Oral myeloid cells uptake allergoids coupled to mannan driving Th1/Treg responses upon sublingual delivery in mice.

BACKGROUND: Polymerized allergoids coupled to nonoxidized mannan (PM-allergoids) may represent novel vaccines targeting dendritic cells (DCs). PM-allergoids are better captured by DCs than native allergens and favor Th1/Treg cell responses upon subcutaneous injection. Herein we have studied in mice the in vivo immunogenicity of PM-allergoids administered sublingually in comparison with native allergens. METHODS: Three immunization protocols (4-8 weeks long) were used in Balb/c mice. Serum antibody levels were tested by ELISA. Cell responses (proliferation, cytokines, and Tregs) were assayed by flow cytometry in spleen and lymph nodes (LNs). Allergen uptake was measured by flow cytometry in myeloid sublingual cells. RESULTS: A quick antibody response and higher IgG2a/IgE ratio were observed with PM-allergoids. Moreover, stronger specific proliferative responses were seen in both submandibular LNs and spleen cells assayed in vitro. This was accompanied by a higher IFNγ/IL-4 ratio with a quick IL-10 production by submandibular LN cells. An increase in CD4+ CD25high FOXP3+ Treg cells was detected in LNs and spleen of mice treated with PM-allergoids. These allergoids were better captured than native allergens by antigen-presenting (CD45+ MHC-II+ ) cells obtained from the sublingual mucosa, including DCs (CD11b+ ) and macrophages (CD64+ ). Importantly, all the differential effects induced by PM-allergoids were abolished when using oxidized instead of nonoxidized PM-allergoids. CONCLUSION: Our results demonstrate for the first time that PM-allergoids administered through the sublingual route promote the generation of Th1 and FOXP3+ Treg cells in a greater extent than native allergens by mechanisms that might well involve their better uptake by oral antigen-presenting cells.

Cluster immunotherapy with a glutaraldehyde-modified mixture of grasses results in an improvement in specific nasal provocation tests in less than 2.5 months of treatment.

BACKGROUND: Cluster immunotherapy is becoming increasingly used. It allows for a rapid build up phase and the administration of higher doses of allergen in a shorter period of time. OBJECTIVES: To evaluate the effect of short-term pre-seasonal immunotherapy using a glutaraldeyde-modified allergen vaccine in reducing specific nasal hyperreactivity in nasal challenge tests. MATERIALS AND METHODS: Thirty-three patients were selected. All patients had a positive history of allergic rhinitis and skin tests to grass pollen, although most of them (72.7%) were sensitized to other allergens as well. The study was conducted outside of the pollen season and the patients did not receive any pharmacological treatment during this period of time. Two randomized groups of patients were established; Group A: 22 patients (13 females and nine males) and Group B, 11 control patients (seven females and four males). Patients in Group A received immunotherapy with a vaccine containing 50% of the wild grasses Trisetum paniceum and Dactylis glomerata. All patients underwent titrated nasal provocation tests (NPT) before and after completion of the study (2.3 and 2.8 months for Groups A and B, respectively). The administration schedule consisted of 0.1 and 0.2 mL at day 1, followed by 0.3 and 0.5 mL at day 7, 0.5 mL after 2 weeks followed by 0.5 mL monthly. A single vial was used containing an allergen concentration of 10 000 TU/mL (105 microg of total protein and 24.6 microg of Group 1 plus 5 allergens/mL). A mean of 6.5 injections were administered to Group A patients between NPTs. RESULTS: There were no significant differences between both groups at the beginning of the study (P=0.48). At the end, only Group A patients needed significant greater threshold concentrations for a positive NPT than at the beginning (P=0.002). CONCLUSIONS: A short-term cluster pre-seasonal inmunotherapy with a modified vaccine containing a mixture of grass pollen is effective as determined by an objective measure after only a mean 2.3 months of treatment.

Trisetum paniceum (wild oats) pollen counts and aeroallergens in the ambient air of Madrid, Spain.

BACKGROUND: Madrid has a short but intensive grass pollen season, in which 79% of the total grass pollen load is released from the middle of May to the middle of June. The main objectives of this study were to quantify Trisetum paniceum (wild oats) aeroallergen in the atmosphere in Madrid from February to December 1996 and to correlate the aeroallergen concentrations with grass pollen counts. METHODS: Two different samplers were used to assess allergen exposure; a Burkard spore trap was used to collect pollen grains and a high-volume air sampler to collect airborne particles. A total of 182 air filters were collected and extracted in 1 ml of PBS and analysed by ELISA inhibition, using pooled sera from highly allergic individuals. RESULTS: T. paniceum aeroallergens were detected not only during the grass pollen season, but also before and after. Wild oat allergens had two main peaks of 1 and 1.9 microg/m(3), occurring in late May and July, respectively. The time series analysis established the existence of lags between the two main variables pollen counts and aeroallergen activity. Analysis of the data by the Spearman rank test and linear regression showed a weak correlation between grass allergenic activity and grass pollen counts (Spearman's rho = 0.29). Data obtained from time series analysis demonstrated that grass allergenic activity correlated strongly with current and 5-week-old grass pollen grain counts (r(2) = 0.73). CONCLUSIONS: Wild oats allergenic activity was detected during the entire year and not only during the pollen season. This fact is an important aspect to be considered in the clinical follow-up and treatment of grass pollen-sensitised patients in Madrid.

Platanus pollen as an important cause of pollinosis.

OBJECTIVE: The existence of Platanus pollinosis is not generally accepted despite the production of very large quantities of airborne Platanus pollen in many cities of the United States and Europe. The aim of this study was to investigate if Platanus pollen really contributes to the symptoms of the patients with pollinosis in the Madrid area. METHODS: We carried out systematic skin prick testing with Platanus pollen extract on 47 patients seen in our allergy center with spring-summer pollinosis symptoms. Each patient maintained symptom score diaries before, during, and after the Platanus pollination season. The average symptom scores were calculated and compared with the Platanus pollen counts. Measurements of specific IgE by ELISA and immunoblotting also were performed in each patient. RESULTS: The Platanus skin tests were positive in 33 of the 39 patients first seen with seasonal symptoms during Platanus pollen season and only in three of the eight patients without symptoms during Platanus exposure (Fisher's exact test; p < 0.05). Twenty-two of the 33 Platanus-positive skin test patients also had a positive ELISA result. Furthermore, the average 24-hour rhinitis symptom scores of the 39 patients first seen with seasonal symptoms during March through April showed significant correlation with Platanus pollen counts (r(s) = 0.57, p < 0.05). The immunoblot results suggest that a 17 kd pollen protein could be a major allergen in patients with Platanus pollinosis. CONCLUSIONS: Platanus pollen is an important cause of pollinosis in Madrid. A protein with a molecular weight of 17 kd appeared to be its major allergen.

Seasonal asthma caused by airborne Platanus pollen.

This work describes three patients suffering from bronchial asthma after being naturally exposed to airborne plane-tree pollen. The three patients gave immediate response in skin tests and dual response in bronchial provocations using Platanus hybrida extract. There was specific seric IgE activity against this/these antigen(s) with the CAP system. The three patients also showed significant correlation (P < 0.001) between their rhinitis and asthma symptom-scores registered on their diary cards and plane-tree pollen counts, collected using the Burkard spore trap. Among 187 patients living in Madrid and who came to our centre with a history of rhinitis and/or seasonal asthma, we found a prevalence of positive skin-prick tests to Platanus of 56%, only surpassed by gramineous pollen (Dactylis glomerata and/or Trisetum paniceum) 92% and Olea europaea 63%. The aerobiological sampling of the pollen content of the air in Madrid, carried out between 1 January 1979 and 31 December 1993 revealed an airborne presence (per cent of total yearly pollen count, mean of 1979-1993) of 14.9% for the Platanus, 14.8% for grasses, 9.8% for Olea and 3.6% for Plantago. The Platanus is one of the most frequently found pollens in the atmosphere over Madrid. At present, in this geographical area, a high percentage of patients with pollinosis are sensitive to this pollen. At least in some of these patients Platanus pollen is capable of inducing rhinitis and bronchial asthma.

Prediction of annual variations in atmospheric concentrations of grass pollen. A method based on meteorological factors and grain crop estimates.

We performed an aerobiologic observation of the grasses present in Madrid for 14 years (1978-1991), using volumetric air samplers. The counts obtained show that the major grass pollen release period (average daily grass pollen counts greater than 50 grains/m3 of air) occurs in the months of May and June, although lower counts can occur some days from the end of January onward. There are wide year-to-year variations in total atmospheric grass pollen counts, expressed as the total sum of the mean daily concentrations from April 1st to July 30th (ranging from 2568 to 6624). A strong, statistically significant correlation, based on Spearman's rank test and/or simple and multiple linear regressions, was found between the total grass seasonal count and preseasonal rainfall from October to March (R2 = 0.64; P = 0.0429). The meteorological variable which gave the correlation with greatest statistical significance (R2 = 0.97; P = 0.0016) was the average monthly preseasonal humidity from October to March. A good correlation was also found between March estimates of wheat, rye and barley crops and the total grass count (R2 = 0.73; P = 0.006). A model was designed from the above mentioned humidity variable through a multilinear regression analysis, and it was possible to predict, at the beginning of April, total seasonal counts for 1989 (predicted = 5468; actual = 4410; average error = 24%), 1990 (5033; 6090; -17%) and 1991 (3930; 2568; 53%). These data may help clinicians to predict and prepare themselves for the intensity of the grass pollen season and to explain yearly variations in the severity of symptoms.

Occupational asthma caused by Brazil ginseng dust.

The inhalation of different substances of plant origin can cause immediate and late onset asthma. The list of these agents responsible for such reactions is continuously increasing. We discuss a patient who developed symptoms of asthma after exposure to Pfaffia paniculata root powder used in the manufacturing of Brazil ginseng capsules. Airway hyperreactivity was confirmed by a positive bronchial challenge to methacholine. Sensitivity to this dust was confirmed by immediate skin test reactivity, a positive bronchial challenge (immediate response), and the presence of specific IgE detected by ELISA technique to an aqueous extract. The bronchial response was inhibited by sodium cromoglycate. Unexposed subjects did not exhibit reactivity to this ginseng extract with any of the tests referred to above. The same study performed with Korean ginseng (Panax ginseng) elicited negative results. This study is the first, to our knowledge, that links ginseng-root dust to occupational asthma.

Allergic conjunctivitis to chamomile tea.

Eye washing with chamomile tea is a folk remedy used by the general public to treat conjunctivitis and other ocular reactions. Chamomile is also found in many cosmetic products. Some cases of contact dermatitis (but not reactions of type I) were reported following its topical applications. We present seven hay fever patients that suffered from conjunctivitis; two of them also had lid angioedema after eye washing with chamomile tea. All seven patients had positive skin prick tests to the chamomile tea extract, Matricaria chamomilla pollen and Artemisia vulgaris pollen extracts. Positive conjunctival provocations were also observed in all the patients with the chamomile tea extract. In contrast, no symptoms were observed after oral challenges with this infusion. IgE activity against chamomile tea and Matricaria and Artemisia extracts (composite pollens) was detected by ELISA in the seven patients' sera. A cross reactivity among the above extracts was observed by an ELISA inhibition study. In all cases, the IgE activity to chamomile tea could fully be absorbed by Matricaria pollen extract. Skin prick tests and conjunctival provocation tests also performed in 100 hay fever controls revealed a positive immediate skin response to Artemisia in 15 patients, eight of them also to Matricaria pollen and five of them to Chamomile tea as well. Only two of the last patients had a positive conjunctival response. The results were negative in the rest of the controls. We conclude that the chamomile tea eye washing can induce allergic conjunctivitis. Matricaria chamomilla pollens contained in these infusions are the allergens responsible for these reactions.

Anaphylactic reaction after the ingestion of chamomile tea: a study of cross-reactivity with other composite pollens.

We report a case of an 8-year-old atopic boy in whom ingestion of a chamomile-tea infusion precipitated a severe anaphylactic reaction. The patient suffers from hay fever and bronchial asthma caused by a variety of pollens (grass, olive, and mugwort). This severe reaction was developed after his first ingestion of chamomile tea. Studies revealed the presence of immediate skin test reactivity and a positive passive transfer test to chamomile-tea extract. Moreover, both specific antichamomile-tea extract and anti-Matricaria chamomilla-pollen extract IgE antibodies were detected by an ELISA technique. Cross-reactivity among chamomile-tea extract and the pollens of Matricaria chamomilla, Ambrosia trifida (giant ragweed), and Artemisia vulgaris (mugwort), was demonstrated by an ELISA-inhibition study. These findings suggest a type I IgE-mediated immunologic mechanism as being responsible for the patient's anaphylactic symptoms and also suggest that the patient cross-reacted the pollens of Matricaria chamomilla contained in the chamomile tea because he was previously sensitized to Artemisia pollen.

Comparative functional study of colostral macrophages from mothers delivering preterm and at term.

Number and function of milk macrophages (MM) from 30 healthy mothers delivering preterm (mean gestational age 33 weeks) were compared with those obtained from 92 mothers with term delivery. The average concentration of MM in colostrum did not differ in the two groups: 0.55 x 10(6)/ml (preterm) and 0.42 x 10(6)/ml (term). Preterm MM showed the same activity as term MM in all three assayed functions (phagocytosis, chemotaxis and IL-1 secretion). Phagocytic activity, and impaired migratory motility and IL-1 secretion were significantly increased both in preterm and term MM in comparison with the activity in less mature blood monocytes. Thus it appears that preterm MM are, like term MM, a fully mature tissue macrophage population and therefore we suggest that with regard to MM both preterm and term colostrum are comparable, at least in the gestational age tested.