Variability of Total and Free IgE Levels and IgE Receptor Expression in Allergic Subjects in and Out of Pollen Season

The inter‐ and intra‐individual variability and seasonal variation of IgE, and high (FcεRI)‐ and low‐affinity (CD23) IgE receptor expression in blood of seasonal allergic rhinitis (SAR) subjects, is not well studied. Thirty‐two otherwise healthy subjects with a history of SAR to birch pollen and a positive skin prick test to birch pollen were sampled three times out of the pollen season and three times during the pollen season. FcεRI and CD23 expressions were analysed using flow cytometry. Total IgE was analysed using ImmunoCAP^® and free IgE was analysed with a novel customised research assay using an IgG‐FcεRI‐chimera protein coupled to ImmunoCAP as capture reagent, ImmunoCAP‐specific IgE conjugate and ImmunoCAP IgE calibrators. The performance of the free IgE assay was compared well with the reference ImmunoCAP total IgE assay. The working range of the assay was 0.35–200 kU/l IgE. FcεRI expression on basophils and CD23 expression on B cells showed low intrasubject variability both in and out of the pollen season (<10% CV). There was a small seasonal difference with lower total IgE levels (120 versus 128 kU/l; P = 0.004) and FcεRI expression (283 versus 325 mean fluorescence intensity (MFI); P < 0.001) during the pollen season. IgE, FcεRI expression and CD23 expression fulfilled biomarker and assay requirements of variability, and allergen exposure affected the biomarkers only to a minor degree. The free IgE assay may be used for measurement of free IgE levels in patients after anti‐IgE antibody treatment.     

Intranasal administration of allergen increases specific IgE whereas intranasal omalizumab does not increase serum IgE levels—A pilot study

Background

Administration of the therapeutic anti‐IgE antibody omalizumab to patients induces strong increases in IgE antibody levels.

Objective

To investigate the effect of intranasal administration of major birch pollen allergen Bet v 1, omalizumab or placebo on the levels of total and allergen‐specific IgE in patients with birch pollen allergy.

Methods

Based on the fact that intranasal allergen application induces rises of systemic allergen‐specific IgE, we performed a double‐blind placebo‐controlled pilot trial in which birch pollen allergic subjects were challenged intranasally with omalizumab, placebo or birch pollen allergen Bet v 1. Total and allergen‐specific IgE, IgG and basophil sensitivity were measured before and 8 weeks after challenge. For control purposes, total, allergen‐specific IgE levels and omalizumab‐IgE complexes as well as specific IgG levels were studied in subjects treated subcutaneously with either omalizumab or placebo. Effects of omalizumab on IgE production by IL‐4/anti‐CD40‐treated PBMCs from allergic patients were studied in vitro.

Results

Intranasal challenge with Bet v 1 induced increases in Bet v 1‐specific IgE levels by a median of 59.2%, and this change differed significantly from the other treatment groups (P = .016). No relevant change in allergen‐specific and total IgE levels was observed in subjects challenged with omalizumab. Addition of omalizumab did not enhance IL‐4/anti‐CD40‐induced IgE production in vitro. Significant rises in total IgE (mean IgE before: 131.83 kU/L to mean IgE after: 505.23 kU/L) and the presence of IgE‐omalizumab complexes were observed after subcutaneous administration of omalizumab.

Conclusion

Intranasal administration of allergen induced rises of allergen‐specific IgE levels, whereas intranasal administration of omalizumab did not enhance systemic total or allergen‐specific IgE levels.     

Immunotherapy in Hay Fever with Two Major Allergens 19, 25 and Partially Purified Extract of Timothy Grass Pollen

Grass pollen hay fever patients were hyposensitized in a prospective 3-year double blind study with two timothy extracts. Group WPA (20 patients) was treated with partially purified extract = whole pollen allergens (WPA) (Alutard®SQ) and group PPA (20 patients) was treated with a purified pollen allergen (PPA) = two isolated major allergens, Ag19 and Ag25. Both aluminiumhydroxide adsorbed extracts were biologically standardized. Clinical results from the first season have been recently published and WPA showed significantly fewer symptoms (P= 0.001) than PPA. Corresponding preseasonal and seasonal in vitro results are presented here. Serum total IgE, specific IgE versus total and individual allergens of timothy pollen and allergen-specific IgG showed a rapid increase in both groups until the season but showed no further increase or decrease during the season. Specific IgE was shown to correlate to specific IgG during hyposensitization. Group WPA, with fewer symptoms in grass pollen season than group PPA showed a significantly higher increase of specific IgG and IgE than group PPA, but individual symptom scores were not correlated to specific IgG or IgE, or their ratio. Specific IgE and IgG increases were not correlated to dosage. Surprisingly, almost all females were low-responders to specific IgG and IgE though they had equal symptom scores, dosage, and side effects as males, while the characteristics of high-responders to antibody were: youngest individuals and shortest duration of symptoms prior to treatment. Crossed radioimmunoelectrophoresis (CRIE) showed specific reaction to stimuli but no development of allergy against new timothy antigens. High response of IgE to Ag 19 in CRIE during initial hyposensitization seems suitable as marker for prospective evaluation of clinical effect in grass pollen hyposensitization. Nasal secretion was collected after methacholine provocation, and total IgE and specific IgE detected. There was no response to treatment, only a slight increase during the season. No decrease in nasal reactivity to methacholine was noted during one preseasonal hyposensitization.     

Evaluation of IgE antibodies to recombinant pollen allergens (Phl p 1, Phl p 2, and Phl p 5) in a random sample of patients with specific IgE to Phleum pratense

BACKGROUND: We measured specific IgE levels against the recombinant allergens (RAs) rPhl p 1, rPhl p 2, and rPhl p 5 in patients allergic to grass pollen, and examined the existence of different patterns of IgE production to RAs. The seasonal variations of IgE levels to rPhl p 1, rPhl p 2, and rPhl p 5 were considered, too. METHODS: Blood was taken from 276 consecutive patients with allergy to grass pollen diagnosed by patient history and skin prick testing. Total and specific serum IgE was measured by the immunoenzymatic CAP FEIA System. Eosinophil cationic protein (ECP) and myeloperoxidase (MPO) were assessed by radioimmunoassay according to the instructions of the manufacturers. RESULTS: We observed eight different patterns of IgE production to rPhl p 1, rPhl p 2, and rPhl p 5 in patients with specific IgE to timothy grass. A significant difference between the values of IgE levels to timothy and the sum of each level of specific IgE to individual RAs was found (P = 0.039, Wilcoxon matched pairs test) in the whole population (n = 276 subjects). In four subgroups of patients, the sum of each level of specific IgE to individual RAs was equal to the levels of specific IgE to timothy grass extract. In one subgroup, the sum of IgE to RAs was lower than the levels of IgE to the natural counterpart (P = 0.013). A lack of subjects in two subgroups did not permit comparison at all. Finally, three subjects with specific IgE to timothy did not show specific IgE to RAs. Out of 276 patients, blood was taken from two different groups of subjects at different time points: November-January and May-July, respectively. The median values were as follows: total IgE = 139 kU/l, IgE to timothy = 10.2 kUA/l; IgE to rPhl p 1 = 3.6 kUA/l, to rPhl p 2 = <0.35 kUA/l, and to rPhl p 5 = 1.1 kUA/l; ECP = 8.25 microg/l; MPO = 303.08 microg/l (before exposure to grass pollen); total IgE = 159 kU/l, IgE to timothy = 57.2 kUA/l; IgE to rPhl p 1 = 22.1 kUA/l, to rPhl p 2 = 5.9 kUA/l, and to rPhl p 5 = 3.9 kUA/l; ECP = 16.21 microg/l; MPO = 413.09 microg/l (during the pollen season). There were significant variations of specific IgE levels between the patients exposed to pollen and the unexposed patients. Moreover, there were statistical differences in the IgE, ECP, and MPO levels in sera before and during the pollen season P<0.035, P<0.017, and P<0.0062, respectively. CONCLUSIONS: The results suggest that RAs allow establishment of the patient's IgE-reactivity profile, encourage future research, and encourage manufacturers to produce further RAs for precise diagnosis and substantially improved immunotherapy injection of only those allergens against which significant amounts of specific IgE are produced.     

IgE antibodies to Hymenoptera venoms in the serum are common in the general population and are related to indications of atopy

Determination of Hymenoptera venom (HV)-specific serum IgE antibodies is a useful diagnostic method in patients with systemic anaphylactic reaction (SAR) to Hymenoptera stings. In a general population cohort, we determined the prevalence of SAR and HV-specific IgE antibodies and assessed parameters associated with the latter. A total of 277 voluntarily participating inhabitants of rural Bavaria (Germany) (232 adults, mean age 38.0 years; 45 children, mean age 8.4 years) were investigated for a history of atopic disease or SAR to insect stings; in 258 of these, total IgE and specific IgE antibodies to HV ( Apis mellifera, Vespula vulgaris/germanica ) and four common aeroallergens (birch pollen, grass pollen, house-dust mite, and cat dander) in the serum were determined. Nine (3.3%) subjects reported SAR to insect stings. In 27.1% of the sera, specific IgE antibodies to HV were found, to bee venom in 24.8%, and to wasp venom in 8.5% ( P <0.0001). Of those exhibiting HV-specific IgE, 7.1% reported SAR to insect stings. A personal history of atopic disease (hay fever, asthma, or atopic eczema) was present in 16.7%, specific IgE to common aeroallergens was found in 32.6%, and total IgE> 100 kU/1 was found in 22.5%. Specific serum IgE to HV was significantly associated with male sex (female vs. male, OR = 0.47; CI 0.25–0.86), young age (children vs. adults, OR = 2.80; CI 1.25–6.28), a history of SAR to insect stings (OR = 4.16; CI 1.15–15.03), total slgE>100kU/l (OR = 3.88; CI 1.98–7.60), and specific IgE antibodies to three of the four aeroallergens (grass pollen, OR = 7.24 CI 3.66–14.38; birch pollen, OR = 3.67 CI 1.54–8.81; and house-dust mite, OR = 4.61 CI 2.08–10.32). It is concluded that immunologic sensitization to HV is common in the general population and is associated with atopy-related humoral IgE hyperresponsiveness.     

Efficacy and safety of 5-grass pollen sublingual immunotherapy tablets in patients with different clinical profiles of allergic rhinoconjunctivitis

Background The optimal dose of grass pollen tablets for sublingual immunotherapy (SLIT) in allergic rhinoconjunctivitis patients was previously established in a multinational, randomized, double‐blind, placebo‐controlled study in 628 adults. Patients were randomized to receive once‐daily 5‐grass pollen sublingual tablets of 100 IR (index of reactivity), 300 IR or 500 IR, or placebo starting 4 months before the pollen season. Objective The aim of this complementary analysis was to determine whether 300 IR 5‐grass pollen SLIT‐tablets is effective in different subtypes of patients who are allergic to grass pollen. Methods Different subgroups could be identified regarding comorbidities (with or without asthma during the grass‐pollen season), sensitization (mono/polysensitization) and symptom severity. An additional exploratory analysis was performed within four subgroups based on pre‐treatment assessment: Group 1=high specific IgE; Group 2=high symptom scores; Group 3=high skin sensitivity; Group 4=any of Group 1, 2 or 3. Results Asthma and sensitization status were not significant covariates as the average Rhinoconjunctivitis Total Symptom Score (RTSS) was identical for patients with and without grass‐pollen asthma, as well as for mono‐ and polysensitized patients. Across the four subgroups, average RTSSs (± SD) for the optimal dosage (300 IR) were 3.91 ± 3.16, 3.83 ± 3.14, 2.55 ± 2.13 and 3.61 ± 2.97, for subgroups 1, 2, 3 and 4, respectively. ancova showed that in Group 1 average RTSS did not differ significantly with different doses of SLIT. In Groups 2, 3 and 4, doses of 300 IR and 500 IR were significantly more effective than 100 IR and placebo (P0.035). All doses of SLIT administered in this study can be considered safe in the patients investigated. Conclusions The risk–benefit ratio validates the use of 300 IR tablets in clinical practice in all of these patient subgroups, regardless of severity profile, sensitization status and presence of asthma.     

Seasonal idiopathic rhinitis with local inflammatory response and specific IgE in absence of systemic response

Background: Patients with idiopathic rhinitis (IR) are considered to be nonallergic because they have a negative skin prick test (SPT) and allergen specific‐IgE in serum. The concept of localized mucosal allergy in the absence of atopy has recently been proposed. The immunological mechanisms involved in seasonal IR have not been sufficiently studied. We examined nasal mucosa inflammation, the presence of nasal specific‐IgE and the response to nasal allergen provocation test (NAPT) in patients with seasonal IR who presented symptoms only in spring. Methods: We evaluated 32 patients with seasonal IR and 35 with persistent allergic rhinitis to pollen (PAR‐P) and compared these with healthy controls and persons with PAR to house dust mite during the pollen season, as well as by NAPT out‐of‐season with grass and Olea europea. We measured the nasal leukocyte–lymphocyte phenotype (CD45, CD33, CD16, CD3, CD4 and CD8), eosinophil‐cationic‐protein, and total and specific‐IgE to grass and olive pollen in serum and nasal lavage and performed NAPT. Results: In the IR group, 62.5% had a positive NAPT (IR‐PosNAPT), 20/32 to grass, with four of these having a positive NAPT to olive pollen as well. IR‐PosNAPT patients showed a similar nasal leukocyte–lymphocyte profile to the PAR‐P patients and different to controls. We detected nasal specific‐IgE in 35% of IR‐PosNAPT patients. Conclusions: These results support the hypothesis that a subgroup of patients with IR have seasonal symptoms with evidence of a nasal allergic immune reaction in the absence of a positive SPT or serum specific IgE.     

Omalizumab is more effective than suplatast tosilate in the treatment of Japanese cedar pollen-induced seasonal allergic rhinitis

Background Seasonal allergic rhinitis (SAR) induced by Japanese cedar pollens is a major problem in Japan. Omalizumab, a humanized monoclonal anti‐IgE antibody, improves symptoms associated with SAR, but a comparative study with an anti‐allergy drug has not yet been conducted. Objective To compare the efficacy and safety of omalizumab with suplatast tosilate, a selective T‐helper type 2 (Th2) cytokine inhibitor, in patients with Japanese cedar pollen‐induced SAR. Methods A randomized, double‐blind, double‐dummy study was conducted in 308 Japanese patients with a history of moderate‐to‐severe SAR who showed a CAP‐RAST value (2+) specifically to Japanese cedar pollens. Patients were treated for 12 weeks with omalizumab plus placebo of suplatast tosilate or suplatast tosilate plus placebo of omalizumab. Results The mean daily nasal symptom medication scores (sum of the daily nasal symptom severity score and daily nasal rescue medication score) were significantly lower in the omalizumab group than in the suplatast tosilate group during three evaluation periods (P<0.001). The omalizumab group also had significantly lower mean daily nasal severity scores, each of the mean daily nasal and ocular symptom severity scores (sneezing, runny nose, stuffy nose, itchy nose, itchy eyes, watery eyes, and red eyes). Omalizumab reduced rescue medication requirements, and the proportion of days with any rescue medication use in the omalizumab group was significantly lower. Serum‐free IgE levels markedly decreased in the omalizumab group and it was associated with clinical efficacy. The adverse reaction profiles were similar between the two groups. The overall incidence of injection site reactions was higher in the omalizumab group than in the suplatast tosilate group, but all these events were of mild degree. No anti‐omalizumab antibodies were detected. Conclusion Omalizumab showed significantly greater improvements than suplatast tosilate in the treatment of SAR induced by Japanese cedar pollens.     

Detection of allergen-specific IgE in tears of grass pollen-allergic patients with allergic rhinoconjunctivitis

Background Allergic conjunctivitis is a common symptom amongst Type I (IgE-mediated) allergic diseases; and mosl frequently seen as rhinoconjunctivitis. However, the site of production and the significance of allergen specific IgE needs further elucidation. Objective We investigated whether the presence of IgE in tears of grass pollen allergic patients correlated with disease and clinical symptoms, whether the IgE binding pattern to the different grass pollen antigens was diflferent in sera and tears, and whether IgA antibodies to grass pollen allergens were present in tears. Finally, we looked whether specific IgE was produced locally or was exudated from serum. Methods Sera from 44 grass pollen allergic patients suffering from either allergic rhinitis (n=11) or allergic rhinoconjunctivitis (n= 33) and from healthy controls (n= l) were used for the experiments. Binding of specific IgE and IgA antibodies to the differyent groups of grass pollen allergens (Phleum pratense) was evaluated by means of immunobtotting. Results Specific IgE was detected in sera as well as in tears of allergic patients, whereby tear-derived allergen-specific IgE exerted similar specificities to the corresponding IgE from serum. The correlation between symptoms of ocular allergy and the presence of allergen-specific IgE in tears was highly significant (P 0.0001). In contrast, only a poor correlation was found between specific and/or total IgE in sera and the manifestation of ocular allergy (P = 0.73). Conclusion Allergen-specific IgE antibodies in tears seem to be produced locally rather than exsudated from serum. IgE in tears seems to be responsible for allergic conjunctivitis. IgA in tears cannot exert a protective function since the IgA antibodies recognize different antigens in a grass pollen (Phleum pratense) extract than IgE antibodies. The highly significant correlation between allergic conjunctivitis and the presence of specific tear IgE emphasizes the diagnostic value of immunoblots with tear IgE, especially in cases in which serum provides inconclusive results.     

Basophil histamine release, IgE, eosinophil counts, ECP, and EPX are related to the severity of symptoms in seasonal allergic rhinitis

BACKGROUND: Serum specific IgE, basophil histamine release, and blood eosinophil parameters are associated with allergic rhinitis, but investigations of the relationship to the severity of allergic symptoms are few and conflicting. Our study aimed to investigate the seasonal changes in the following laboratory tests: specific IgE, basophil histamine release, eosinophil counts, and serum and plasma eosinophil cationic protein (ECP) and eosinophil protein X (EPX), and to analyze, in detail, the relationship of each individual test to the severity of symptoms in rhinitis patients allergic to both birch and grass pollen. METHODS: The above tests were performed on blood samples obtained from 49 allergic rhinitis patients during the birch-pollen season, during the grass-pollen season, and after the seasons. Symptom-medication diaries were filled in during both pollen seasons. We used partial least square (PLS) analysis to establish an optimal statistical link between the symptom score and medication and the laboratory tests, in an investigator-independent way. RESULTS: Increases in specific IgE, basophil histamine release, eosinophil counts, serum ECP and EPX, and plasma EPX were observed from the birch-pollen season to the grass-pollen season, followed by a decrease from the grass-pollen season to after the pollen seasons, except for the specific IgE. No seasonal changes in plasma ECP and total IgE were seen. The PLS analysis found a relationship between symptom score and medication and the aggregate laboratory tests (F-test value 40.2, correlation 0.34 for the cumulative relation). However, the variation in laboratory tests could explain only half of the total variation in symptoms and less than a quarter of the total variation in medication. The symptom score and, to a minor degree, medication were especially correlated with the basophil histamine-release results, with a decreasing relevance of specific IgE, eosinophil counts, total IgE, serum and plasma EPX, and serum ECP. Plasma ECP was not related to the symptom score and medication. CONCLUSIONS: A significant relationship between the severity of allergic rhinitis and various allergic inflammatory markers was found but could account for only a minor part of the variation in the patients' evaluation of their disease.     

The allergen profile of beech and oak pollen

Background Beech and oak pollen are potential allergen sources with a world‐wide distribution. Objective We aimed to characterize the allergen profile of beech and oak pollen and to study cross‐reactivities with birch and grass pollen allergens. Methods Sera from tree pollen‐allergic patients with evidence for beech and oak pollen sensitization from Basel, Switzerland, (n=23) and sera from birch pollen‐allergic patients from Vienna, Austria, (n=26) were compared in immunoblot experiments for IgE reactivity to birch (Betula pendula syn. verrucosa), beech (Fagus sylvatica) and oak (Quercus alba) pollen allergens. Subsequently, beech and oak pollen allergens were characterized by IgE inhibition experiments with purified recombinant and natural allergens and with allergen‐specific antibody probes. Birch‐, beech‐ and oak pollen‐specific IgE levels were determined by ELISA. Results Beech and oak pollen contain allergens that cross‐react with the birch pollen allergens Bet v 1, Bet v 2 and Bet v 4 and with the berberine bridge enzyme‐like allergen Phl p 4 from timothy grass pollen. Sera from Swiss and Austrian patients exhibited similar IgE reactivity profiles to birch, beech and oak pollen extracts. IgE levels to beech and oak pollen allergens were lower than those to birch pollen allergens. Conclusion IgE reactivity to beech pollen is mainly due to cross‐reactivity with birch pollen allergens, and a Phl p 4‐like molecule represented another predominant IgE‐reactive structure in oak pollen. The characterization of beech and oak pollen allergens and their cross‐reactivity is important for the diagnosis and treatment of beech and oak pollen allergy.     

Can component‐based microarray replace fluorescent enzimoimmunoassay in the diagnosis of grass and cypress pollen allergy?

Background Few data on the diagnostic accuracy in pollinosis of the microarray ISAC of allergens are available. Objective We aim to comparatively analyse ISAC CRD103 with the whole‐extract ImmunoCAP in grass and cypress pollen allergy, evaluating the suitability of the manufacturer's recommended cut‐off points for both techniques. Methods We studied 120 atopic patients grouped into grass and cypress pollen‐allergic patients and controls based on clinical history and skin prick tests. Specific IgE against Phleum pratense and Cupressus arizonica by ImmunoCAP and ISAC CRD103 were performed on all subjects. Results In the grass pollen group (43 allergic/26 controls), both microarray and CAP showed high sensitivity (Se) and specificity (Sp) values (ISAC: Se 97.7, Sp 92.3; CAP: Se 95.3, Sp 96.1) for recommended cut‐off points. Comparing the optimal (ISAC: 0.4 ISU; CAP: 0.33 kU/L) with the recommended cut‐off points within the same technique, diagnostic agreement was observed in both techniques. Thus, CAP and ISAC showed similar diagnostic performance in grass pollen allergy when using recommended cut‐off points. In cypress pollen group (12 allergic/92 controls), the microarray (Se: 91.7, Sp 91.3) showed similar Se but significantly higher Sp (P=0.034) than CAP (Se: 91.7, Sp: 80.4) using recommended cut‐off points. However, although diagnostic performance of the microarray did not change when comparing the optimal (0.82 ISU) with the recommended cut‐off point, CAP improved diagnosis of cypress pollen allergy, when applying the optimal (0.66 kU/L)(CAP Se: 91.7, Sp: 89.1) instead of the manufacturer's recommended cut‐off point. Thus, when the most suitable cut‐off point for both techniques (ISAC: 0.3 ISU; CAP: 0.66 kU/L) is selected, microarray and CAP provide equivalent diagnoses. Conclusions and Clinical Relevance Component‐based microarray ISAC CRD103 and whole‐allergen CAP showed high Se and Sp diagnosing equally grass and cypress pollen allergy. The cut‐off point for each allergen should be properly applied for both techniques. Cite this as: P. Cabrera‐Freitag, M. J. Goikoetxea, C. Beorlegui, P. Gamboa, G. Gastaminza, M. Fernández‐Benítez, M. Ferrer, M. Blanca and M. L. Sanz, Clinical & Experimental Allergy, 2011 (41) 1440–1446.     

Clinical significance of total IgE and of specific IgE to Dermatophagoides spp., grass pollen and other common allergens II. Relationship to clinical manifestations

The relationships of total and specific IgE levels to the clinical manifestations and atopic status of ninety-two subjects, suffering from asthma, rhinitis and infantile eczema in varying combinations, were examined. No significant correlations were found between specific IgE levels to Dermatophagoides spp. and grass pollen or between total IgE levels, and the age of the patient, nor the age of onset, duration, severity and frequency of symptoms. A highly significant correlation was found between total IgE levels and the numbers of skin test reactions to a battery of twenty routine tests and to the sum total of the graded sizes of the reactions in each subject.     

Oral delivery of Lactobacillus casei Shirota modifies allergen-induced immune responses in allergic rhinitis

Background Changes in the composition of the gut microbiota have been implicated in the pathogenesis of allergic disorders, suggesting beneficial interactions between the intestinal immune system and specific bacterial strains. Lactobacilli are naturally present within the complex gastrointestinal microbiota of humans and they are currently present in many probiotic supplements.
Objective We sought to investigate the role that Lactobacillus casei Shirota (LcS) may play in modulating seasonal allergic rhinitis (SAR).
Methods The study format was double-blinded, placebo-controlled with 10 SAR sufferers in each group. We have documented and compared changes in immune status arising through the daily ingestion of a milk drink with or without live LcS, over a period of 5 months. Pre-, peak- and post-grass pollen season blood samples were collected for determination of plasma total IgE and grass pollen-specific IgG and IgE levels by an enzyme immunoassay. At the same time, cytokine levels were determined by flow cytometric bead array technology following culture of peripheral blood mononuclear cells for 6 days in the presence or absence of specific grass pollen antigens.
Results Volunteers treated with LcS showed a significant reduction in levels of antigen-induced IL-5, IL-6 and IFN-γ production compared with volunteers supplemented with placebo. Meanwhile, levels of specific IgG increased and IgE decreased in the probiotic group.
Conclusion Changes in antigen-induced production of cytokines were observed in patients treated with probiotics. These data show that probiotic supplementation modulates immune responses in allergic rhinitis and may have the potential to alleviate the severity of symptoms.     

Induction of Bronchial Tolerance After 1 Cycle of Monophosphoryl-A-Adjuvanted Specific Immunotherapy in Children With Grass Pollen Allergies

PurposeSubcutaneous allergen-specific immunotherapy (SCIT) is a well-established and clinically effective method to treat allergic diseases, such as rhinitis and asthma. It remains unclear how soon after initiation of an ultra-short course of grass pollen immunotherapy adjuvanted with monophosphoryl lipid A (MPL)-specific bronchial tolerance can be induced.MethodsIn a prospective study of 69 children double-sensitized to birch and grass pollens (51 males, average age 11.1 years), development of bronchial tolerance after 1 cycle of SCIT for grass was evaluated. In all the patients, the bronchial allergen provocation test (BAP) was performed before and after treatment. According to the results of the first BAP, the patients were divided into 2 groups: those showing a negative BAP with a decrease in FEV1 of <20% (seasonal allergic rhinitis [SAR] group, n=47); and those showing a positive BAP with a decrease in FEV1 of ≥20% (SAR with allergic asthma [SAR and Asthma] group, n=22). All the patients received MPL-adjuvanted, ultra-short course immunotherapy for birch, but only those with a positive BAP to grass received MPL-SCIT for grass.ResultsAfter the pollen season, the BAP in the SAR group remained unchanged, while it was improved in the SAR and Asthma group (decrease in FEV1 of 28.8% vs 12.5%, P<0.01). The IgG4 levels increased after SCIT (median before SCIT 0.34 to 11.4 after SCIT), whereas the total and specific IgE levels remained unchanged.ConclusionsAfter 1 cycle of MPL-SCIT, specific bronchial tolerance may be significantly induced, whereas in patients without SCIT, bronchial hyperactivity may remain unchanged.     

Effects of IL-4 and IL-13 on total and allergen specific IgE production by cultured PBMC from allergic patients determined with recombinant pollen allergens

Background: Interleukin (IL)-4 and IL-13 have been shown to be potent switch factors for IgE synthesis in human B cells. Objective: In this study we investigated the effects of recombinant human IL-4 and IL-13 on total and allergen specific IgE synthesis by peripheral blood inononuclear colls (PBMC) from pollen allergic patients and healthy control individuals. Methods: Peripheral blood mononuclear cells (PBMC) from allergic patients were investigated for their capacity to produce allergen specific IgE in vitro. Total protein extracts from birch pollen and timothy grass pollen as well as purified recombinant birch pollen allergens, Bet v I, birch profiling (Bet v II) and recombinant timothy grass pollen allergens. Phi p I, Phi p II, and Phi p V were used to measure specific IgE-antibody synthesis in cell culture supernatants by IgE-immunoblot and ELISA. Reults PBMC obtained from allergic patients spontaneously secreted allergen specific IgE in the culture supernatants. Addition of Interleukin 4, Interleukin 13 and anti-CD40 antibody to the cultures alone or in combinations significantly induced total IgE production whereas allergen specific IgE production was not affected. Conclusion: Our results indicate that the peripheral blood of allergic individuals contains long lived allergen specific B cells which have already switched to IgE production and which are not sensitive to IL-4 and IL-13 treatment. These results may have implications on attempts to use cytokines or cytokine antagonists in therapy of Type I allergy.     

Immunological features of asthma (Part II)

A serological comparison was made of two groups of 120 matched asthmatic and healthy subjects, between the ages of 20 and 49 years and matched for age and sex, in terms of serum total levels of IgG, IgM, IgA. IgD and IgE and of specific antibody levels in each immunoglobulin class to five common UK allergens. The relationship of clinical features to the serological tests was also examined in the asthmatic subjects. The following statistically significant findings were shown. The patients had higher levels than the controls of total globulins and of IgG, IgA and IgD but not IgM. In both patients and controls the females had higher IgM levels than the males. The total IgE levels were higher in patients than in the controls and the male patients had higher levels than the female patients. Total IgE levels were also related, to the numbers of first degree relatives with asthma, hay fever and eczema, to the severity of hay fever and to the amount of time off work in the male patients. In those male patients with exercise induced asthma the total IgE levels were lower than in those not showing this reaction. As for the other iminunoglobulins, the only significant differences were a higher [gG level in patients with FEV, or PFR>50% predicted and a higher IgD level in patients with hay fever. Radio-immunodiffusion tests for specific precipitins were positive for Dermatophagoldes pteronyssinus in comparable numbers of asthmatics (25.8%) and controls (21.7%). Positive precipitin tests were uncommon in tests with extracts of grass pollen, Aspergillus fumigatus, cat and dog hair in the patients and even less so in the controls. Positive RAST tests for specific IgE antibodies were obtained in patients and controls respectively, against D. pteronyssinus 59% and 11% grass pollen 37.0 and 12%, and A. fumigatus 6% and 4%. The male patients showed the closest significant relationship of specific IgE to D pteronyssinus and the history of house dust allergy, positive skin test and nasal test. in the females only the skin and specific IgE tests were related. Both sexes showed a significant association between specific IgE to grass pollen and positive skin tests and nasal tests, but only the males showed an association with the history. The size of skin test weal to D. pteronyssinus were related to the levels of specific IgE antibody, Correspondence: Professor J. Pepys, Cardiothoracic Institute, Brompton, London SW3 6HP. No differences were found between the four skin test groups and between the asthmatics and the control subjects in the incidence of bacterial precipitins and auin-antibodies.     

Childhood asthma: clinical and immunological changes over a decade

A group of 26 Australian asthmatic children with laboratory-proven bronchial hyper-reactivity to the allergens of rye grass pollen and/or the house dust mite has been studied over a 9-year period. Clinical symptoms and drug scores were used to evaluate the severity of the patients' asthma and, wherever possible, blood samples were obtained before, during and after the rye grass pollen seasons. The cumulative symptom and drug scores for the 20 patients with bronchial hyper-reactivity to rye grass pollen extract tended to increase during and fall after each pollen season but the peaks were of decreasing amplitude over the 9 years. Since a proportion of these patients underwent hyposensitization to rye grass during year 1, longitudinal comparisons were made between year 2 and year 9. Comparing the individuals at the same three seasonal time-points revealed significantly lower drug scores in year 9 compared with year 2, and in parallel with this, significantly lower total IgE, IgE anti-rye and IgG anti-rye antibodies at all three assessment points. In the 14 patients with bronchial hyper-reactivity to house dust mite the severity of the asthma and the median levels of IgE and IgG mite specific antibodies all decreased over the study period. Despite the progressive improvement in asthma and diminishing immune responses to both rye grass and house mite in the patients, no immunological feature could be identified that correlated significantly with clinical outcome.     

A Case of Occupational Rhinitis Induced by Maize Pollen Exposure in a Farmer: Detection of IgE-Binding Components

Corn is a major staple food, along with rice and wheat, in many parts of the world. There are several reports of hypersensitivity to maize pollen. However, cases of occupational allergic rhinitis induced by inhalation of maize pollen are very rare. We herein report the case of a 67-year-old male with occupational rhinitis caused by occupational exposure to maize pollen in a cornfield. He showed positive responses to maize pollen, as well as grass pollens, in skin prick tests. A high level of serum immunoglobulin E (IgE) specific to maize pollen extracts was detected by an enzyme-linked immunosorbent assay (ELISA). Laboratory tests showed a high serum level of total IgE (724 kU/L) and a high level of IgE specific to maize pollen (8.32 kU/L) using the Immuno-CAP system. Occupational rhinitis was confirmed by a nasal provocation test with maize pollen extracts. IgE ELISA inhibition tests showed antibody cross-reactivity between maize pollen and grass pollen extracts. IgE immunoblotting using maize pollen extracts demonstrated a 27 kDa IgE-binding component. These findings suggest that maize pollen can induce IgE-mediated occupational rhinitis in exposed workers.     

Basophil histamine release in patients with hay fever. Results compared with specific IgE and total IgE during immunotherapy.

Histamine release from leucocytes was demonstrated in grass pollen hay fever patients on in vitro challenge with extract of Pleum pratense (timothy). No release was found in persons without a history of grass pollen allergy. During preseasonal hyposensitization the following tendencies were found in cell sensitivity to allergen as well as in specific IgE antibody level of serum: an initial increase at the beginning of the therapy followed by a decrease during the pollen season. This is in contrast to untreated hay fever patients in whom an increase or no change at all of cell sensitivity and specific IgE was observed in the pollen season. Immunotherapy, therefore, can prevent such an increase in the pollen season. The mechanism might be due to a depression of the IgE production. In untreated as well as in treated patients the cell sensitivity was found to be significantly correlated to the grass specific IgE determined by RAST but not to the total serum level of IgE estimated by RIST. It seems likely that the sensitivity would be useful for evaluating the degree of allergy in grass pollen hay fever patients treated or not treated with immunotherapy.