Allergen Profile of London Plane Tree Pollen: Clinical and Molecular Pattern in Central Spain.

BACKGROUND AND OBJECTIVES: Platanus acerifolia (London plane tree) is a deciduous tree of the Platanaceae family. Sensitization to this plant varies with geography. Madrid, located in central Spain, has one of the highest London plane tree pollen concentration levels on the Iberian Peninsula. We evaluated both the clinical characteristics and the molecular sensitization pattern of patients with allergy to London plane tree pollen in the region of Madrid. METHODS: Thirty-eight patients allergic to London plane tree pollen were selected according to their clinical symptoms and positive results in skin prick testing and/or specific IgE determination. Serum was collected, and allergen components were evaluated using immunodetection techniques as well as ImmunoCAP. The IgE-binding proteins detected were identified and characterized using mass spectrometry. RESULTS: Analysis of serum samples from allergic patients revealed 9 IgE-binding bands in London plane tree pollen extract. Among these, the 45-kDa protein, which corresponded to Pla a 2, was detected in 76.3% of patients. However, the 18-kDa (Pla a 1) and 9-kDa (Pla a 3) bands were detected in 44.7% and 23.7% of sera, respectively. These results were confirmed using purified proteins. Characterization of the allergen revealed the 27-kDa protein to be glutathione-S-transferase. CONCLUSIONS: The molecular profile of patients sensitized to London plane tree pollen differs from that reported in studies from other locations. In the population we studied, the prevalence of Pla a 2 was higher than that of Pla a 1 and Pla a 3. In addition, the minor allergen previously referred to as Pla a 4 was characterized as glutathione-S-transferase.

Association Between the Seed Storage Proteins 2S Albumin and 11S Globulin and Severe Allergic Reaction After Flaxseed Intake.

BACKGROUND: Given the increased popularity of flaxseed in meals, several cases of allergy to these seeds have been reported. Little is known about the allergens implicated in hypersensitivity reactions to flaxseed. The present study aimed to identify the allergens involved in IgE-mediated reactions in 5 patients with a clinical history of severe systemic symptoms after flaxseed consumption. METHODS: Proteins that were potential allergens with IgE-binding capacity were purified from flaxseed extract using chromatography and identified via MALDI-TOF mass spectrometry. Immunoassays were performed using the 5 allergic patients' sera tested individually and as a pool. RESULTS: Immunoblotting of the flaxseed extract revealed a low-molecular-mass protein (around 13 kDa) in 4 of the 5 patients, while a protein of approximately 55 kDa was detected in 2 patients. The proteins were identified by mass spectrometry as flaxseed 2S albumin, which is included in the WHO/IUIS allergen nomenclature as Lin u 1, and 11S globulin. Inhibition assays revealed in vitro IgE-mediated cross-reactivity between Lin u 1 and peanut and cashew nut proteins, while IgE-mediated recognition of 11S globulin by patients' sera was partially inhibited by several plant-derived sources. CONCLUSIONS: Seed storage proteins from flaxseed were involved in the development of severe symptoms in the 5 patients studied and exhibited cross-reactivity with other allergenic sources. Besides the severity of flaxseed allergy in patients sensitized to 2S albumin, this is the first time that 11S globulin has been identified as a potential allergen. Taking these data into account should ensure a more accurate diagnosis.

Lettuce allergy is a lipid transfer syndrome-related food allergy with a high risk of severe reactions

Background and Objectives: Lipid transfer protein (LTP) sensitization is the most common cause of food allergy in the Mediterranean area, with peach allergy acting as the primary sensitizer in most cases. Lettuce has been described as a common offending food in patients with LTP syndrome. The aim of the study was to investigate the frequency and clinical expression of LTP syndrome in a sample of lettuceallergic patients. Methods: We determined specific IgE to Pru p 3 and lettuce in a sample of 30 patients with a diagnosis of lettuce allergy. Symptoms elicited by other LTP-containing plant-derived foods and the presence of cofactors were assessed. Results: The clinical symptoms of lettuce allergy were frequently severe, with 18 of the 30 patients experiencing anaphylaxis. All the patients had allergic reactions to other plant foods. Cofactors were involved in the clinical reactions of 13 of the 30 patients. Sensitization to pollens was found in 90% of patients. Conclusions: Lettuce allergy is found not as an isolated condition but in the context of LTP syndrome and it is characterized by severe reactions and frequent cofactor association (AU)

Introducción y Objetivo: La sensibilización a la LTP es la causa más frecuente de alergia alimentaria en el área mediterránea, siendo la alergia al melocotón el sensibilizador primario en la mayoría de los casos. La alergia a la lechuga ha sido descrita como un manifestación frecuente en los pacientes que sufren síndrome de LTP. El objetivo del estudio fue investigar la frecuencia del síndrome de LTP en una muestra de pacientes alérgicos a lechuga y evaluar su patrón clínico. Métodos: Se determinó la IgE específica a Pru p 3 y a lechuga en una muestra de 30 pacientes con un diagnóstico de alergia a la lechuga. Se evaluaron los síntomas con otras LTPs de alimentos de origen vegetal y la presencia de cofactores. Resultados: Los síntomas clínicos de la alergia a lechuga fueron frecuentemente graves, ya que 18/30 pacientes experimentaron anafilaxia. Todos los pacientes experimentaron reacciones alérgicas a otros alimentos vegetales. En 13/30 pacientes, los cofactores estaban implicados en las manifestaciones clínicas. Se observó que el 90% de los pacientes estaban sensibilizados a pólenes. Conclusiones: La alergia a la lechuga más que de forma aislada, ocurre en el contexto del síndrome LTP y se caracteriza por su frecuente asociación a cofactores y la gravedad de sus reacciones (AU)

ETP-46321, a dual p110α/δ class IA phosphoinositide 3-kinase inhibitor modulates T lymphocyte activation and collagen-induced arthritis.

Class IA phosphoinositide 3-kinases (PI3Ks) are essential to function of normal and tumor cells, and to modulate immune responses. T lymphocytes express high levels of p110α and p110δ class IA PI3K. Whereas the functioning of PI3K p110δ in immune and autoimmune reactions is well established, the role of p110α is less well understood. Here, a novel dual p110α/δ inhibitor (ETP-46321) and highly specific p110α (A66) or p110δ (IC87114) inhibitors have been compared concerning T cell activation in vitro, as well as the effect on responses to protein antigen and collagen-induced arthritis in vivo. In vitro activation of naive CD4(+) T lymphocytes by anti-CD3 and anti-CD28 was inhibited more effectively by the p110δ inhibitor than by the p110α inhibitor as measured by cytokine secretion (IL-2, IL-10, and IFN-γ), T-bet expression and NFAT activation. In activated CD4(+) T cells re-stimulated through CD3 and ICOS, IC87114 inhibited Akt and Erk activation, and the secretion of IL-2, IL-4, IL-17A, and IFN-γ better than A66. The p110α/δ inhibitor ETP-46321, or p110α plus p110δ inhibitors also inhibited IL-21 secretion by differentiated CD4(+) T follicular (Tfh) or IL-17-producing (Th17) helper cells. In vivo, therapeutic administration of ETP-46321 significantly inhibited responses to protein antigen as well as collagen-induced arthritis, as measured by antigen-specific antibody responses, secretion of IL-10, IL-17A or IFN-γ, or clinical symptoms. Hence, p110α as well as p110δ Class IA PI3Ks are important to immune regulation; inhibition of both subunits may be an effective therapeutic approach in inflammatory autoimmune diseases like rheumatoid arthritis.

[BIOBADADERM, the Spanish Registry of Adverse Events Associated with Biologic Drugs in Dermatology: first report]. / BIOBADADERM: registro español de acontecimientos adversos de terapias biológicas en Dermatología. Primer informe.

BACKGROUND AND OBJECTIVES: The Working Group on Psoriasis of the Spanish Academy of Dermatology and Venereology has initiated BIOBADADERM, a registry of patients with psoriasis receiving treatment with biologic drugs, in order to assess the long-term risk of adverse events (AEs). MATERIAL AND METHODS: A multicenter study was undertaken in 2 cohorts of patients with psoriasis: patients receiving biologic therapy and patients receiving nonbiologic systemic therapy other than phototherapy. Similar numbers of patients were included in each group. Information was recorded on demographic and clinical variables, treatment, and relevant AEs. The risk of specific AEs was determined by comparison of the frequencies for those events in the 2 cohorts. RESULTS: Data on the 2 cohorts were evaluated for the period from October, 2008 to November, 2009 alongside retrospective data on patients treated with biologics since 2005. Thirteen Spanish hospitals participated in the study. A total of 632 patients were included in the analysis: 417 treated with biologic drugs and 215 controls. Suspension of biologic therapy due to AEs was rare (72 cycles, 10%). A total of 232 AEs were reported in patients receiving biologic therapy. The majority were not serious. The most frequent AEs were infections (mostly upper respiratory tract infections and nasopharyngitis), followed by conditions affecting the skin or subcutaneous tissue. Forty-three AEs were reported in control subjects. The most frequent events were metabolic and nutritional abnormalities and abnormal transaminase levels. Comparison of the incidence of any AE in patients treated with biologics compared with control subjects revealed a relative risk of 2.2 (P<.001) The relative risks of infections or infestations and disorders of the skin or subcutaneous tissue in patients receiving biologic drugs were 23 (P<.01) and 4.9 (P<.05), respectively. CONCLUSIONS: Patients treated with biologic drugs had a greater number of AEs, particularly infections and skin conditions. Definitive conclusions, however, are difficult to draw due to the small number of patients included in the registry, particularly in the control cohort, and the short follow-up period. Differences in the percentages of events reported by the different hospitals reveal the difficulties associated with the concept of AEs in clinical practice and highlight the need to harmonize criteria in the future. Since the problems identified in this analysis should be overcome in future years, we expect BIOBADADERM to become an important source of information on the safety profile of biologic drugs in dermatology.

Characterization of IgE epitopes of Cuc m 2, the major melon allergen, and their role in cross-reactivity with pollen profilins.

BACKGROUND: Plant profilins are described as minor allergens, although with some exceptions in foods such as melon, watermelon or orange. In fact, they could be responsible for many cross-reactions among distantly related species. This is likely to be a consequence of the presence of common epitopes. OBJECTIVE: To characterize the B epitopes of Cuc m 2, a model of plant food profilin, using phage display techniques and to compare with other profilins, such as those of timothy grass and birch pollen, and human I profilin, to understand the mechanism of cross-reaction among members of this family. METHODS: IgE of melon-allergic patients was used to select clones from a phage display 12 mer peptide library. After two rounds of screening, Cuc m 2-specific clones were eluted and the DNA insertion sequenced. The residues of each clone were mapped on the Cuc m 2 surface to define a mimotope, which was also localized on the three-dimensional surfaces of other profilins. RESULTS: Seventeen melon-allergic patients were selected. Sera from each of them recognized the melon profilin, Cuc m 2, but the majority also recognized Phl p 12 or Bet v 2, timothy grass-, and birch-pollen profilins, respectively. A Cuc m 2 mimotope was defined and mapped onto its surface giving the following sequence: S(2)W(3)A(5)Y(6)D(9)H(10)T(111)P(112)G(113)Q(114)N(116)M(117)R(121)L(122). The homologous residues in Phl p 12 and Bet v 2 had almost identical sequences. By contrast, the homologous sequence in human profilin showed many differences. CONCLUSIONS: The identified mimotope could be involved in cross-reactions among food and pollen profilins. Many of these cross-reactions observed in the clinical realm could be explained by the presence of a common epitope found in food and pollen allergens. A new strategy of immunotherapy based on this IgE region could be used in alternative immunotherapy strategies.

Immunotherapy with depigmented glutaraldehyde-polymerized extracts: changes in quality of life.

BACKGROUND: Immunotherapy (IT) with modified allergens reduces allergic rhinitis (AR) symptoms and medications requirements. Improvement of quality of life (QOL) is a key point in the treatment of AR. The aim of this study was to provide evidence of changes related to the patient's QOL (well-being) induced by a modified (depigmented glutaraldehyde-polymerized) therapeutic vaccine and of its safety. MATERIAL AND METHODS: Fifty-three patients with a well-documented clinical history of seasonal AR sensitized to Dactylis glomerata and Olea europaea pollens were included in a randomized clinical trial. Twenty-five patients (Group-A) received a mixture of D. glomerata and O. europaea pollen extracts and 28 patients received placebo (group-C). Any adverse event was recorded and graded in accordance with EAACI guidelines. RQLQ was recorded before the treatment (pollen season 2000) and after 1 year of treatment (pollen season 2001). Dose-response skin prick test with each allergen extract was conducted at baseline and at the end of the study. RESULTS: Each patient received 17 injections during this period. All patients completed the trial and no systemic adverse reactions were recorded. Symptom scores (P<0.001) and medication requirements (P<0.001) were significantly reduced in the IT group during the pollen season. This patient group also experienced greater and statistically significant improvement in overall RQLQ score and in five of the seven domains, all of them surpassing the threshold of 'minimal important difference' of 0.5 points. CONCLUSIONS: Results of this study provided evidence that IT with depigmented, glutaraldehyde-modified allergen extracts was well-tolerated and added beneficial effects to AR treatment in pollen allergic patients eliciting an improvement in QOL enough to justify a change in the patient's treatment.

Identification of Cucumisin (Cuc m 1), a subtilisin-like endopeptidase, as the major allergen of melon fruit.

BACKGROUND: Allergenic components in melon extracts have not been described in spite of the fact that melon (Cucumis melo) is a frequent allergy-eliciting fruit. The aim of this study was to evaluate allergenic components in melon extract and to report the identification of cucumisin as a major melon allergen. MATERIALS AND METHODS: Sera from 35 patients allergic to melon were selected on the basis of clinical symptoms, skin prick tests and oral challenge test. Allergenic components were detected by sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting. Molecular characterization of IgE-binding bands was performed by N-terminal amino acid sequencing. RESULTS: More than 10 IgE-binding bands, between 10 and 80 kDa, were identified in melon extract. Out of them, four IgE-binding bands were major allergens: 14 kDa, 36 kDa, 54 kDa and 67 kDa. These major allergens, except 14 kDa band, showed the same N-terminal sequence: T-T-R-S-W-D-F-L. Research conducted with protein databases identified this N-terminal sequence as cucumisin, an alkaline serine protease, which shares structural homology with microbial subtilisin. The molecular mass of the identified bands corresponds with different molecular forms of cucumisin produced during the processing or degradation of the enzyme: 67 kDa native cucumisin, 54 kDa mature cucumisin and 36 kDa NH2-terminal cucumisin fragment. CONCLUSION: Cucumisin (Cuc m 1) and several N-terminal cucumisin fragments are the major allergens of melon. The ubiquitous distribution of this protein family (cucumisin-like proteases) in many plant species and its high structural similarity suggest its potential role as a new panallergen in plant foods.

Pru p 3 (LTP) content in peach extracts.

BACKGROUND: Lipid transfer proteins are molecules widely distributed in fruits. Sensitization to LTP is frequent in fruit sensitive patients. The aims of this study were to purify LTP and to assess the content of LTP in ripe peach peel and pulp extracts by ELISA inhibition using polyclonal antibodies. METHODS: LTP was purified from ripe yellow peach peel by two different column chromatography methods. A polyclonal antibody was produced by injecting purified LTP into two New Zealand white rabbits. ELISA inhibition and rabbit monospecific polyclonal antibody were used to calculate the LTP content in Springcrest and Miraflores varieties of peach peel and pulp extracts. Purified LTP (2.5 mg/ml) was used to skin test 24 peach-sensitive patients. RESULTS: The purified LTP showed a single band at approximately 9 kDa. The polyclonal antibody raised anti LTP recognized only the LTP molecule in the peach extracts. LTP content, expressed in micro g/mg of freeze-dried extract in four extracts were: yellow peach peel, 15.48; yellow peach pulp 2.25; red peach peel 14.67 and red peach pulp 1.84. Twenty patients (83.3%) had a positive skin test with purified LTP. CONCLUSIONS: We have developed a system to determine the concentration of LTP in peach extracts. LTP in peel extracts is approximately seven times greater than in pulp.

Allergy to plant-derived fresh foods in a birch- and ragweed-free area.

BACKGROUND: Allergy to plant-derived fresh foods has often been reported in geographical areas where birch or ragweed pollens are frequent and has been attributed to cross-reactivity to pollens. OBJECTIVE: The aim of this study has been to evaluate allergy to plant-derived fresh foods among pollen-allergic patients from a birch and ragweed-free area. METHODS: Ninety-five pollen-allergic patients took part in the study. The study consisted of a questionnaire, skin prick tests and challenge tests. Pollen skin tests to five grasses, eight trees and seven weeds were performed in duplicate. Prick tests (prick by prick) and challenge tests were carried out with the fresh foods. RESULTS: Most patients allergic to pollens were sensitized to grass (Lolium and Phleum; 97.9%), followed by tree (Olea; 82.1%) and weed pollens (Plantago; 64.2%). 35 of the 95 pollen-allergic patients had positive skin test responses to some plant-derived fresh foods, the highest percentage corresponding to several fruits in the Rosaceae family (peach and pear, 26.3%), followed by Cucurbitacea fruits (melon, 13.7%). The 21. 05% of the pollen-allergic patients were allergic to some type of plant-derived fresh food. Peach was the plant-derived fresh food which most frequently elicited allergy symptoms (12.6%), followed by melon (7.36%). The cluster of positive responses to Rosaceae fruits was higher for skin testing than for challenge testing. CONCLUSION: Peach was the most important allergy provoking fruit in a birch and ragweed free-area where apples were consumed at a rate of two times more than peaches and the patients allergic to pollen were principally sensitized to grass pollens.

Cryptogenic gelastic epilepsy of frontal lobe origin: a paediatric case report.

Gelastic (laughing) seizures are an uncommon seizure type which in most cases has an organic cerebral pathology and specifically a hypothalamic hamartoma. The interictal EEG frequently shows focal activity. This report describes a 3 1/2-year-old boy who presented with episodes of unmotivated laughter associated with other epileptic symptomatology before the age of 3 years. Prolonged ambulatory EEG monitoring recorded electroclinical seizures starting in the right frontal area and spreading to the adjacent frontotemporal region. Neurological examination and brain magnetic resonance imaging were normal. Vigabatrin resulted in immediate remission of the seizures and normalization of the EEG.

Pollen allergy in peach-allergic patients: sensitization and cross-reactivity to taxonomically unrelated pollens.

BACKGROUND: Fruit allergy has been attributed to cross-reactive IgE to pollens and has been associated with a particular pollen sensitization. OBJECTIVE: The aim of the study was to evaluate sensitization to several taxonomically unrelated pollens in peach- and pollen-allergic patients and to study cross-reactivity between them. METHODS: One hundred sixty-five patients were evaluated: 70 peach- allergic patients together with 95 pollen-allergic patients (control group). Pollen skin tests in duplicate were performed to 5 grasses, 8 trees, and 7 weeds. Cross-reactivity between peach and taxonomically diverse pollens was determined by radioallergosorbent inhibition and Western blot inhibition tests. Experiments were also carried out after preadsorption of the sera with purified natural profilin. RESULTS: The skin test results revealed that peach-allergic patients frequently reacted to most pollens-grasses, weeds, and trees-even when some of these are not found in our geographic area. There was a statistically significant increase in sensitization frequency to most trees and weeds, with a statistically higher occurrence of asthma (odds ratio 2.98, 95% confidence interval 1.46-6.09). Inhibition test results provided evidence that taxonomically unrelated grasses, weeds, and trees produced various and substantial degrees of inhibition in specific IgE to peach and that the peach extract elicited strong inhibitions to those pollens. Profilin was found to be a relevant cross-reactive antigen in these patients. CONCLUSION: The results of this study provide evidence that peach allergy is linked to sensitization to several taxonomically unrelated pollens. This is attributable to the ubiquitous nature of the IgE binding determinants-such as profilins-between peach and taxonomically unrelated pollens.

A method for quantitation of food biologic activity: results with peach allergen extracts.

OBJECTIVE: The aim of this study was to develop a quantitative skin test assay for measurement of the biologic activity of food allergen extracts, as well as to report the results obtained with a peach extract labeled in food biological units (FBUs). METHODS: We prepared a biologically quantified peach extract. The biologic activity was measured in FBUs by assigning 100 FBU/ml to the biologic activity of the extract that elicited a wheal area with a geometric mean equal to that produced by the prick-prick method by using the food itself in a population of 30 patients allergic to food. We evaluated 265 patients, including 70 patients allergic to peach and 195 control subjects (100 nonatopic subjects and 95 subjects allergic to pollen). The biologically quantified peach extract was used during the study and was compared with four commercial peach extracts, which were labeled in weight per volume. RESULTS: The sensitivity of the nonstandardized commercial peach extracts varied from 4.3% to 74%, with biologic activity being very low in all of them. The sensitivity of the biologically quantified peach extract was 100%, with a 100% concordance between the prick-prick and the skin prick test results. CONCLUSION: These data demonstrate that if peach allergen preparations were biologically standardized in FBUs, the quality of these food extracts used for diagnosis would be improved.

Peach allergy pattern: experience in 70 patients.

We report the clinical characteristics of peach allergy encountered in a population of peach-allergic patients. We evaluated 165 patients. The 70 peach-allergic patients were diagnosed through clinical history, the skin prick-prick test, and open oral challenge and rub tests to peach. As a pollinic control group, 95 pollen-allergic patients were also evaluated. Some 49% of the patients were male and 51% female. The mean age was 20 +/- 8 years. Oral allergy syndrome (86%) was the most common symptom, followed by contact urticaria (61%) and systemic symptoms (26%). Some 67% of the patients were allergic to peach pulp and 36% reported symptoms related to canned peach. Canned peach and pulp symptoms were statistically associated (P < 0.01), and symptoms to canned peach were more frequently reported by patients with systemic symptoms (P < 0.05). On evaluation of the peach-allergic patients' characteristics, three risk factors--allergy to peach pulp, allergy to canned peach, and peach allergy in non-pollen-allergic patients--were found, indicating development of systemic symptoms on eating peach. Most of the peach-allergic patients (81%) also had pollen allergy, which was linked to a higher prevalence of asthma (73%) than in the pollen-allergic patients of the control group (48%); this difference was statistically significant (P < 0.01). Finally, two groups were clearly defined by the seriousness of the peach allergy--the non-pollen-allergic patients were more predisposed to the occurrence of systemic symptoms (> 50%), and the pollen-allergic patients to asthma (> 70%).