BACKGROUND:House dust mite (HDM) allergic rhinitis (HDM-AR) fluctuates over time with exposure conditions, making disease activity assessment essential when demonstrating treatment efficacy. We conducted post hoc tertile analyses to more accurately assess the efficacy of 300 IR HDM sublingual immunotherapy (SLIT) tablet in moderate-to-severe HDM-AR pediatric patients during periods of increased disease activity. METHODS:Data from two Japanese randomized controlled trials of 300 IR HDM SLIT-tablet were used to assess the average adjusted symptom score (AASS) and average combined symptom and medication score (ACSMS). To determine the impact of HDM-AR activity, placebo group scores during the primary period (end of treatment) at each center were ranked from lowest to highest to establish three tertiles. Scores differences between SLIT and placebo were analyzed in each tertile using ANCOVA. RESULTS:The first analysis included patients aged 5-16 years (300 IR = 193, placebo = 210). During the primary period, the effect of the SLIT-tablet, estimated at -13.1% (AASS) and -12.9% (ACSMS) versus placebo overall, was more pronounced in the highest tertile (relative differences -27.3% and -28.5%, respectively), with similar results across age groups. The second analysis including a pool of adolescents (300 IR = 120, placebo = 135) showed an overall effect of -19.9% (AASS) and -21.2% (ACSMS) with 300 IR versus placebo, with again greater improvements in the highest tertile (-32.5% and -34.1%, respectively). CONCLUSION:The greatest improvements occurred in the tertile where pediatric patients exhibited higher disease activity. This underscores the true efficacy of 300 IR HDM SLIT-tablet during periods when patients experience the most troublesome symptoms.

BACKGROUND:Highly effective CFTR modulators, particularly elexacaftor/tezacaftor/ivacaftor (ETI), produce rapid clinical improvements in people with cystic fibrosis. Yet early effects may be difficult to capture with spirometry or sweat test in patients with a mild disease or atypical mutations. Exhaled breath is rich in volatile organic compounds (VOCs) reflecting metabolic and inflammatory processes. We aimed to determine whether ETI induces early, measurable changes in breath composition and whether these relate to clinical outcomes. METHODS:Ten adults initiating ETI were enrolled in a prospective, open-label study with breath sampling at baseline, week one and month one. VOCs were measured using real-time proton-transfer-reaction - mass spectrometry (PTR-MS). Longitudinal changes were assessed using multilevel statistics, including univariate linear mixed-effects models, and multivariate repeated measures ANOVA-simultaneous component analysis plus (RM-ASCA+); repeated-measures correlations examined associations with lung function and sweat chloride concentration. Results were compared with a healthy cohort. RESULTS:Amongst the eight clinical responders, 11 features changed significantly after ETI initiation. Eight differed from healthy controls at baseline and shifted towards healthy levels over one month. RM-ASCA+ identified monotonous and non-monotonous patterns capturing various dynamics such as acute, progressive or delayed metabolic responses. A 11-feature PLS-DA model classified visits with high accuracy (AUC=0.84-0.96). Ten VOCs correlated with clinical readouts. Tentatively identified features pointed towards a shift in the microbiome and/or energy metabolism. CONCLUSIONS:ETI induces rapid alterations in exhaled VOCs, many trending towards healthy values and correlating with clinical improvement. Real-time breath analysis offers a promising non-invasive surrogate for early monitoring of therapeutic response.

Exhaled breath analysis represents a rapid and non-invasive strategy for detecting volatile organic compounds (VOCs) that mirror human metabolic activity and pathological processes. The soft ionization by chemical reaction in transfer (SICRIT) interface, when integrated with high-resolution mass spectrometry (HRMS), provides a flexible and powerful alternative to conventional analytical techniques for breath analysis. This study evaluated feasibility, analytical reproducibility, and chemical coverage of SICRIT-HRMS for clinical exhaled breath analysis. Signal processing included feature alignment, filtering and compound annotation through the Human Metabolome Database (HMDB) and the Human Breathomics Database (HBDB). In 40 healthy volunteers, SICRIT-HRMS reproducibly detected 604 spectral features (80% matching HMDB, 56% HBDB), including alkanes (oxidative stress markers) and semi-volatile amino acids (likely from airway microdroplets). Median reproducibility across exhalations was 13%, and cosine similarity between VOC profiles reached 0.98 within individuals. Pathway enrichment analysis revealed a significant contribution of amino-acid and lipid metabolism. Twenty-five VOCs were associated with gender and six with fasting duration. SICRIT-HRMS enables high-resolution, real-time profiling of the human breath metabolome with strong reproducibility and broad chemical coverage beyond classical VOCs. Its unique capabilities position it as a comprehensive tool for breathomics research and future translational diagnostic applications. ClinicalTrials.gov Identifier: NCT06020521.

BACKGROUND:Tablet formulations of allergen extracts are widely recommended over other formulations for the sublingual immunotherapy (SLIT) of respiratory allergies. However, with adequate clinical trial evidence, SLIT (liquid) drop formulations may be a relevant allergy treatment option. METHODS:The RHAPSODY multinational, Phase III, parallel-group, double-blind, placebo-controlled, randomised clinical study of adults with moderate-to-severe, grass-pollen-induced allergic rhinoconjunctivitis (ARC) with or without asthma was conducted at 45 investigating centres in six European countries. Participants received 26 months of continuous treatment with active 5-grass-pollen SLIT drops or placebo. The primary efficacy endpoint was the average daily total combined score (TCS, comprising a symptom score and a rescue medication score) during the second peak grass pollen season (PGPS). RESULTS:Of the 445 randomised patients (mean ± standard deviation (range) age: 32.6 ± 9.9 (18-63); males: 55.1%), 389 completed the trial. The primary efficacy endpoint showed a statistically significant difference in favour of active treatment versus placebo (average difference in the daily TCS: 1.88 (95% CI: 0.60-3.17); relative difference 26.51% (95% CI: 9.42-40.55); p = 0.0036). The difference (0.17 points) in the average weekly Rhinitis Quality of Life Questionnaire score during the second PGPS in favour of the active treatment was clinically relevant but not statistically significant. The differences in efficacy were generally driven by the medication score, rather than the symptom score. Most adverse events were mild and local. CONCLUSIONS:RHAPSODY was the first well-powered clinical trial to show the positive risk-benefit ratio of 5-grass-pollen SLIT drops in adult participants with moderate-to-severe grass-pollen-induced ARC.

Background Allergic rhinitis (AR) impacts quality of life, work and school productivity. Over the last years, an important body of evidence resulting from mHealth data has led to a better understanding of AR. Such advances have motivated an EAACI-endorsed update of the Allergic Rhinitis and its Impact on Asthma (ARIA) guidelines (ARIA 2024-2025). This manuscript presents the ARIA 2024-2025 recommendations for intranasal treatments, one of the mainstays for AR management.Methods The ARIA 2024-2025 guideline panel issued recommendations following the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) evidence-to-decision framework. Several sources of evidence were used to inform panel judgments and recommendations, including systematic reviews, evaluation of mHealth and pharmacovigilance data, as well as a survey of experts on costs.Results Eleven guideline questions concerning intranasal treatments for AR were prioritized, leading to recommendations. Overall, these questions concern the choice between different classes of intranasal medications-most notably, intranasal corticosteroids (INCS), antihistamines (INAH), fixed combinations of INAH+INCS and decongestants-or between different individual medications within each class. Four questions had not been evaluated in previous ARIA guidelines, while for the other three there was a change in the strength or directionality of recommendations. Overall, recommendations point to the suggested use of INAH+INCS over INAH or INCS and INCS over INAH.Conclusion This ARIA 2024-2025 article supports patients, their caregivers, and healthcare professionals in choosing an intranasal treatment. However, decisions on AR treatment should consider the clinical variability of the disease, patients' values, and the affordability of medications.

BACKGROUND:The triple combination Elexacaftor/Tezacaftor/Ivacaftor (ETI) translates into major respiratory improvements in adults; yet current clinical endpoints may prove insufficiently sensitive in young children. We hypothesised that ETI rapidly modifies the lungs' metabolism, resulting in changes in breath composition. METHODS:Eleven children with CF were enrolled in a longitudinal pilot study at the paediatric Necker hospital. Breath was collected on sorbent tubes using a ReCIVA® device before, after one week and one month of ETI. Samples were analysed by 2D-gas chromatography-mass spectrometry (2D-GC-MS). A linear mixed-effect model, corrected for clinical confounding factors, identified exhaled metabolites differentially expressed throughout the visits. Correlations were calculated between these and clinical indicators. RESULTS:Breath collection was successful in all children from six years old. They presented a decreased sweat chloride and improved lung function as early as within one week of ETI. Breath composition gradually evolved over the visits. ETI induced significant modifications in the level of 12 breath metabolites. Amongst those, dimethyl sulphide and tetradecene changes correlated with improvements in forced expiratory volume in one second (FEV1) and forced expiratory flow (FEF25-75), whilst 3-methyldecane and 3-(chloromethyl)-heptane were predictive of changes in lung clearance index (LCI2.5). CONCLUSIONS:ETI impacts the breath profile from the first week of treatment. Not only could "breathomics" bring mechanistic insights into the metabolic impact of ETI, but it may also offer novel non-invasive options to monitor CF disease and predict therapeutic response.

BACKGROUND:Patients with allergic rhinitis (AR) and/or mild or moderate asthma derived from birch-family pollen allergy can be treated with liquid sublingual immunotherapy (SLIT-liquid). This study evaluated the impact of two SLIT extracts on AR and asthma progression or onset in these patients. METHODS:This was a sub-analysis of a retrospective, longitudinal comparative cohort study that used a German prescription database. Patients treated with 3-tree (birch/alder/hazel) or birch-only SLIT-liquid and followed up for up to 6 years after treatment were compared with controls dispensed symptomatic medications. Multiple regression analysis compared dispensation data as a proxy for disease status and progression. RESULTS:A total of 493 patients treated with 3-tree SLIT-liquid and 311 treated with birch SLIT-liquid were analysed vs. 44,835 patients included as controls. Overall, 70.5 % of patients presented solely AR, 24.2 % solely asthma, and 5.3 % both diseases. Compared with controls, patients treated with 3-tree SLIT-liquid had reduced risk of AR [odds ratio (OR) = 3.21, 95 % CI 2.54-4.06, p < 0.001], asthma progression (OR = 2.03, 95 % CI 1.43-2.89, p < 0.0001), or asthma onset (OR = 0.592, 95 % CI, 0.408-0.860, p = 0.006). Birch-only SLIT-liquid showed similar effectiveness in reducing AR and asthma medication dispensation but no significant effect in reducing new-onset asthma. CONCLUSIONS:This real-world study demonstrated the effectiveness of treatment with 3-tree SLIT-liquid or birch SLIT-liquid in slowing the progression of birch-family pollen allergy. 3-tree SLIT-liquid covering a broader repertoire of epitopes mimicking natural exposure throughout the year may be valuable for patients sensitised to birch and/or alder and/or hazel pollen suffering from overlapping tree-pollen seasons.

Smoking is the main cause of chronic obstructive pulmonary disease (COPD) and is associated with corticosteroid resistance. Given the paucity of data on human lung preparations, macrophages (LMs), and parenchymal explants (LPEs) were exposed to cigarette smoke extracts (CSE) in the presence or absence of lipopolysaccharide (LPS). Moreover, LMs and LPEs were treated with budesonide prior exposure to CSE or LPS. The levels of cytokines (TNF-α, IL-6) and chemokines (CCL2, CCL4, CXCL1, CXCL5, and CXCL8) in the supernatants were measured using ELISAs. In LMs, exposure to CSE was not associated with significant difference in the production of cytokines and chemokines, with the notable exception of greater CXCL8 production. The results were generally the same for LPEs. CSE exposure did not potentiate the LPS-induced production of the cytokines and chemokines and even tended to reduce this production in LMs and LPEs. Lastly, CSE exposure inhibited budesonide's anti-inflammatory activity in LMs but not in LPEs. This study extends the data on the CSE inflammatory effects and its inhibition of corticosteroid efficacy in human lung preparations. Our findings question the relevance of these preparations with regard to the long-term toxicity of smoking and the corticosteroid resistance observed in smokers and in patients with COPD.

BACKGROUND:Adherence to rhinitis treatment has been insufficiently assessed. We aimed to use data from the MASK-air mHealth app to assess adherence to oral antihistamines (OAH), intra-nasal corticosteroids (INCS) or azelastine-fluticasone in patients with allergic rhinitis. METHODS:We included regular European MASK-air users with self-reported allergic rhinitis and reporting at least 1 day of OAH, INCS or azelastine-fluticasone. We assessed weeks during which patients answered the MASK-air questionnaire on all days. We restricted our analyses to data provided between January and June, to encompass the pollen seasons across the different assessed countries. We analysed symptoms using visual analogue scales (VASs) and the combined symptom-medication score (CSMS), performing stratified analyses by weekly adherence levels. Medication adherence was computed as the proportion of days in which patients reported rhinitis medication use. Sensitivity analyses were performed considering all weeks with at most 1 day of missing data and all months with at most 4 days of missing data. RESULTS:We assessed 8212 complete weeks (1361 users). Adherence (use of medication > 80% days) to specific drug classes ranged from 31.7% weeks for azelastine-fluticasone to 38.5% weeks for OAH. Similar adherence to rhinitis medication was found in users with or without self-reported asthma, except for INCS (better adherence in asthma patients). VAS and CSMS levels increased from no adherence to full adherence, except for INCS. A higher proportion of days with uncontrolled symptoms was observed in weeks with higher adherence. In full adherence weeks, 41.2% days reported rhinitis co-medication. The sensitivity analyses displayed similar results. CONCLUSIONS:A high adherence was found in patients reporting regular use of MASK-air. Different adherence patterns were found for INCS compared to OAH or azelastine-fluticasone that are likely to impact guidelines.

Background:Preclinical studies have recently revealed the critical role of innate immunity in determining lung transplantation outcomes. Although the International Society for Heart and Lung Transplantation recommends high-dose corticosteroid administration to donors, this practice is inconsistently applied worldwide. Investigating its impact on the donor lung's innate immune response - an unexplored area - could provide valuable evidence to support adoption of donor preconditioning with corticosteroids, beyond their traditional administration to recipients. Method:We used a cross-circulatory pig platform that consists of a donor lung placed extracorporeally and connected to the circulation of a recipient pig whose leukocytes are fluorescently labeled. Results:Donor preconditioning - compared to recipient's treatment alone - reduced the presence of CD3pos T-cells in the graft from both the donor and recipient, and enhanced the anti-inflammatory profile of alveolar macrophages, at least during the first 10 hours of donor-recipient interaction. The alveolar macrophages isolated from corticosteroid-preconditioned pig lungs exhibited decreased gene expression of T-cell-attracting chemokines during the 10-hour reperfusion period, correlating with the reduced T-cell infiltration. Similarly, human lung macrophages showed lower expression of these T-cell-attracting chemokines and higher anti-inflammatory profiles with corticosteroid treatment. Conclusion:Our results show that the early immune status of lung grafts is improved by treating donors with corticosteroids through macrophage-targeted mechanisms. This finding provides an immunological rationale for expanding the implementation of donor preconditioning with corticosteroids.

BACKGROUND:Pulmonary involvement (repeated lung infections, lung parenchymal inflammation, scarring, and malignancies) is frequent in patients with inborn errors of immunity (IEIs) and accounts for a significant proportion of the disease burden. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) can cure most severe IEIs. The indications for allo-HSCT have recently been extended to adults. OBJECTIVE:We sought to assess the impact of allo-HSCT specifically on respiratory status. METHODS:We retrospectively analyzed data of 50 patients with IEIs who underwent a first allo-HSCT after the age of 16 at 3 expert centers in France. RESULTS:The median length of follow-up was 4.8 years (interquartile range: 1.6-9.2) before allo-HSCT and 3 years (interquartile range: 1.4-6.0) after allo-HSCT. Ten patients died as a result of allo-HSCT-related complications. Four patients developed bronchiolitis obliterans syndrome. After 1-year posttransplantation, the mean annualized rate of severe respiratory infections (0.14 [95% CI: 0.04 to 0.24]) was lower than the value recorded before transplantation (0.54 [95% CI: 0.25 to 0.82]; P = .003 for paired comparisons of equivalent durations). Lung function was declining before allo-HSCT (mean FEV1: -2.09% predicted/year [95% CI: -7.27 to 3.09]) but increased afterward (+2.44% predicted/year [95% CI: -4.79 to 9.69], P = .0034 for paired comparisons). On computed tomography scans of the chest, bronchial disorders and lung parenchyma cavities were the most frequent abnormal findings. The bronchial thickening and bronchiolar micronodules regressed after allo-HSCT, whereas bronchiectasis and residual parenchymal cavities were stable. CONCLUSIONS:allo-HSCT seems likely to protect the long-term pulmonary prognosis of adults with IEIs; it is associated with a significantly lower incidence of severe respiratory infections, better lung function, and radiologic stabilization of lung damage.

BACKGROUND AND OBJECTIVE:The retrospective study EfficAPSI explored the real-world impact of liquid sublingual allergen immunotherapy (AIT; Staloral® SLIT-liquid) on health care resource utilization (HCRU) in allergic rhinitis (AR) patients with/without asthma. METHODS:In the EfficAPSI cohort, patients dispensed SLIT-liquid and AIT-naïve controls taking symptomatic drug treatment (SDT) were compared using propensity score weighting. A total of 5 periods were analyzed, namely, the historical pre-SLIT period (HP, 2 years before the index dose of SLIT/SDT [first dispensation]) and four 2-year follow-up periods (FUPs) after the index dose, with the latter 2 periods corresponding to post-treatment years. HCRU was analyzed using a Poisson model with generalized estimating equations. RESULTS:The study population comprised 112 492 SLIT and 333 082 control patients. Dispensations of antihistamines and intranasal corticosteroids decreased by 28% to 49% during the FUPs (IRR from 0.51 [0.50-0.52] to 0.69 [0.67-0.71]) and after treatment (IRR from 0.62 [0.59-0.65] to 0.72 [0.69-0.74]), favoring SLIT-exposed patients. In patients with asthma, a 17%-29% reduction in asthma medication dispensations also favored SLIT-liquid (IRR, 0.83 [0.78-0.88] to 0.71 [0.68-0.74] during treatment; 0.82 [0.77-0.88] to 0.78 [0.72-0.85] after treatment). For oral corticosteroids, the between-group difference in change from the HP was in favor of SLIT-liquid for all FUPs (IRR for doses, 0.66 [0.64-0.69] to 0.79 [0.73-0.85]). The decrease in medical consultations and hospitalizations was consistently more frequent over time in SLIT patients than in controls. CONCLUSIONS:In this national real-world study involving the largest number of person-years followed in the field of AIT to date, SLIT-liquid was associated with a reduction in AR and dispensation of asthma medication, including systemic corticosteroids, and medical consultations. The results recorded in the last 2 post-treatment FUPs suggest a sustained effect of SLIT-liquid.

AimThe role of somatostatin (SST) in the modulation of cholinergic neurotransmission has not been explored previously in human bronchi. We investigated the effects of SST, selective agonists of the five SST receptors SSTR, and octreotide (a SSTR2,3,5 agonist) on the cholinergic contraction induced in vitro either by acetylcholine or by electrical field stimulation (EFS) in human bronchial rings.MethodsHuman bronchial rings (n = 326) were obtained from 32 patients undergoing surgery for lung carcinoma. 5 Hz EFS (biphasic pulse width: 1 ms; constant current: 320 mA for 10 s) induced contractions that reached about ∼30% of the maximum contraction caused by 40 Hz EFS. Bronchial rings were stimulated for 240 min in the presence or absence of various concentrations of SST, octreotide, and selective agonists of each of the five SSTR receptors. Furthermore, the tissue and cellular locations of each of the five types of SSTR was determined by immunohistochemistry.ResultsSST, octreotide, and the SSTR agonists did not change the resting tone or the contractions produced by the cumulative addition of acetylcholine (10−9 to 10−3 M). In contrast, octreotide and the SSTR3 and SSTR5 agonists significantly increased the EFS-induced contractions. Immunoreactivity for all SSTR subtypes was detected in the airway’s neural ganglia.ConclusionThe present study provided new data on the location of SSTR in the human lung: notably, all types of receptor were found in the parasympathetic nerve ganglia of the bronchial wall. We suggest that the activation of prejunctional SSTR3 and SSTR5 receptors potentiates cholinergic-nerve-mediated contraction induced by EFS in human bronchi.

Background:Endothelial dysfunction (ED) is involved in the pathophysiology of idiopathic pulmonary fibrosis (IPF). Reactive hyperaemia peripheral arterial tonometry (RH-PAT) is a rapid, reproducible, noninvasive technique for assessing peripheral microvascular endothelial function. Methods:We conducted a prospective study of endothelial function (reactive hyperaemia index (RHI) as measured by RH-PAT) in patients with IPF, in order to explore the association between ED and respiratory symptoms, lung function and the disease prognosis. Adult patients with IPF underwent a baseline assessment of respiratory symptoms (including dyspnoea on the modified Medical Research Council scale), blood gas levels, pulmonary function tests, a 6-min walk test (6MWT) and endothelial function. The same examinations were performed 12, 24 and 36 months after inclusion. Results:42 patients with IPF (men: 76%) were enrolled. The median (IQR) age was 70 (61-75) years. The median (IQR) forced vital capacity and diffusing capacity of the lung for carbon monoxide (D LCO) were 77% (66-93%) and 43% (33-53%) predicted, respectively. The median (IQR) RHI was 1.86 (1.60-2.48). An ED defined at the RHI reference cut-off of 1.67 was observed in 13 (31%) patients. After 1 year of follow-up, there was a significant decline in endothelial function (median (IQR) difference: 0.65 (0.35-0.85), p=0.009). The RHI was associated with dyspnoea (r= -0.33, p=0.036), D LCO (r=0.36, p=0.025) and oxygen saturation at the end of 6MWT (r=0.33, p=0.049). The subgroups of patients with and without ED did not differ significantly in terms of mortality or the occurrence of acute exacerbations, although there was a nonsignificant trend to greater functional disease progression in patients with ED. Conclusion:Finger ED was correlated with respiratory symptoms and functional variables and worsened during the first year of follow-up. Our results suggest that the severity of IPF is associated with ED, although further studies are needed to assess the value of RH-PAT in the management of IPF.

The Allergic Rhinitis and its Impact on Asthma (ARIA) guidelines produced their first edition in 1999, with subsequent revisions in 2008, 2010, 2016 and 2019. A new iteration of ARIA-ARIA 2024-2025-in collaboration with EAACI is currently being developed, focusing on the management of allergic rhinitis. ARIA 2024-2025 follows the GRADE framework and is endorsed by the European Academy of Allergy and Clinical Immunology (EAACI). A set of approaches has been used to develop guideline questions, including surveying key opinion leaders and using artificial intelligence (AI)-based tools to analyse web searches on allergic rhinitis and to generate questions. Each prioritised guideline question is assessed through an Evidence-to-Decision (EtD) framework. EtDs support the systematic and transparent formulation of recommendations, comprising 12 criteria for which the best available evidence should be sought. In the context of ARIA-EAACI 2024-2025, such evidence is derived not only from randomised controlled trials but also-among others-from patient-generated data sources that better reflect the affected individuals' perspectives. Moreover, ARIA-EAACI 2024-2025 incorporates evidence on planetary health. Developed guideline recommendations will support the creation of digitalised decision algorithms and care pathways. This paper describes the methodology used to develop the person-centred, digitally enabled and AI-assisted ARIA-EAACI 2024-2025. Among others, it describes (i) the development and prioritisation of guideline questions, (ii) sources of evidence for EtDs and (iii) the development of digitalised decision algorithms and care pathways.

[This corrects the article DOI: 10.1016/j.lanepe.2024.100915.].

IntroductionLes pneumopathies interstitielles diffuses (PID) sont des pathologies rares d’étiologies variées menant pour la plupart à la fibrose pulmonaire. Sur une cohorte de 220 atteints de PID fibrosantes suivis à l’hôpital Foch, 58 présentaient un taux de polynucléaires à éosinophiles (PNE) sanguin>300/mm3 considéré comme élevé [1]. Bien que le rôle des PNE dans le développement et/ou l’exacerbation des PID ait été fortement suggéré [2], il n’a jamais été démontré. Nous émettons l’hypothèse que les PNE jouent un rôle dans la physiopathologie des PID. Afin de la vérifier, le projet INTREPID propose de caractériser les médiateurs de l’immunité de type 2 (cellulaires et solubles) chez 60 patients atteints de PID, en fonction de leur taux de PNE sanguin et de contrôler le caractère pro-fibrosant de leurs PNE in vitro. Les résultats présentés sont issus de la mise au point du modèle in vitro réalisés sur des volontaires sains.MéthodesLes PNE ont été isolés du sang de donneurs sains puis mis en culture, après vérification de leur pureté par cytométrie de flux CytoFlex SRT. Leur viabilité a été vérifiée par dosage de la lactate déshydrogénase (LDH) après 48h de culture. Les PNE ont été mis en coculture avec des lignées épithéliales bronchiques et alvéolaires (respectivement les cellules BEAS-2B et A549) en interface liquide-liquide et air-liquide. Les cytokines (IL-6, CXCL8, CCL5 et CCL2) ont été dosées par ELISA et une mesure de la résistance transépithéliale (TEER) a été effectuée. Les expectorations induites de sujets sains ont été analysées par cytométrie de flux et par coloration MGG afin d’étudier les différentes sous-populations cellulaires.RésultatsLes cultures de PNE présentent des caractéristiques satisfaisantes en terme de pureté (>90 %), absence d’activation des PNE (CD63-), ainsi qu’une viabilité jusqu’à 48h de culture (pas d’augmentation significative de la LDH). Lors des co-cultures en interface liquide-liquide, il a été observé une augmentation de la sécrétion de cytokines pro-inflammatoires (IL-6, CXCL8 et CCL2) par l’épithélium respiratoire BEAS-2B et A549 en présence de PNE. Une diminution de la TEER a également été observée en présence de PNE sur les cellules BEAS-2B. L’analyse par cytométrie des expectorations a permis d’identifier les différents types cellulaires tels que les macrophages, les monocytes, les éosinophiles, les neutrophiles, les lymphocytes T et les lymphocytes B.ConclusionCette étude de faisabilité décrit la mise au point de la culture des PNE et de la co-culture avec les lignées de cellules épithéliales respiratoires. Les modèles de coculture nous ont permis d’identifier un effet pro-inflammatoire des PNE au contact des lignées épithéliales bronchiques et alvéolaires. Par la suite ces expérimentations seront réalisées dans le cadre du projet INTREPID sur des PNE issus du sang et des expectorations induites de patients suivis pour une PID à l’hôpital Foch et des cellules primaires bronchiques humaines.