Sal k 5, a member of the widespread Ole e 1-like protein family, is a new allergen of Russian thistle (Salsola kali) pollen.

BACKGROUND: Salsola kali is an Amaranthaceae weed with important repercussions for pollinosis in temperate areas. Ole e 1-like members are relevant allergens in pollen from different species. We aimed to characterize and produce as recombinant allergen S. kali Ole e 1-like protein. METHODS: The natural allergen was purified at homogeneity after three chromatographic steps. Specific cDNA was sequenced and expressed in Pichia pastoris yeast. Structural relationships of natural and recombinant forms were carried out by 2D electrophoresis and spectroscopic analyses. Its immunological relevance was analyzed by ELISA and immunoblotting using an IgG antiserum and monoclonal antibodies specific to Ole e 1, as well as sera from 57 allergic patients recruited from two Spanish regions where this pollinosis is frequent. RESULTS: The purified allergen, Sal k 5, is an acidic glycoprotein of 151 amino acid residues and 17,628 Da of molecular mass. Its amino acid sequence exhibits 68 and 32% identity with the allergens of Che a 1 and Ole e 1, respectively. The recombinant protein was correctly processed and its structural and immunologic equivalence to the natural form was proven. A sensitization frequency between 30 and 40% was observed in pollinic patients from the center and east coast of Spain. CONCLUSIONS: Sal k 5 is a member of the Ole e 1-like protein family which can be considered an important allergen from S. kali. Its inclusion in diagnosis protocols would allow the accurate defining of patients allergic to this pollen.

A pectin methylesterase as an allergenic marker for the sensitization to Russian thistle (Salsola kali) pollen.

BACKGROUND: Chenopodiaceae pollen is considered the main cause of pollen allergy in desert countries and its incidence is world-wide increasing by the desertization of extensive zones. Although the correlation between the sensitization to Chenopodium album and Salsola kali pollens of patients suffering from allergy to Chenopodiaceae pollens is high, a significant number of patients exhibited IgE sensitivity exclusively towards S. kali. OBJECTIVE: To analyse this differential reactivity and to purify, clone and characterize the putative responsible allergen. METHODS: Immunoblotting was used to analyse the IgE binding to pollen extract for S. kali and C. album. The protein was isolated by two chromatographic steps and characterized by Edman degradation, mass spectrometry, finger print analysis and Concanavalin A lectin staining. Specific cDNA was amplified by polymerase chain reaction, cloned in Escherichia coli and sequenced. Immunologic characterization was performed by immunoblotting, enzyme-linked immunoassay detection and inhibition experiments using sera from 11 patients allergic to S. kali pollen. RESULTS: cDNA codifies for a mature protein of 339 amino acids plus a putative signal peptide of 23 residues and it belongs to the plant pectin methylesterase (PME) family. It is a mildly basic and polymorphic protein and was recognized by the IgE from all the patients allergic to S. kali included in the study, and was called Sal k 1. The protein was not recognized in the C. album pollen extract using the sera of these patients. CONCLUSION: Sal k 1 is a protein from the PME family with a high allergenic relevance. Considering this allergen as responsible for the different sensitization between S. kali and C. album pollen, it may be a useful marker to classify patients allergic to Chenopodiaceae allowing a safer and more specific immunotherapy.

Clinical efficacy and safety of a depigmented and glutaraldehyde polymerized therapeutic vaccine of Parietaria judaica.

BACKGROUND: The inhalation of Parietaria judaica pollen is a common cause of allergic respiratory diseases in the Mediterranean area. The objective of this study was to investigate the safety and clinical efficacy of a chemically modified (depigmented and glutaraldehyde-polymerized) vaccine of Parietaria judaica. METHODS AND RESULTS: Thirty patients with a well-documented clinical history of seasonal rhinitis and clinical sensitivity to Parietaria judaica pollen were included in a randomized trial during 12 months. The study was conducted following good clinical practices and appropriate consent forms were signed. Patients were divided into 2 groups of 15 individuals; group A received the modified extract and group C did not receive specific immunotherapy. Any adverse event was recorded to assess safety. Symptom scores, symptomatic medication use and the results of specific nasal challenges (before and after 12 months of treatment) were recorded to evaluate clinical efficacy. The treatment schedule consisted of an incremental phase of 5 injections and a maintenance dosage of 0.5 ml per month. Each patient received 14 injections during this period. All the patients completed the trial and no adverse reactions related to immunotherapy were recorded. A significant difference (p < 0.001) in symptom scores and overall use of symptomatic medication was observed between the two groups, being both scores lower in group A. No significant differences in nasal sensitivity existed before treatment among the 2 groups. However, after 12 months, a significant difference (p < 0.05) was observed only in group A patients, who showed a significant improvement in specific nasal challenges. CONCLUSIONS: Immunotherapy with depigmented and glutaraldehyde-polymerized extract of Parietaria judaica pollen is safe and effective to treat patients with allergic rhinitis and clinical sensitivity to this pollen.

Clinical efficacy and safety of a depigmented and glutaraldehyde polymerized therapeutic vaccine of Parietaria judaica

Background: The inhalation of Parietaria judaica pollen is a common cause of allergic respiratory diseases in the Mediterranean area. The objective of this study was to investigate the safety and clinical efficacy of a chemically modified (depigmented and glutaraldehyde-polymerized) vaccine of Parietaria judaica. Methods and results: Thirty patients with a well-documented clinical history of seasonal rhinitis and clinical sensitivity to Parietaria judaica pollen were included in a randomized trial during 12 months. The study was conducted following good clinical practices and appropriate consent forms were signed. Patients were divided into 2 groups of 15 individuals; group A received the modified extract and group C did not receive specific immunotherapy. Any adverse event was recorded to assess safety. Symptom scores, symptomatic medication use and the results of specific nasal challenges (before and after 12 months of treatment) were recorded to evaluate clinical efficacy. The treatment schedule consisted of an incremental phase of 5 injections and a maintenance dosage of 0.5 ml per month. Each patient received 14 injections during this period. All the patients completed the trial and no adverse reactions related to immunotherapy were recorded. A significant difference (p < 0.001) in symptom scores and overall use of symptomatic medication was observed between the two groups, being both scores lower in group A. No significant differences in nasal sensitivity existed before treatment among the 2 groups. However, after 12 months, a significant difference (p < 0.05) was observed only in group A patients, who showed a significant improvement in specific nasal challenges. Conclusions: Immunotherapy with depigmented and glutaraldehyde-polymerized extract of Parietaria judaica pollen is safe and effective to treat patients with allergic rhinitis and clinical sensitivity to this pollen (AU)

Antecedentes: La inhalación del polen de Parietaria judaica es una causa frecuente de enfermedades respiratorias alérgicas en la región mediterránea. El objetivo de este estudio era investigar la seguridad y la eficacia clínica de una vacuna químicamente modificada (despigmentada y polimerizada con glutaraldehído) de Parietaria judaica. Métodos y resultados: Se incluyó en un estudio aleatorizado de 12 meses de duración a 30 pacientes con historia clínica bien documentada de rinitis estacional y sensibilidad clínica al polen de Parietaria judaica. El estudio se llevó a cabo conforme a las buenas prácticas clínicas y se firmaron los formularios de consentimiento apropiados. Se distribuyó a los pacientes en dos grupos de 15 sujetos; el grupo A recibió el extracto modificaco y el grupo C no recibió inmunoterapia específica. Para evaluar la inocuidad se registraron las reacciones adversas. Para evaluar la eficacia clínica se registraron las puntuaciones de los síntomas, el uso de medicación sintomática y los resultados de pruebas de provocación nasales específicas (antes y después de 12 meses de tratamiento).El régimen de tratamiento consistió en una fase de incremento de 5 inyecciones y una posología de mantenimiento de 0,5 ml al mes. Cada paciente recibió 14 inyecciones durante ese período. Todos los pacientes se sometieron al ensayo completo y no se registraron reacciones adversas relacionadas con la inmunoterapia. Se observó una diferencia significativa (p < 0,001) en las puntuaciones de los síntomas y el uso global de medicación sintomática entre los dos grupos; ambas puntuaciones fueron menores en el grupo A. Antes del tratamiento no se observaron diferencias significativas en la sensibilidad nasal de los dos grupos. Sin embargo, al cabo de 12 meses, se observó una diferencia considerable (p < 0,05) sólo en los pacientes del grupo A, los cuales experimentaron una mejoría significativa en pruebas de provocación nasales específicas. Conclusiones: La inmunoterapia con extracto despigmentado y polimerizado con glutaral de hído de polen de Parietaria judaica es segura y eficaz para tratar a los pacientes con rinitis alérgica y sensibilidad clínica a este polen (AU)

Significant improvement in symptoms, skin test, and specific bronchial reactivity after 6 months of treatment with a depigmented, polymerized extract of Dermatophagoides pteronyssinus and D. farinae.

BACKGROUND: The efficacy of allergen immunotherapy using depigmented and polymerized extracts has been previously shown for Olea europaea, Phleum pratense, and Parietaria judaica. The objective of this study was to evaluate the efficacy after 3 and 6 months of treatment of a depigmented, polymerized extract of a mixture of Dermatophagoides pteronyssinus and D. farinae. METHODS: A group of 22 patients suffering from asthma and monosensitized to mites was treated with the mixture of modified allergen extract. A group of 11 mite-sensitive, asthmatic patients receiving only pharmacological treatment was used as control. The study was open, parallel, controlled, and random-allocated. Objective and subjective criteria, such as changes in D. pteronyssinus-specific bronchial hyperreactivity, visual scale, and medication/symptom scores were used to evaluate efficacy. Each patient received a built-up phase of 6 injections in 5 weeks, followed by 5 injections of the maintenance dose, which consisted of 42.5 micrograms of depigmented, polymerized extract of D. pteronyssinus and 32.5 of D. farinae. The Friedman test was used to compare the results of the specific bronchial challenges at baseline and after 3 and 6 months of treatment. RESULTS: A significant difference in the amount of native extract of D. pteronyssinus needed to produce a drop of 20% in the FEV1 (p = 0.0029) in the immunotherapy-treated group was found. In this group, the median allergen potency needed at baseline was 0.6 HEP (35 micrograms) vs. 3.96 HEP (232 micrograms) at the end of the study, whereas no difference (median 0.6 vs. 0.57 HEP) was found in the control group. At the end of the study, 10 patients in the immunotherapy treated group vs. 1 in the control group needed more than twice the amount of allergen than at baseline to experience a 20% drop in FEV1 (p = 0.03). Symptom and medication scores and visual scale evaluation did also show a significant improvement after 3 and 6 months of treatment only in active group. A significant decrease in skin test reactivity was also detected in the active group after 6 months, which needed a median of 3 times more allergen to elicit the same reaction as histamine (10 HEP) (p = 0.028), whereas no changes were found in control group. No serious side effects were registered. CONCLUSIONS: Depigmented polymerized extracts of D. pteronyssinus and D. farinae are safe and effective in the treatment of mite allergic asthmatic patients, and provide clinical benefit in the shock organ after 6 months of treatment. Skin test reactivity, symptom and medication scores were also improved. Depigmented polymerized extracts of D. pteronyssinus and D. farinae induce clinical protection against a native extract as verified by specific bronchial challenges.

Aerobiology of Artemisia airborne pollen in Murcia (SE Spain) and its relationship with weather variables: annual and intradiurnal variations for three different species. Wind vectors as a tool in determining pollen origin.

Detailed results from a 2-year survey of airborne pollen concentrations of Artemisia in Murcia are presented. Three consecutive pollen seasons of Artemisia occurring each year, related to three different species (A. campestris, A. herba-alba and A. barrelieri), were observed. A winter blooming of Artemisia could explain the incidence of subsequent pollinosis in the Murcia area. With regard to meteorological parameters, mathematical analyses showed relationships between daily pollen concentrations of Artemisia in summer-autumn and precipitations that occurred 6-8 weeks before. The cumulative percentage of insolation from 1 March seemed to be related to blooming onsets. Once pollination has begun, meteorological factors do not seem to influence pollen concentrations significantly. Intradiurnal patterns of pollen concentrations were similar for late summer and winter species (A. campestris and A. barrelieri). During autumn blooming (A. herba-alba), the intradiurnal pattern was particularly erratic. Theoretical values of wind run were obtained for each pollen season by the graphical sum of hourly wind vectors. When theoretical wind run was mapped onto the vegetation pattern, supposed pollen source locations were obtained for each hour. By comparing supposed hourly pollen origins with the intradiurnal patterns of pollen concentrations, it can be seen that this simple model explains variations in mean pollen concentrations throughout the day.

Incidence of Alternaria spores in the atmosphere of Murcia (SE Spain). Seasonal, monthly and intradiurnal variations.

Variation in Alternaria spores in the atmosphere of Murcia (SE Spain) were studied for 2 years. Using a Burkard volumetric sampler located about 19 m above ground level, spores were collected and counted hourly by light microscopy. The results suggest that species of Alternaria are present in the atmosphere of Murcia every day throughout the year. Mean daily concentration of Alternaria was 25 spores/m3. Seasonal variations showed the lowest concentrations during winter and the highest in autumn. In the monthly pattern two peaks were observed every year: the first in about May-June and the second occurring in October. In the intradiurnal variations, maximum concentrations were observed between 5:00-6:00 p.m. (Spanish official time). The variation of Alternaria spores in the atmosphere in Murcia seems to be related to different factors throughout the year.

[Incidence of adverse reactions to additives. Our experience over 10 years]. / Incidencias de reacciones adversas con aditivos. Nuestra experiencia de 10 años.

No published information exists about the incidence of food additives reactions in the general population. Most studies have been made in patients with urticaria and bronchial asthma. The majority of them lack an adequate design and, therefore, the reported results should be interpreted with extreme caution. In this article, we expose our ten years experience in this field. We have added up 1941 oral provocation tests, with an ample battery of additives, administering the tested substances directly or in aqueous or acid solution (1110 in patients with urticaria, mainly chronic urticaria, and 831 in asthmatic subjects, with or without aspirin intolerance). From these exhaustive data, we get the following conclusions: 1) in contrast with other-investigators, and using similar or even higher provocation doses, we get a very low incidence of adverse reactions. 2) We are sceptical that food additives play any role in chronic urticaria or in other cutaneous processes (only 0.63% provocation tests resulted in an urticarial exacerbation, and none of them was repeated after re-provocation). 3) In asthmatic patients, similar results were obtained, except with sulfites in acid solution challenge test (10% asthmatic exacerbations), possibly as a sign of nonspecific bronchial hyperreactivity. 4) The prescription of food additives free restrictive diets does not seem to be justified. The should be followed only by those patients with clear evidence of additives reactions. 5) In most cases, with punctual exceptions, the study of food additives reactions, in clinical allergy, implies a waste of time.

Allergenic cross-reactivity among pollens of Urticaceae.

Common antigenic determinants have been observed between Parietaria and Urtica dioica pollen. The four Parietaria pollens selected (P. judaica, P. officinalis, P. lusitanica and P. mauritanica) are shown to possess a high allergenic homology. IgE-binding structures, homologous to the P. judaica main allergenic polypeptide (Pj10), were found in the other species by immunodetection. Monoclonal antibodies specific to the Pj10 polypeptide recognized proteins from the four Parietaria pollens. Skin prick test and RAST inhibition yielded results that also indicated a high allergenic cross-reactivity among these pollens, with homologous peptides bearing common antigenic and allergenic determinants. On the other hand, U. dioica pollen showed only a slight allergenic similarity to Parietaria. The potential allergenic activity of these pollens is discussed.