Allergenicity and cross-reactivity of Senecio pollen: identification of novel allergens using the immunoproteomics approach.

BACKGROUND: The genus Senecio is the largest genus of the family Asteraceae (Compositae). The allergenicity of Senecio has not been assessed previously. OBJECTIVE: The aim of this study was to investigate the allergens of Senecio jacobea pollen and to determine their immunological characteristics and clinical relevance. METHODS: Fifty patients with rhinoconjunctivitis and a positive skin prick test (SPT) to Senecio were recruited. The clinical relevance of this pollen was assessed by means of a nasal provocation test (NPT). Allergens were characterized by one-dimensional electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis and immunoblotting. Furthermore, characterization and identification of the allergens were performed by mass spectrometry (MS). In vitro inhibition tests were performed to evaluate cross-reactivity with other pollen. RESULTS: Three predominant allergens, both in the intensity of reaction and the frequency of recognition by human-allergic sera, were 59 (60%), 42 (50%) and 31 kDa (50%). The two-dimensional analysis allowed the identification of several allergens. One spot around 42 kDa was identified as a protein homologous to pectate lyase and three other spots were homologous to malate dehydrogenase by MS. S. jacobea proteins showed cross-reactivity with other proteins of the Asteraceae family and also with Parietaria judaica. This was demonstrated by immunoblotting and ELISA inhibition studies. CONCLUSION: S. jacobea constitute a newly discovered allergenic source. It shows cross-reactivity with other members of the Asteraceae plant family as well as with P. judaica.

Associations among pollen sensitizations from different botanical species in patients living in the northern area of Madrid.

OBJECTIVE: To determinate the existence of associations among sensitizations to antigens produced by pollen grains of different botanical species as assessed by skin prick tests in patients with respiratory disorders. METHODS: Six hundred twenty nine consecutive patients living in the northern area of Madrid who underwent clinical evaluation because of rhinoconjunctivitis, and/or asthma were studied. All patients were tested with a skin prick test using a battery of inhalants including pollens, dust mites, molds and danders. The exploratory multivariate technique of Multiple Correspondence Analysis was used to compare the homogeneity of sensitizations between groups. Of the 629 patients, 459 (73.0%) had positive skin prick tests to pollen and were selected as the study group. RESULTS: The most prevalent pollen sensitization was to Gramineae pollen (83.7%) followed by Oleaceae sensitisation (75.8%). Multiple Correspondence Analysis revealed the existence of an association among pollen sensitizations, showing that they clustered two groups: sensitizations to Gramineae, Oleaceae, Cupressaceae, Chenopodiaceae, Plantaginaceae (group I), and sensitizations to Betulaceae, Platanaceae, Compositae (group II). Sensitization to Parietaria was not included in any of the sensitization groups and showed an independent behaviour. CONCLUSION: Pollen sensitizations in our area cluster into two association groups which have not previously been reported.

Cross-reactivity between Parietaria pollen and beet

Allergy to beet is very rare. Until now, only a few reports about asthma induced by inhaling the vapor of cooked beet have been published. We describe two patients with allergic rhinitis and positive skin prick tests to Parietaria and beet only. To investigate possible cross-reactivity between Parietaria pollen and beet, we performed laboratory assays that showed beet-specific IgE in the sera of both patients and possible cross-reactivity between Parietaria and beet in one patient

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Randomized open comparison of the safety of SLIT in a no-updosing and traditional updosing schedule in patients with Parietaria allergy.

BACKGROUND: Due to optimal safety of sublingual immunotherapy (SLIT), it was suggested that a slow up-dosing phase maybe not necessary, and therefore the treatment will be more patient-friendly, avoiding dosing mistakes. PATIENTS: Twenty adult patients suffering allergic rhinitis due to Parietaria, were enrolled. Half of them received the traditional schedule and the other half immediately started with 200 STU. RESULTS: No difference was observed between the traditional up-dosing treatment schedule and no-up-dosing treatment schedule in terms of side effects, even mild local side effects, even mild local side effects was greater with traditional regimen.

Age, gender and reactivity to allergens independently influence skin reactivity to histamine.

BACKGROUND: The ability to mount an IgE response to allergens is a prerequisite for the development of positive allergen skin tests. Histamine is commonly used as a positive control in skin prick testing and provides a measure of nonspecific skin reactivity, similar to bronchial hyper-responsiveness. METHODS: To determine whether allergen responsiveness, age, gender and season of the year contribute to histamine sensitivity, 620 subjects (502 of them with at least one known sensitizing allergen and the remaining 118 non-allergic controls) were prick-tested with a panel of allergens common in the Northern Italy semi-rural area where the patients lived, and with 10 mg/ml histamine dihydrochloride. RESULTS: We found higher histamine reactivity in allergic versus control individuals (median value 23.7 versus 19.8 mm2; p=0.0497). Likewise, we found in allergic subjects a correlation between allergen responsiveness in terms of number of positive allergens at skin prick test and sensitivity to histamine (mono- sensitized versus poly-sensitized subjects: p=0.0015). Moreover older age and male sex were associated with a higher response to histamine, also when separately considering allergic subjects (p<0.0001 in both cases: correlation coefficient for age versus histamine reactivity: r=0.3408). The correlation between allergen responsiveness and sensitivity to histamine was maintained also when statistically balanced for age and sex. CONCLUSION: Allergen responsiveness, gender and age allow more accurate prediction of histamine sensitivity than either parameter alone.

Randomized open comparison of the safety of SLIT in a no-updosing and traditional updosing schedule in patients with Parietaria allergy / Estudio comparativo abierto y aleatorio de la seguridad de la inmunoterapia sublingual con una pauta no progresiva y otra progresiva tradicional en pacientes alérgicos a parietaria

Background: Due to optimal safety of sublingual immunotherapy (SLIT), it was suggested that a slow up-dosing phase maybe not necessary, and therefore the treatment will be more patient-friendly, avoiding dosing mistakes. Patients: Twenty adult patients suffering allergic rhinitis due to Parietaria, were enrolled. Half of them received the traditional schedule and the other half immediately started with 200 STU. Results: No difference was observed between the traditional up-dosing treatment schedule and no-updosing treatment schedule in terms of side effects, even mild local side effects was grater with traditional regimen

Antecedentes: Debido a la óptima seguridad de la inmunoterapia sublingual, se sugirió que la fase de aumento progresivo de la dosis podría no ser necesaria, y por lo tanto el tratamiento sería más fácil para los pacientes, evitándose los errores de dosificación. Pacientes: Se seleccionaron 20 pacientes adultos con rinitis alérgica por sensibilización a Parietaria. La mitad de ellos recibió la pauta tradicional y la otra mitad empezó con la dosis de 200 STU. Resultados: No ser observó diferencia entre la pauta tradicional y la que se inició con la dosis de mantenimiento, en relación con efectos colaterales, incluso hubo efectos locales más frecuentes con el régimen tradicional

Aeroallergens sensitization in an allergic paediatric population of Cova da Beira, Portugal

Cova da Beira is an interior central region of Portugal, with a population of 93 000 inhabitants. The first pollen counts performed in Portugal revealed the highest values of the country in this area. The aim of this study was to assess the aeroallergens sensitization in an allergic population, according to the age groups. In a 5 year period (1995-2000) 1790 consecutive outpatients were observed for suspected allergic symptoms. We included in this study all the 557 paediatric (<= 15 years old) observed patients (317 male (57 %) and 240 female (43 %) with an average age of 7.6 <= 4.2 years old). They were divided in three age groups (Group I: <= 5yr; Group II: 6-10yr; Group III: 11-15yr). 371 patients were submitted to skin prick tests to aeroallergens. 86.5 % of these patients were sensitised to at least one allergen extract. The most representative aeroallergens sensitization were grasses mixture (44.9 %), D. pteronyssinus (32.5 %), D. farinae (29.1 %), Olea europea (27.5 %), Parietaria judaica (23.4 %), cat dander (16.1 %), Artemisia vulgaris (17.6 %), Robinia pseudoacacia (12.2 %), Platanus acerifolia (11.4 %), Tilia cordata (11.4 %) moulds mixture (11.2 %), Plantago lanceolata (10.6 %), dog dander (10.4 %), and Pinus radiata (7.5 %). The sensitisation to indoor aeroallergens, was similar in all age groups and it was less important than that of pollens. The prevalence of sensitisation to grasses was the greatest in all ages and the house dust mites sensitization was the second most prevalent. The highest pollens counts in this region could explain the early sensitisation even in young children

Cova da Beira es una región interior del centro de Portugal con una población de 93.000 habitantes. Los primeros recuentos de polen realizados en Portugal revelaron que en esta zona se hallan los valores más elevados del país. El objetivo del presente estudio fue evaluar la sensibilización a los aeroalérgenos en una población alérgica según los grupos de edad. En un período de 5 años (1995-2000) se observaron 1.790 pacientes ambulatorios consecutivos con posibles síntomas alérgicos. En el estudio incluimos a los 557 pacientes pediátricos (de <= 15 años) observados (317 niños (57 %) y 240 niñas (43 %), con una edad media de 7,6 <= 4,2 años). Se dividieron en tres grupos de edad (grupo I: <= 5 años; grupo II: 6-10 años; grupo III: 11-15 años). Se sometieron 371 pacientes a pruebas cutáneas con aeroalérgenos. El 86,5 % de los mismos estaban sensibilizados como mínimo a un extracto de alergeno. Los alérgenos más significativos frente a los que se observó sensibilización fueron: mezcla de hierbas (44,9 %), D. pteronyssinus (32,5 %), D. farinae (29,1 %), Olea europea (27,5 %), Parietaria judaica (23,4 %), caspa de gato (16,1 %), Artemisia vulgaris (17,6 %), Robinia pseudoacacia (12,2 %), Platanus acerifolia (11,4 %), Tilia cordata (11,4 %), mezcla de mohos (11,2 %), Plantago lanceolata (10,6 %), caspa de perro (10,4 %) y Pinus radiata (7,5 %). La sensibilización a los aeroalérgenos domésticos fue similar en todos los grupos de edad y menos importante que la de los pólenes. La prevalencia de la sensibilización a las hierbas fue la principal en todos los grupos de edad, siendo la segunda en prevalencia la sensibilización a los ácaros domésticos. Los elevados niveles de polen de la región podrían explicar la sensibilización precoz incluso en niños de muy corta edad

Safety and tolerability of ultra-Rush (20 minutes) sublingual immunotherapy in patients with allergic rhinitis and/or asthma

Background: The safety and good tolerability of sublingual immunotherapy (SLIT) has already been proved in allergic patients, but only one study has investigated the occurrence of immediate adverse reactions in allergic patients after a 2-hour ultra-rush regimen of SLIT performed with a chemically modified extract (sublingual monomeric allergoid, Lais®, Lofarma S.p.A., Milan). The objective of the present study was to evaluate the occurrence of immediate adverse reactions in allergic patients after a very fast (20 minutes) ultra-rush regimen of sublingual allergoid SLIT. Methods and results: We studied 105 patients: 28 children (20 male, mean age 13.3 ± 2.1 yr) and 77 adults (29 male, mean age 34.7 ± 9.9 years) with a history of intermittent/persistent rhinitis or intermittent/mild persistent asthma due to House Dust Mite (n = 56), Parietaria (n = 34) and Timothy-grass (n = 15) The build-up ultra-rush phase involved the administration, every five minutes, of increasing doses of the sublingual allergoid SLIT. All patients tolerated the treatment very well. Only one patient out of 105 (0.9 %) had a mild local symptom (gastric pirosis) that occurred 30 minutes after the last initial dose and spontaneously disappeared as the treatment was continued. Conclusions: These data show the excellent safety and tolerability profile of an ultra-rush SLIT regimen performed with a chemically modified extract, even when high doses were administered through an extremely short induction phase (20 minutes), thus confirming the previously reported resultsBackground: The safety and good tolerability of sublingual immunotherapy (SLIT) has already been proved in allergic patients, but only one study has investigated the occurrence of immediate adverse reactions in allergic patients after a 2-hour ultra-rush regimen of SLIT performed with a chemically modified extract (sublingual monomeric allergoid, Lais®, Lofarma S.p.A., Milan). The objective of the present study was to evaluate the occurrence of immediate adverse reactions in allergic patients after a very fast (20 minutes) ultra-rush regimen of sublingual allergoid SLIT. Methods and results: We studied 105 patients: 28 children (20 male, mean age 13.3 ± 2.1 yr) and 77 adults (29 male, mean age 34.7 ± 9.9 years) with a history of intermittent/persistent rhinitis or intermittent/mild persistent asthma due to House Dust Mite (n = 56), Parietaria (n = 34) and Timothy-grass (n = 15) The build-up ultra-rush phase involved the administration, every five minutes, of increasing doses of the sublingual allergoid SLIT. All patients tolerated the treatment very well. Only one patient out of 105 (0.9 %) had a mild local symptoms (gastric pirosis) that occurred 30 minutes after the last initial dose and spontaneously disappeared as the treatment was continued. Conclusions: These data show the excellent safety and tolerability profile of an ultra-rush SLIT regimen performed with a chemically modified extract, even when high doses were administered through an extremely short induction phase (20 minutes), thus confirming the previously reported results

Antecedentes: La seguridad y la buena tolerancia de la inmunoterapia por via sublingual (ITSL) se ha comprobado en pacientes alérgicos, pero solamente en un estudio se ha investigado la aparición de reacciones adversas immediatas en pacientes alérgicos con un protocolo con inmunoterapia sublingual con dosificación ultra rápida en dos horas, administrando un extracto químicamente modificado (alergoide monomérico sublingual, Lais®, Lofarma S.p.A., Milan). El objetivo del presente estudio fue evaluar la aparición de reacciones adversas inmediatas en pacientes alérgicos con un protocolo de dosificación ultra rápido (20 minutos) de un alergoide sublingual (ITSL). Métodos y resultados: Estudiamos 105 pacientes: 28 niños (20 varones, promedio de edad 13,3 ± 2,1 años) y 77 adultos (29 varones, promedio de edad 34,7 ± 9,9 años) con historia clínica de rinitis intermitente/persistente o asma intermitente/leve persistente causada por ácaros del polvo (n = 56), Parietaria (n = 34) y fleo de los prados (Timothy grass) (n = 15). La fase de concentración ultra rápida consistió en la administración por via sublingual de dosis crecientes del alergoide, cada cinco minutos. Todos los pacientes toleraron el tratamiento satisfactoriamente. Sólo un paciente entre los 105 (0,9 %) presentó síntomas locales leves (pirosis gástrica) durante los 30 minutos posteriores a la última dosis inicial que remitieron cuando se discontinuó el tratamiento. Conclusiones: Estos datos demuestran el excelente nivel de seguridad y tolerancia de un régimen de dosificación ultra rápido de ITSL con un extracto químicamente modificado, aún con la administración de dosis elevadas durante una fase de inducción extremamente corta (20 minutos), lo cual confirma los resultados anteriormente presentados

Specific immune response to Parietaria judaica plant profilin: a low T cell proliferative response supports high IgE and skin prick test

Background: allergic disease caused by Parietaria judaica (Pj) has been widely documented in Mediterranean area. Profilins have been identified as widely distributed allergenic proteins. The role of Pj profilin in specific immune response in Pj-sensitized patients is unknown. Methods: skin prick test and determination of specific and total IgE levels in serum were performed in all patients (n = 28) and non-allergic controls (n = 18). Peripheral blood mononuclear cells (PBMC) were isolated from both groups and stimulated with crude extract or highly purified Pj profilin. The production of type I and type II cytokines was determined by specific and polyclonal stimuli in patients and controls. T-cell lines specific to Pj profilin were established and cross-reactivity with another highly purified profilin from Phleum pratense (Phl p) was evaluated. Results: Pj profilin-sensitized patients showed a small but significantly increased in T-cell proliferative response to this profilin compared with non-atopic controls. The production of interleukin (IL)-4 and interferon (IFN)-γ in response to the specific stimulus was undetectable. However, the production of IL-4 in response to a polyclonal stimulus [phytohemagglutinin (PHA)] was significantly higher in atopic patients than in controls. The T-cell response did not correlate with the magnitude of response to skin prick tests with Pj profilin or with Pj-specific serum IgE levels. In addition, the production of IL-4 in response to a polyclonal stimulus (PHA) did not correlate with the individual skin prick tests to Pj profilin or with Pj-specific IgE levels in serum. The T-cell lines tested showed no cross-reactivity with Phl p profilin. Conclusions: our results suggest that Pj profilin is partly responsible for the T-cell-mediated response in patients allergic to Pj. The high skin reactivity to Pj profilin is these patients was accompanied by a small increase in the T-cell response to this profilin. The response was highly specific since Pj profilin specific T-cell lines showed no cross-reactivity with a highly homologous profilin from Phl p. The lack of correlation between the proliferative T-cell response and polyclonal IL-4 production with allergen-specific serum IgE and skin reactivity probably indicates that some of the responding T-cells may be involved in immune reactions other than those supporting IgE production (AU)

Antecedentes: La enfermedad alérgica originada por Parietaria judaica (Pj) ha sido ampliamente documentada en la zona Mediterránea. Las profilinas han sido identificadas como proteínas alergénicas de amplia distribución. El papel de la profilina de Pj en la respuesta inmunológica específica en pacientes sensibilizados a la Pj es desconocida. Métodos: Se determinaron en todos los pacientes (n = 28) y controles normales no atópicos (n = 18) el test de reactividad cutánea y los niveles tanto de IgE específicas como totales en suero. Se aislaron células mononucleares de sangre periféica (PBMC) en ambos grupos y fueron estimuladas con el extracto crudo o con la profilina altamente purificada de Pj. Se determinaron también la producción de citocinas de tipo I y de tipo II, con estimulaciones específicas o policlonales en pacientes y controles. Establecimos líneas de células T específicas por la profilina de Pj y evaluamos la reactividad cruzada con otra profilina altamente purificada de Phleum pratense (Phl p). Resultados: Los pacientes sensibilizados a la profilina de Pj muestran un ligero pero significativo incremento en la respuesta proliferativa T frente la profilina respecto con el grupo de controles no alérgicos. La producción de IL-4 y IFN-γ en respuesta a estímulos específicos fue indetectable. Sin embargo, la producción de IL-4 en respuesta a un estímulo policlonal (PHA), fue significativamente mayor en pacientes atópicos que en los controles no alérgicos. La respuesta celular T no se corresponde ni con la magnitud de la repuesta en la reactividad cutánea a la profilina de Pj ni con los niveles de IgE en suero específicos para la Pj. La producción de IL-4 frente a un estímulo policlonals (PHA), no se correlaciona con la reactividade cutánea a la profilina ni con los niveles de IgE en suero. Las líneas específicas T testadas no mostraron ninguna reactividad cruzada con la profilina de (Phl p). Conclusiones: Nuestros resultados sugieren que la profilina de Pj es responsable en parte de la respuesta mediada por células T en pacientes alérgicos a la Pj. La elevada reactividad cutánea de estos pacientes, se acompaña de un pequeño incremento en la respuesta de células T a esta profilina. La respuesta es muy específica porque las líneas de células T no presentaron respuesta cruzada con la profilina altamente homóloga de Phl p. La falta de correlación entre la respuesta proliferativa de las células T y la producción policlonal de IL-4 con los niveles de IgE específicos en suero y la reactividad cutánea probablemente indica que algunas de las células T respondedoras estarían involucradas en una reacción inmunitaria diferente a la que promueve la producción de IgE (AU)