Characterization of the sensitization profile of bee venom allergic patients ­ Association with the severity of reaction? / Caracterização do perfil de sensibilização molecular de doentes alérgicos à picada de abelha - Associação com a gravidade da reação?

Introduction: Bee venom (BV) allergy, a common cause of anaphylaxis in adults, is often associated with severe reactions. The use of component-resolved diagnostics (CRD) increases diagnostic accuracy. Objectives: To characterize the sensitization profile of BV allergic patients and a possible correlation with the severity of reaction. Materials and methods: We selected patients with a clinical history of BV allergy, positive skin tests, and specific IgE (sIgE) for BV. The allergenic profile was analyzed by both CRD and Western blot using a well-defined and properly characterized BV extract. Results: Forty-four patients were included, 30 (68.2%) were men. Mean age was 48.9 (SD 17.9) years. Eleven (25%) had large local reactions (LLRs) and 33 (75%) had systemic sting reactions (SSRs). One patient with negative sIgE for BV had positive sIgE for Api m 1, Api m 5, and Api m 10. The sensitization frequency for BV, Api m 1, Api m 2, Api m 3, Api m 5, and Api m 10 was 97.7%, 75%, 47.7%, 20.5%, 40.9%, and 61.4%, respectively. Five patients (11.4%) were sensitized to all BV components. CRD association showed that 5 patients (11.4%) were sensitized only to Api m 1, 8 (18.2%) to Api m 1/Api m 3/Api m 10, and 16 (36.6%) to Api m 1/ Api m 10. Twenty-eight patients (84.8%) with SSRs were sensitized to Api m 1, and concomitant sensitization to Api m 1/Api m 10 was detected in 20 (60.6%). There was a significant difference in Api m 1 between patients with LLRs and SSRs (p = 0.0104). Similar profiles were identified by Western blot analysis, with relevance for the detection of Api m 6 in 28 (64%) and Api m 4 in 16 (36%) patients. Conclusion: The analysis of the sensitization profile using CRD and the association of several of these components can increase diagnostic accuracy in BV allergy. Our data showed that concomitant sensitization to Api m 1 and Api m 10, detected by both CRD and electrophoretic profile, may be associated with SSRs. We emphasize the identification of sensitization to Api m 6 in > 50% of patients, which may be considered a major allergen, and to Api m 4, which may be related to reactions during BV immunotherapy.

Introdução: A alergia ao veneno de abelha (VA) é uma causa frequente de anafilaxia em adultos e está muitas vezes associada a reações graves. O diagnóstico por componentes moleculares (CRD) contribui para uma melhor caracterização desta alergia. Objetivos: Caracterização do perfil de sensibilização molecular de doentes alérgicos ao veneno de abelha e possível correlação com a gravidade da reação. Material e métodos: Selecionaram-se doentes com história de alergia a VA, testes cutâneos e IgE específica (sIgE) positivos para VA. Avaliou-se o perfil alergênico por CRD e por Western Blot, utilizando extrato de VA bem caracterizado. Resultados: 44 doentes, 30 (68,2%) sexo masculino. Média de idades 48,9 ± 17,9 anos, 11 (25%) com reacções locais exuberantes e 33 (75%) com reações sistêmicas à picada (SSR). Um doente tinha sIgE negativa para VA, mas Api m 1, Api m 5 e Api m 10 positivas. A frequência de sensibilização para VA, Api m 1, Api m 2, Api m 3, Api m 5 e Api m 10 foi 97,7%; 75%; 47,7%; 20,5%; 40,9% e 61,4%, respectivamente. Cinco (11,4%) doentes estavam sensibilizados a todos os componentes. Por associação de CRD, detectaram-se 5 (11,4%) doentes sensibilizados apenas a Api m 1, 8 (18,2%) a Api m 1/Api m 3/Api m 10, e 16 (36,6%) a Api m 1/Api m 10. Vinte e oito (84,8%) doentes com SSR tinham Api m 1 positiva e 20 (60,6%) tinham Api m 1/Api m 10 simultaneamente positivas. Observou-se uma diferença estatisticamente significativa para a Api m 1 entre doentes com reações locais exuberantes e sistêmicas (p = 0,0104). Os perfis detectados por Western Blot foram semelhantes, de referir, à detecção de Api m 6 em 28 (64%) e Api m 4 em 16 (36%) dos doentes. Conclusão: A análise do perfil de sensibilização através de CRD e a sua associação aumentam a precisão do diagnóstico de alergia a VA. Sensibilização simultânea a Api m 1 e Api m 10 identificados tanto por CRD como por perfil eletroforético, pode estar associada à ocorrência de SSR. Destaca-se a sensibilização a Api m 6 em > 50% dos doentes, podendo ser considerado um alergênio major, e a Api m 4, possivelmente associado a reações durante a imunoterapia com VA.

Local inflammatory reaction induced by Scolopendra viridicornis centipede venom in mice.

Centipede envenomation is generally mild, and human victims usually manifest burning pain, erythema and edema. Despite the abundance and ubiquity of these animals, centipede venom has been poorly characterized in literature. For this reason, the aim of this work was to investigate local inflammatory features induced by Scolopendra viridicornis centipede envenomation in mice, evaluating edema formation, leukocyte infiltration, production of inflammatory mediators, and also performing histological analysis. The highest edematogenic activity induced by the venom, determined by plethysmometry, was noticed 0.5 h after injection in mice footpad. At 24 h, edema was still detected in animals that received 15 and 60 µg of venom, and at 48 h, only in animals injected with 60 µg of venom. In relation to leukocyte count, S. viridicornis venom induced cell recruitment, mainly neutrophils and monocytes/macrophages, in all doses and time periods analyzed in comparison with PBS-injected mice. An increase in lymphocytes was detected especially between 1 and 24 h at 60 µg dose. Besides, eosinophil recruitment was observed mainly for 15 and 60 µg doses in early time periods. Edema formation and cell recruitment were also confirmed by histological analysis. Moreover, S. viridicornis venom stimulated the release of IL-6, MCP-1, KC, and IL-1ß. Conversely, S. viridicornis venom did not induce the release of detectable levels of TNF-α. We demonstrated that the edematogenic activity induced by S. viridicornis venom was of rapid onset, and the venom stimulated secretion of pro-inflammatory mediators which contribute to the inflammatory reaction induced by S. viridicornis venom in an experimental model.

Avaliação da toxicidade e da resposta cutânea local induzidas por doses diluídas de veneno de abelha em cães

O veneno de abelhas (BV, do inglês beevenom), vem sendo usado com propósitos terapêuticos tanto em medicina humana quanto em medicina veterinária. A apipuntura é uma prática terapêutica da acupuntura, onde o veneno de abelhas é injetado empontos de acupuntura, através do próprio ferrão do inseto na pele ou de aplicação de injeções de veneno diluído. Apesar dos efeitos promissores do BV, o potencial tóxico do veneno de abelhas deve ser considerado. O objetivo deste estudo foi monitorar os possíveis efeitos tóxicos e a resposta cutânea local induzida pela injeção de doses diluídas em cães Beagles sadios. A resposta cutânea local foi mensurada através da distensão tecidual relativa onde a distensão da orelha esquerda, injetada com 0,1 ml de uma solução de BV diluído em salina nas doses de 0,3mg/kg (n=4), 0,043mg/kg (n=4) de BV/animal ou de salina (grupo controle, n=4) foi comparada com a distensão da orelha direita injetada com salina. A distensão tecidual relativa foi analisada nos tempos 5, 10, 15, 30 minutos, 1, 6, 12, 24, 48 e 72 horas pós inoculação do veneno. As alterações clínicas como freqüência cardíaca e respiratória, pressão arterial e temperatura; assim como as reações comportamentais; os perfis hematológico (hemograma) e renal (uréia, creatinina) e foram monitorados em diferentes tempos após a inoculação. O pico de distensão tecidual relativa ocorreu 6 horas após a inoculação do veneno, sendo a dose de 0,3mg/kg capaz de produzir distensão diferente do controle-salina nos tempos 1, 6 e 12 horas após inoculação. Não foram detectadas diferenças siginificativas entre os grupos nos demais parâmetros analisados. Estes resultados indicam que o veneno de abelhas em cães produz um pico dereação cutânea local 6 horas após a inoculação e que as doses de 0,3mg/kg e 0,043mg/kg não produzem alterações comportamentais ou autonômicas significaticas além de não produzir nefrotoxidade ou alterações no perfil hematológico de cães.

The bee venom (BV) has been used for therapeutic purposes in both human medicine and veterinary medicine. The apipuncture is a therapeutic practice of acupuncture, where the bee venom is injected into acupuncture points, by their own insect's sting into the skin or injections of diluted venom. Despite the promising effects of BV, the toxic and allergenic potential of bee venom should be considered. The aim of this study was to check the possible toxic effects and local cutaneous reaction of the injection of diluted doses in healthy dogs Beagles. The local cutaneous reaction was measured by relative tissue distension on where the distension of the left ear (injected with 0.1 ml of a solution BV diluted with saline in the doses of 0.3 mg / kg (n = 4), 0.043 mg / kg (n= 4) BV / animal or saline (control group, n = 4) was compared with the distension of the right ear, injected with saline. The tissue distension was analyzed on the times of 5, 10, 15, 30 minutes, 1, 6, 12, 24, 48 and 72 hours post inoculation. Clinical changes such as heart rate and respiratory rate, blood pressure and temperature as well as behavioral responses, the profiles hematologic (blood) and kidney (urea, creatinine and urinalysis) were monitored at different times after inoculation. The peak of tissue distension occurred 6 hours after venom inoculation and the dose of 0.3 mg/kg producedsignificant higher distension than the saline-control group one, 6 and 12 hours after inoculation. No siginicant differences were found between groups for the other parameters. These results indicate that the bee venom in dogs produced a peak of cutaneous distension 6 hours after inoculation and that the dosis of 0.3 mg / kg and 0.043 mg / kg did not produced significant behavioral or autonomic changes and did not produced nephrotoxicity or hematological changes in dogs.

Inflammatory mediators generated at the site of inoculation of Loxosceles gaucho spider venom

Patients bitten by Loxosceles spiders generally manifest marked local inflammatory reaction and dermonecrosis. This report evaluated edema formation, leukocyte infiltration and release of inflammatory mediators at the injection site of Loxosceles gaucho venom. BALB/c mice were i.d. injected with venom and thereafter paws were disrupted and homogenizedto obtain differential counts of migrated cells, as well to assay the levels of cytokines, chemokines and lipid mediators. Increased footpad thickness was detected as soon as30 min after venom injection, and 24 h later was similar to that of the control group. Loxosceles venom mildly augmented the recruitment of leukocytes to the footpad in comparison with PBS-injected mice. Moreover, it stimulated the release of IL-6, MCP-1 and KC at 2 and 24 h after venom injection. In addition, higher levels of PGE2 were detected30 min after venom injection in comparison with control group. However, the venom failed to increase levels of IL-1b, TNF-a, TXB2 and LTB4. Our results demonstrate that L. gaucho venom evokes an early complex inflammatory reaction, stimulating the secretionof pro-inflammatory cytokines and lipid mediators (PGE2), and recruiting leukocytes to the $footpad which contribute to the local reaction induced by L. gaucho venom.