by DNA vaccination since none of the vaccinated calves showed positive skin test reactivity.

Administration, Cutaneous↗
PubMed2002

Extracts of Anacardium occidentale (cashew) pollen in patients with allergic bronchial asthma.

Allergic reactions to the pollen of trees is among the most prevalent allergic sensitivities. The cashew tree grows in abundance in the northeast region of the Brazil, mainly in Fortaleza city, in state of the Ceará. It flowers once a year between August and October. This is the first study conducted to establish the possible role of the cashew pollen extract in causing skin test reactivity in patients with allergic asthma. A stock solution of pollen extract was prepared with the standard weight/volume method for intradermal skin tests and for the protein content of the extract, estimated with the use of Folin phenol reagent and a spectrophotometer. Ten nonallergic volunteers and 80 subjects with allergic asthma, as documented by previous positive skin test reactions to various pollens, were studied. All of the 80 patients (100%) had positive test reactions (grade III and grade IV reactions). None of the control subjects (n = 10) had positive responses to the intradermal tests. This study provided us with knowledge of an additional pollen extract of the Anacardium occidentale, which could provoke skin test reactivities in asthmatic individuals from the northeastern area of Brazil. The results suggest a relationship between the period of flowering of the cashew tree and the increased number of allergic asthma cases.

Adolescent↗
PubMed2003

The prevalence of immediate and delayed type hypersensitivity reactions to Microsporum canis antigens in cats.

Spontaneous recovery from Microsporum canis infections in cats is thought to be dependent on the development of a competent immune response. The purpose of this study was to determine the prevalence of positive delayed type hypersensitivity reactions in cats with and without dermatophytosis. Four groups of cats were intradermally skin tested with M canis extract and test sites were evaluated both subjectively and objectively at 0, 24 and 48 h after injection. Delayed intradermal testing (IDT) reactions were absent in cats not exposed to dermatophytosis (n=20); infected-recovered cats (n=38 culture negative lesion negative and n=43 lesion negative but culture positive) had significantly larger IDT reactions than unexposed cats and cats that were still actively infected (n=18). Based on the results of this study, IDT with M canis extract can be used to assess the cellular immune response of cats with dermatophytosis.

Animals↗
PubMed1977

Specific and non-specific skin test reactions with animal dander extract.

Intradermal skin tests have been performed in atopic patients with allergenic extracts of different animal danders at uniform concentration. A high incidence of positive test was recorded which could not be related to the clinical histories, especially with the allergens of horse, cow and dog dander (NR). Better correlation was observed with guinea pig and cat allergens (R). The incidence of NR reactions was highest in the house dust allergy group of patients; with the exception of horse dander allergen, the frequency of R reactions was more pronounced in the grass pollen group. It is suggested that both allergen-specific and non-allergen-specific mechanisms may contribute to positive wheal and flare reactions.

Allergens↗
PubMed1973

Cell-mediated immune response in cattle to Mycoplasma mycoides var. mycoides.

The cell-mediated immune response of cattle to Mycoplasma mycoides var. mycoides was studied. Sensitized lymphocytes in blood leukocyte preparations showed a significant degree of antigen-induced transformation, judged by the uptake of tritiated thymidine. The increase in tritiated thymidine uptake in sensitized lymphocytes in the presence of M. mycoides membrane antigen varied from 2- to 13-fold compared with the controls, and this increase in activity was observed from 3 days after artificial infection. Inhibition of leukocyte migration by M. mycoides membrane antigen commenced between 17 and 30 days after infection, and preliminary observations indicate that this test correlated with the intradermal allergic test. M. mycoides-induced unresponsiveness was demonstrated 23 and 30 days after infection. Unresponsiveness, in that the lymphocytes did not respond to phytohemagglutinin, was very marked in two of three animals and partial in the third animal, whereas the humoral antibody response did not appear to be affected. Antigen-induced transformation was demonstrated in only two out of six cattle vaccinated two months previously with M. mycoides T(1) broth culture vaccine, and óne animal only gave a doubtful intradermal allergic reaction. A further six cattle vaccinated 15 months previously were negative to both the leukocyte migration inhibition test and the intradermal allergic test.

Animals↗