Chronic oral corticosteroids use and persistent eosinophilia in severe asthmatics from the Belgian severe asthma registry.

BACKGROUND: Severe asthma (SA) may require frequent courses or chronic use of oral corticosteroids (OCS), inducing many known side effects and complications. Therefore, it is important to identify risk factors of chronic use of OCS in SA, considering the heterogeneity of clinical and inflammatory asthma phenotypes. Another aim of the present analysis is to characterize a subpopulation of severe asthmatics, in whom blood eosinophil counts (BEC) remain elevated despite chronic OCS treatment. METHODS: In a cross-sectional analysis of 982 SA patients enrolled in the Belgian Severe Asthma Registry (BSAR) between March 2009 and February 2019, we investigated the characteristics of the OCS treated patients with special attention to their inflammatory profile. RESULTS: At enrollment, 211 (21%) SA patients were taking maintenance OCS (median dose: 8 [IQR: 5-10]) mg prednisone equivalent). BEC was high (> 400/mm3) in 44% of the OCS treated population. Multivariable logistic regression analysis showed that risk factors for chronic use of OCS in SA were late-onset asthma (i.e. age of onset > 40 yr), frequent exacerbations (i.e. ≥2 exacerbations in the previous year) and non-atopic asthma. Late-onset asthma was also a predictor for persistently high BEC in OCS treated SA patients. CONCLUSION: These data showed a significant association between a persistently high BEC and late-onset asthma in OCS treated SA patients. Whether it is poor compliance to treatment or corticosteroid insensitivity the reasons for this association warrants further investigation.

Heterogeneity of phenotypes in severe asthmatics. The Belgian Severe Asthma Registry (BSAR).

UNLABELLED: The Belgian severe asthma registry is a web-based registry encompassing demographic, clinical, functional and inflammatory data of severe asthmatics (SA), aiming at improving awareness, knowledge on its natural history and subphenotypes, and offering tools to optimize care of this asthma population. METHODS: The cross-sectional analyses of this registry included 350 SA as defined by the ATS (2000) from 9 Belgian centres, with at least one year follow up. RESULTS: Mean age was 55 ± 14 yrs. SA were more frequently female (57%) and atopic (70%). Late-onset asthma (≥40 yr) was observed in 31% of SA. Current smokers represented 12% while 31% were ex-smokers. In addition to high doses ICS + LABA, 65% of patients were receiving LTRA, 27% anti-IgE and 24% maintenance oral corticosteroids (8 mg (Interquartile range-IQR:4-8) methylprednisolone). Despite impaired airflow (median FEV1:67%; IQR: 52-81) only 65% had a post-bronchodilator FEV1/FVC ratio <70%. The median blood eosinophil count was 240/mm³. The median FENO was 26 ppb (IQR: 15-43) and 22% of SA had FENO ≥ 50 ppb. Induced sputum was successful in 86 patients. Eosinophilic asthma (sputum Eos ≥ 3%) was the predominant phenotype (55%) while neutrophilic (sputum Neu ≥ 76%) and paucigranulocytic asthma accounted for 22% and 17% respectively. Comorbidities included rhinitis and chronic rhinosinusitis (49%), nasal polyposis (19%), oesophageal reflux (36%), overweight and obesity (47%) and depression (19%). In addition, 8% had aspirin-induced asthma and 3% ABPA. Asthma was not well-controlled in 83% according to ACT < 20 and 77% with ACQ > 1.5. CONCLUSION: In this cohort of patients with severe asthma, the majority displayed indices of persistent airflow limitation and eosinophilic inflammation despite high-dose corticosteroids, suggesting potential for eosinophil-targeted biotherapies.

The impact of concomitant rhinitis on asthma-related quality of life and asthma control.

BACKGROUND: Characterizing the interactions between the upper and lower airways is important for the management of asthma. This study aimed at assessing the specific impact of concomitant rhinitis on asthma-related quality of life (QOL) and asthma control. METHODS: A cross-sectional, observational survey was conducted among 1173 patients with asthma (aged 12-45) recruited by general practitioners and chest physicians. AR was defined by self-reported rhinitis symptoms and previously documented sensitization to inhalant allergens. The primary outcomes were (1) asthma control assessed by the Asthma Control Questionnaire (ACQ) and (2) asthma-specific QOL evaluated through the Mini Asthma Quality of Life Questionnaire (mAQLQ). RESULTS: AR was present in 73.9% of the population with asthma and nonallergic rhinitis (NAR) in 13.6%. AR and NAR were associated with an increased risk of uncontrolled asthma (i.e. ACQ score > 1.5) with adjusted odds ratios (OR) of 2.00 (95% confidence interval [CI]: 1.35-2.97) and 1.77 (95%CI: 1.09-2.89), respectively. Multivariate linear regression analysis showed that AR and NAR had a modest, although significant, negative impact on the global mAQLQ score (beta coefficient: -0.293, standard error [SE]: 0.063 and beta coefficient: -0.221, SE: 0.080, P < 0.001, respectively), even after adjustment for the level of asthma control and demographic characteristics. CONCLUSION: This survey provides direct evidence that AR and NAR are associated with an incremental adverse impact on the disease-specific QOL of patients with asthma and the level of asthma control. Further investigations are required to determine whether appropriate treatment of rhinitis would efficiently reduce asthma morbidity.

A comparison of budesonide/formoterol maintenance and reliever therapy vs. conventional best practice in asthma management.

OBJECTIVE: To study the effectiveness and safety of budesonide/formoterol (Symbicort) Maintenance And Reliever Therapy (Symbicort SMART, AstraZeneca, Södertalje, Sweden), a simplified management approach with one inhaler compared with conventional best practice (CBP) with multiple inhalers in patients with persistent asthma. DESIGN: Open-label randomised controlled parallel group trial, 6-month treatment. PARTICIPANTS: A total of 908 patients > or = 12 years of age, with persistent asthma receiving treatment with inhaled corticosteroids (ICS), either alone or in conjunction with long-acting beta(2)-agonist. MAIN OUTCOME MEASURES: Time to first severe asthma exacerbation and number of severe asthma exacerbations. RESULTS: No difference between groups was seen in time to first severe exacerbation (p = 0.75). Exacerbation rates were low in both groups. A total of 12 patients in the Symbicort SMART group experienced a total of 14 severe asthma exacerbations, and 19 patients in the CBP group experienced a total of 25 severe asthma exacerbations (annual rate 0.07 vs. 0.13 p = 0.09). The mean daily dose of ICS expressed in BDP equivalent was significantly lower in the Symbicort SMART group (including as-needed use) vs. in the CBP group (749 microg vs. 1059 microg; p < 0.0001). Mean scores in Asthma Control Questionnaire, 5 question version improved significantly in the SMART group compared with the CBP group (p = 0.0026). Symbicort SMART and CBP were equally well tolerated. The mean drug cost/patient/month was significantly lower for the patients in the Symbicort SMART group compared with patients receiving CBP (51.3 euros vs. 66.5 euros; p < 0.0001). CONCLUSIONS: In Belgian patients, a simplified regimen using budesonide/formoterol maintenance and reliever therapy was at least as effective at improving clinical control compared with CBP with a significantly lower ICS dose and significantly lower drug costs.

"Real-life" effectiveness of omalizumab in patients with severe persistent allergic asthma: The PERSIST study.

OBJECTIVE: To evaluate the 16- and 52-week effectiveness of add-on omalizumab treatment under real-life heterogeneity in patients, settings, and physicians in an open-label, multicenter, pharmaco-epidemiologic study of patients with severe persistent allergic asthma in Belgium. METHODS: Effectiveness outcomes included improvement in 2005 global initiative for asthma (GINA) classification, physician-rated global evaluation of treatment effectiveness (GETE), quality of life (Juniper asthma-related quality of life (AQLQ) and European quality of life questionnaire 5 dimensions (EQ-5D)), and severe asthma exacerbations. Patients studied included both intent-to-treat and per-protocol populations. RESULTS: The sample (n=158) had a mean age of 48.17+/-17.18 years, and a slight majority were female (53.8%). Despite being treated with high-dose inhaled corticosteroids and long-acting beta2-agonists, all patients experienced frequent symptoms and had exacerbations in the past year. At 16 weeks, >82% had good/excellent GETE (P values <0.001), >82% had an improvement in total AQLQ scores of > or =0.5 points (P<0.001), and >91% were severe exacerbation-free (P<0.001). At 52 weeks, >72% had a good/excellent GETE rating (P<0.001), >84% had improvements in total AQLQ score of > or =0.5 points (P<0.001), >56% had minimally important improvements in EQ-5D utility scores (P=0.012), and >65% were severe exacerbation-free (P<0.001). Significant reductions in healthcare utilization compared to the one year prior to treatment were noted. CONCLUSION: The PERSIST study shows better physician-rated effectiveness, greater improvements in quality of life, greater reductions in exacerbation rates, and greater reductions in healthcare utilization than previously reported in efficacy studies. Under real-life conditions, omalizumab is effective as add-on therapy in the treatment of patients with persistent severe allergic asthma.

Exhaled nitric oxide as a marker of asthma control in smoking patients.

Exhaled nitric oxide fraction (F(eNO)), which is a reliable marker of eosinophilic airway inflammation, is partially suppressed by tobacco smoking. Consequently, its potential as a biomarker in asthma management has never been evaluated in smoking patients. In the present study, the authors tested the validity of F(eNO) to predict asthma control in this population. F(eNO) and the Asthma Control Questionnaire (ACQ) were recorded at least once in 411 nonsmoking (345 with at least two visits) and 59 smoking (51 with at least two visits) asthma patients. Despite similar mean ACQ scores (1.5 versus 1.7), F(eNO) was reduced in smoking asthmatics (18.1 ppb versus 33.7 ppb). A decrease in F(eNO) of <20% precludes asthma control improvement in nonsmoking (negative predictive value (NPV) 78%) and in smoking patients (NPV 72%). An increase in F(eNO) <30% is unlikely to be associated with deterioration in asthma control in both groups of patients (NPV = 86% and 84% in nonsmoking and smoking patients, respectively). It is concluded that, even in smokers, sequential changes in F(eNO) have a relationship with asthma control. The present study is the first to indicate that cigarette smoking does not obviate the clinical value of measuring F(eNO) in asthma among smokers.

Exhaled nitric oxide and asthma control: a longitudinal study in unselected patients.

Controlled studies have shown that monitoring of the exhaled nitric oxide fraction (F(eNO)) improves asthma management. However, the studies seldom consider the full range of patients seen in clinical practise. In the present study, the ability of F(eNO) to reflect asthma control over time is investigated in a regular clinical setting, and meaningful F(eNO) cut-off points and changes are identified. Answers to the Asthma Control Questionnaire and F(eNO) were recorded at least once in 341 unselected asthma patients. The whole population and subgroups were considered, i.e. both inhaled corticosteroid (ICS)-naïve and low or high-to-medium (500 mug beclomethasone dipropionate equivalents.day(-1)) ICS-dose groups. An F(eNO) decrease <40% or increase <30% precludes asthma control optimisation or deterioration, respectively (negative predictive value 79 and 82%, respectively). In the present study's low-dose group, a decrease >40% indicated asthma control optimisation (positive predictive value (PPV) 83%). In ICS-naïve patients, F(eNO) >35 ppb predicted asthma control improvement in response to ICS (PPV 68%). In most cases, forced expiratory volume in one second assessments were not useful. In conclusion, in a given patient, exhaled nitric oxide fraction was found to be significantly related to asthma control over time. The overall ability of exhaled nitric oxide fraction to reflect asthma control was reduced in patients using high doses of inhaled corticosteroids. Forced expiratory volume in one second had little additional value in assessing asthma control.

[Omalizumab, an anti-IgE monoclonal antibody to treat severe asthma]. / L'omalizumab (Xolair), un anticorps monoclonal anti-IgE pour traiter l'asthme sévère.

Omalizumab (Xolair) is the first representative of a new therapeutical class for severe allergic asthma. By neutralizing Ac IgE, omalizumab fulfils an anti-inflammatory action of which the effect has been shown beneficial in the treatment of severe allergic asthma and particularly in severe asthma for which the therapeutical arsenal is for the time being disappointing and associated to frequent side effects there where omalizumab is well tolerated.

[Omalizumab, an anti-IgE monoclonal antibody to treat severe asthma]. / L'omalizumab (Xolair), un anticorps monoclonal anti-IgE pour traiter l'asthme sévère.

Omalizumab (Xolair) is the first representative of a new therapeutical class, which will be soon available in severe allergic asthma. By neutralizing Ac IgE, omalizumab fulfils an anti-inflammatory action of which the effect has been shown beneficial in the treatment of severe allergic asthma and particularly in severe asthma for which the therapeutical arsenal is for the time being disappointing and associated to frequent side effects there where omalizumab is well tolerated.

[The pneumology department]. / Le Service de Pneumologie.

The department of pneumology of the Erasme hospital exists since 25 years. The basic clinical activities include pulmonary function testing (7,500 patients per year), endoscopy, including interventional endoscopy (1,500 patients per year), thoracic oncology, allergology, rehabilitation and aid to smoking cessation. The following expertise fields have been largely developed: lung transplantation, treatment of cystic fibrosis in collaboration with the children's hospital Reine Fabiola, occupational.

Increased expression of high but not low molecular weight heat shock proteins in resectable lung carcinoma.

Strong expression of high-molecular-weight (HMW) heat-shock proteins (HSP) by lung carcinoma has been documented using immunohistochemistry. Far less is known about the expression of low-molecular-weight (LMW) HSP in lung cancer. We compared the quantitative expression of HMW (HSP-60, HSP-70) and LMW (HSP-27, ubiquitin) HSP in tumor and non-tumor lung tissue obtained from 47 patients undergoing surgical resection of lung carcinoma. HSP levels were determined in cell lysates from tissue samples by ELISA using streptavidin-biotin technology. Results were normalized to total protein content measured by spectrophotometry. Compared to disease-free lung tissue, tumor tissue samples showed higher levels of both HSP-60 (median value: 227 pg versus 96 pg per mg protein (P<0.001 by Wilcoxon Rank test for paired data) and HSP-70 (median value: 525 ng versus 401 ng per mg protein (P=0.01 by Wilcoxon Rank test for paired data). Tumor and tumor-free tissues show similar levels of ubiquitin and HSP-27. Neither the survival rate nor the histologic type and extent of cancer are correlated with the observed differences in HSP-60 and HSP-70 expression (P>0.1 by one way analysis of variance for repeated measures with one between subject factor). Our data confirm, on a quantitative basis, the increased expression of HSP-60 and HSP-70 in non-small-cell lung carcinoma. However, no prognostic value was found to be associated with this over-expression. In contrast, LMW stress proteins such as ubiquitin and HSP-27, although implicated in cellular processes potentially related to malignant transformation, show no increased expression in lung carcinoma.

Early effect of ultrarush venom immunotherapy on the IgG antibody response.

BACKGROUND: We have previously shown in several allergy models that allergic and tolerance status with respect to allergens is associated with a somewhat different dominant specificity of IgG antibodies. The objective was to test this hypothesis in the compelling model of ultrarush venom immunotherapy (VIT), which induces clinical tolerance after only a few hours of treatment. METHODS: Antibody titers and specificity were evaluated through solid-phase ELISA using streptavidin-biotin technology in 12 patients allergic to wasp venom before and during the ultrarush procedure (at 12 h, 24 h, and 15 days). The results were compared with those from another group of 20 patients treated with venom injections for at least 2 years. RESULTS: No significant change was observed in IgG titers during the early phase of VIT. The capacity of individual sera to prevent the antigen binding of pooled IgG from allergic patients changed rapidly, with mean percentage inhibitions falling from 80+/-15%, before starting VIT, to 26+/-14%, 35+/-15%, and 34+/-5% after 12 h, 24 h, and 15 days of treatment, respectively (P<0.001 by one-way ANOVA). The capacity of individual sera to prevent the antigen binding of pooled IgG from patients receiving prolonged VIT changed, with mean percent inhibitions increasing from 47+/-8%, before starting VIT, to 76+/-7%, 83+/-6%, and 87+/-6% after 12 h, 24 h, and 15 days of treatment, respectively (P<0.001 by one-way ANOVA). CONCLUSIONS: During the initial phase of ultrarush VIT, a change in IgG specificity, i.e., a change in the set of epitopes dominantly recognized by IgG on wasp-venom antigens, occurred concomitantly with early clinical tolerance and was already detectable a few hours after the onset of treatment. Although it may be an epiphenomenon, this change represents the earliest humoral modification described so far during this procedure. The mechanism is unknown, but it appears to be a selective depletion of the highest avidity antibody fraction by the venom injected in large doses at this stage of therapy. Finally, our data now show the previously documented association between a particular IgG specificity and the clinical status (allergy vs tolerance) to be true also with ultrarush VIT, a model in which the clinical ability to display allergic symptoms is rapidly reversed.

Helper T-cell responses elicited by Der p 1-pulsed dendritic cells and recombinant IL-12 in atopic and healthy subjects.

BACKGROUND: Environmental allergens, such as Dermatophagoides pteronyssinus group 1 antigen (Der p 1), induce T(H2)-type responses in atopic patients, whereas healthy individuals have T(H1)-type responses to the same antigens. Because of their efficient synthesis of IL-12, dendritic cells (DCs) are potent inducers of T(H1)-type immune responses. OBJECTIVE: We sought to determine whether DCs would skew allergen-specific T(H2)-type responses from atopic individuals. METHODS: Purified CD4(+) T cells from healthy donors or atopic individuals were cultured in the absence or presence of recombinant (r)IL-12 with DCs derived from PBMCs and pulsed with Der p 1. Supernatants of DC-T cell cocultures were assayed by ELISA for IL-5 and IFN-gamma. RESULTS: A T(H1)-type response developed in purified CD4(+) T cells from healthy donors in response to Der p 1-pulsed DCs, as indicated by high levels of IFN-gamma in culture supernatants. In contrast, CD4(+) T cells from atopic donors displayed a T(H2)-type profile characterized by high levels of IL-5 and low levels of IFN-gamma. The addition of rIL-12 (10 ng/mL) to DC-T cell cocultures resulted in the induction of IFN-gamma secretion by Der p 1-specific CD4(+) T cells from atopic patients, whereas their production of IL-5 was not inhibited. Using flow cytometry after intracytoplasmic staining, we found that IFN-gamma and IL-5 were secreted by distinct CD4(+) T-cell subpopulations. CONCLUSION: The cytokine profile of Der p 1-specific T(H2)-like cells from atopic individuals is maintained when the allergen is presented by DCs, even in the presence of exogenous rIL-12.

Relationship between allergic status and specificity of IgG antibody to inhaled allergens: the grass pollen model.

BACKGROUND: We have previously reported that IgG antibodies from healthy individuals and patients suffering from non-seasonal mite allergy bind to different sets of epitopes on Der p 1, allowing almost complete discrimination of the populations. OBJECTIVES: To confirm this observation in a seasonal allergy model where a clear relationship between allergic symptoms and exposure to the offending agent is established. To investigate whether the pattern of modified specificity is related to the differences in IgG subclass hierarchy usually exhibited by nonallergic and allergic populations. METHODS: The capacity of individual sera from patients allergic to grass pollen and healthy individuals, including grass pollen-sensitized subjects, to prevent the binding of pooled IgG, IgG1, and IgG4 fractions from grass pollen-allergic patients and healthy individuals to solid-phase bound grass pollen antigen was evaluated in enzyme-linked immunosorbent assay (ELISA) using streptavidin-biotin technology. Specificity controls were performed using sera from patients allergic to cat dander and house dust mite. RESULTS: The capacity of sera to prevent the antigen binding of allergic IgG averaged 84 +/- 5% for allergic sera and 53 +/- 6% for healthy sera (P < 0.001 by one-way anova). Conversely, using the antigen-binding capacity of healthy control IgG as reference, percentage inhibitions averaged 46 +/- 9% in grass pollen-allergic subjects compared with 80 +/- 4%, 82 +/- 2% in healthy individuals, and mite- and cat-allergic patients, respectively, resulting in two well-separated populations (P < 0.0001 by one-way anova). Similar results were found regardless of whether pooled IgG1 or IgG4 were used. CONCLUSION: Together with previous data, our results define a new type of humoral signature in the immune response to inhaled allergens. Allergic and healthy status differ not only in the presence or absence of specific IgE antibody but also in the preferential expression of distinct IgG specificities that are better correlated with clinical manifestations and are unrelated to subclass distribution.

Anti-betalactoglobulin IgG antibodies bind to a specific profile of epitopes when patients are allergic to cow's milk proteins.

BACKGROUND: We demonstrated recently that mite-allergic patients differed from healthy controls in the specificity of their IgG antibodies towards mite antigens. OBJECTIVE: The present study investigates whether these discriminatory IgG responses could be associated with the expression and the evolution of clinical manifestations in allergy to cow's milk proteins. METHODS: Antibody specificity was evaluated by comparing IgG-binding to native bovine beta-lactoglobulin (nBLG) and its products of pepsin hydrolysis (dBLG) using a solid-phase enzyme-linked immunosorbent assay (ELISA). Antibody specificity was further investigated in competitive ELISA using streptavidin-biotin technology with purified IgG fractions from selected subjects and specific mouse monoclonals raised against BLG. RESULTS: IgG antibodies from CM-intolerant or allergic sera (n=222) showed a higher degree of binding to nBLG than to dBLG, while control sera showed similar levels to both nBLG and dBLG (n=99 children/65 adults). Sera from symptomatic patients, wether or not they contained IgE antibodies, demonstrated group-segregating capacities to compete with pooled purified IgG from each clinical class, and with selected murine anti-nBLG monoclonal antibodies for binding to n- and dBLG. Furthermore, this inhibitory capacity shifted dramatically in a small subset (n=14) of children as they developed CM-tolerance. CONCLUSIONS: The IgG responses to BLG of CM-intolerant or allergic patients are very different from those of healthy controls, being characterized not only by increased titres but also similar patterns of modified specificity, including a marked preference for conformational epitopes. Cross-competition experiments confirmed that the restricted specificity was clinically associated, appearing as an immunological signature, which allowed almost complete discrimination between patient groups. This phenomenon is a particularly promising diagnostic feature in this category of young patients where conventional tests usually only document the status of sensitization.

Modified antigenic reactivity of anti-phospholipase A2 IgG antibodies in patients allergic to bee venom: conversion with immunotherapy and relation to subclass expression.

BACKGROUND: We have previously reported that, in addition to modifying IgG levels and subclass distributions, wasp venom immunotherapy (VIT) rapidly changes IgG antibody specificity. OBJECTIVES: We investigated whether such a change can be documented in the IgG response to the major bee venom allergen, phospholipase A2 (PLA2), from patients allergic to bees treated with VIT; whether it is coupled to the shift in IgG subclass distribution (IgG4 predominance) usually observed during VIT; and whether it restores the specificity displayed by IgG antibodies from nonallergic individuals. METHODS: Antibody specificity was evaluated in 17 patients allergic to bee venom in competitive ELISAs by using streptavidin biotin technology. Patients were tested before and during specific immunotherapy (at 15 days and 6 months) and compared with another group of 17 patients treated with venom injections for at least 2 years (VIT patients) and 30 healthy individuals. RESULTS: The capacity of individual sera to prevent PLA2 binding of pooled IgG from allergic patients changed rapidly with mean percentage inhibitions falling from 84% +/- 14% before starting VIT to 27% +/- 13% and 28% +/- 7% after 15 days and 6 months of treatment, respectively (p < 0.001 by one-way analysis of variance [ANOVA]). IgG titers were only slightly increased. The capacity of individual sera to prevent the binding of pooled IgG from patients receiving VIT changed rapidly with mean percentage inhibition increasing from 60% +/- 12% before starting VIT to 85% +/- 6% and 82% +/- 6% after 15 days and 6 months of treatment, respectively (p < 0.001 by one-way ANOVA). Similar results were found regardless of whether pooled IgG1 or pooled IgG4 were used. CONCLUSION: VIT results in a rapid change in the antigenic reactivity of anti-PLA2 IgG antibody of human allergic sera, restoring, although not completely, the specificity peculiar to lgG from healthy individuals. This suggests that allergic status and immunoprotection correlate with the preferential expression of distinct IgG specificities, which appear equally distributed over the IgG1 and IgG4 antibody subclasses. It is, however, not known whether the shift in IgG specificity is one of the operative mechanisms of VIT.

Mite allergy is associated with a specific profile of IgG epitopes recognized on antigen p1 of Dermatophagoides pteronyssinus.

BACKGROUND: Immune response to inhaled antigens differs in allergic patients and healthy individuals, mostly in the quality of T cell help provided (i.e. Th2 or Th1 dominant subset). However, while different in many functional aspects, both groups of T cells shared the capacity to support the synthesis of antigen-specific immunoglobulin G (IgG) antibodies detected in different amounts in the serum of atopic and healthy individuals. OBJECTIVE: The present study investigates whether these IgG responses display similar or different epitopic dominance in the mite sensitization model. METHODS: Antibody specificity was evaluated by comparing the IgG binding to native Der p1 (nDer p1) and its products of pepsin hydrolysis (dDer p1) in 56 mite-allergic patients and 148 healthy individuals, including 24 mite-sensitized individuals in a solid enzyme linked immunosorbent assay (ELISA). Antibody specificity was also studied in competitive ELISA using streptavidin biotin technology. RESULTS: Mite-allergic sera showed a higher degree of binding to nDer p1 than to dDer p1, whereas control sera and mite-sensitized sera bound at a similar level to the two forms of the antigen. Allergic sera and control sera, including mite-sensitized sera, showed distinct capacities to prevent the binding to nDer p1 of pooled IgG from each group as well as murine monoclonal antibodies specific to Der p1. CONCLUSION: The IgG response to Der p1 of mite-allergic patients differs from that of healthy controls and mite-sensitized subjects, not only in its increased titres but also in its consistant pattern of modified specificity, displaying a marked preference for conformational epitopes. Cross-competition experiments confirm the clinically associated, restricted specificity, allowing almost complete discrimination between groups, particularly between mite-sensitized and mite-allergic subjects, which is currently impossible with routinely available assays.

Wasp venom immunotherapy changes IgG antibody specificity.

BACKGROUND: The evolution of the IgG response during venom immunotherapy (VIT) has been previously investigated in terms of antibody titres and subclasses. OBJECTIVES: The present work studied the evolution of IgG antibody fine specificity in wasp allergic patients treated with rush VIT. METHODS: Antibody specificity was evaluated in 51 wasp allergic patients in competitive ELISA using streptavidin biotin technology. Patients were tested before and during specific rush immunotherapy (at 15 days, 6 months, 12 months) and compared with 44 patients treated by venom injections for at least 2 years. RESULTS: The capacity of sera to prevent the antigen binding of pooled IgG from allergic patients changed rapidly with mean percentage inhibitions (+/-SD) falling from 70+/-11-51+/-18% after 15 days of treatment (P<0.001 by one way ANOVA). Similarly, the antigen binding capacity of pooled IgG from VIT patients was differently prevented by sera with mean percentage inhibitions increasing from 37+/-12-65+/-8 after 15 days of treatment (P< 0.0001 by one-way ANOVA). CONCLUSIONS: The immunodominance pattern of IgG epitopes recognized on wasp venom antigens by sera from wasp allergic patients changes soon after initiating rush VIT. Further studies will indicate whether, instead of measuring IgG titres, this marked change could be used as the basis of a new test for monitoring the outcome of VIT.