T Cells Promote Bronchial Epithelial Cell Secretion of Matrix Metalloproteinase-9 via a C-C Chemokine Receptor Type 2 Pathway: Implications for Chronic Lung Allograft Dysfunction.

Chronic lung allograft dysfunction (CLAD) is the major limitation of long-term survival after lung transplantation. CLAD manifests as bronchiolitis obliterans syndrome (BOS) or restrictive allograft syndrome (RAS). Alloimmune reactions and epithelial-to-mesenchymal transition have been suggested in BOS. However, little is known regarding the role of allogenicity in epithelial cell differentiation. Primary human bronchial epithelial cells (BECs) were treated with activated T cells in the presence or absence of transforming growth factor (TGF)-ß. The expression of epithelial and mesenchymal markers was investigated. The secretion of inflammatory cytokines and matrix metalloproteinase (MMP)-9 was measured in culture supernatants and in plasma from lung transplant recipients (LTRs): 49 stable, 29 with BOS, and 16 with RAS. We demonstrated that C-C motif chemokine 2 secreted by T cells supports TGF-ß-induced MMP-9 production by BECs after binding to C-C chemokine receptor type 2. Longitudinal investigation in LTRs revealed a rise in plasma MMP-9 before CLAD onset. Multivariate analysis showed that plasma MMP-9 was independently associated with BOS (odds ratio [OR] = 6.19, p = 0.002) or RAS (OR = 3.9, p = 0.024) and predicted the occurrence of CLAD 12 months before the functional diagnosis. Thus, immune cells support airway remodeling through the production of MMP-9. Plasma MMP-9 is a potential predictive biomarker of CLAD.

TLR3 promotes MMP-9 production in primary human airway epithelial cells through Wnt/ß-catenin signaling.

BACKGROUND: Airway epithelial cells (AEC) act as the first line of defence in case of lung infections. They constitute a physical barrier against pathogens and they participate in the initiation of the immune response. Yet, the modalities of pathogen recognition by AEC and the consequences on the epithelial barrier remain poorly documented. METHOD: We investigated the response of primary human AEC to viral (polyinosinic-polycytidylic acid, poly(I:C)) and bacterial (lipopolysaccharide, LPS) stimulations in combination with the lung remodeling factor Transforming Growth Factor-ß (TGF-ß). RESULTS: We showed a strong production of pro-inflammatory cytokines (Interleukin (IL)-6, Tumor Necrosis Factor α, TNFα) or chemokines (CCL2, CCL3, CCL4, CXCL10, CXCL11) by AEC stimulated with poly(I:C). Cytokine and chemokine production, except CXCL10, was Toll Like Receptor (TLR)-3 dependent and although they express TLR4, we found no cytokine production after LPS stimulation. Poly(I:C), but not LPS, synergised with TGF-ß for the production of matrix metalloproteinase-9 (MMP-9) and fibronectin. Mechanistic analyses suggest the secretion of Wnt ligands by AEC along with a degradation of the cellular junctions after poly(I:C) exposure, leading to the release of ß-catenin from the cell membrane and stimulation of the Wnt/ß-catenin pathway. CONCLUSION: Our results highlight the cross talk between TGF-ß and TLR signaling in bronchial epithelium and its impact on the remodeling process.

[Transbronchial lung cryobiopsy in interstitial lung diseases]. / Les cryobiopsies pulmonaires transbronchiques dans les pneumopathies infiltrantes diffuses.

In some cases of interstitial lung disease (ILD), clinical and biological findings associated with CT scan pattern during multidisciplinary discussion (MDD) fail to yield a confident diagnosis. In these cases, histology may be necessary. Transbronchial lung cryobiopsy (TBLC) is a bronchoscopic procedure that has been developed in recent years and currently contributes to diagnostic work-up in patients with ILD. TBLC provides tissue samples for histological analysis with an acceptable risk of complications, consisting mainly in pneumothorax or bleeding. In addition to higher diagnostic yield than conventional forceps biopsies, the procedure shows a better safety profile than surgical biopsies. The indication to perform TBLC is decided during a 1st MDD and during a 2nd MDD, results can provide a diagnostic yield approximating 80%. TBLC appears to be an attractive, minimally invasive technique to be proposed as a first-line procedure in selected patients in experienced centers, while surgical lung biopsy may be considered as a second-line solution.

Controlled donation after circulatory death lung transplantation: Results of the French protocol including in situ abdominal normothermic regional perfusion and ex vivo lung perfusion.

BACKGROUND: The French national protocol for controlled donation after circulatory determination of death (cDCD) includes normothermic regional perfusion (NRP) in case of abdominal organ procurement and additional ex-vivo lung perfusion (EVLP) before considering lung transplantation (LT). METHODS: We made a retrospective study of a prospective registry that included all donors considered for cDCD LT from the beginning of the program in May 2016 to November 2021. RESULTS: One hundred grafts from 14 donor hospitals were accepted by 6 LT centers. The median duration of the agonal phase was 20 minutes [2-166]. The median duration from circulatory arrest to pulmonary flush was 62 minutes [20-90]. Ten lung grafts were not retrieved due to prolonged agonal phases (n = 3), failure of NRP insertion (n = 5), or poor in situ evaluation (n = 2). The remaining 90 lung grafts were all evaluated on EVLP, with a conversion rate of 84% and a cDCD transplantation rate of 76%. The median total preservation time was 707 minutes [543-1038]. Seventy-one bilateral LTs and 5 single LTs were performed for chronic obstructive pulmonary disease (n = 29), pulmonary fibrosis (n = 21), cystic fibrosis (n = 15), pulmonary hypertension (n = 8), graft-versus-host disease (n = 2), and adenosquamous carcinoma (n = 1). The rate of PGD3 was 9% (n = 5). The 1-year survival rate was 93.4%. CONCLUSION: After initial acceptance, cDCD lung grafts led to LT in 76% of cases, with outcomes similar to those already reported in the literature. The relative impacts of NRP and EVLP on the outcome following cDCD LT should be assessed prospectively in the context of comparative studies.

Double-blind comparison of ketoprofen and placebo in the treatment of sprains and strains.

Sixty patients (42 men and 18 women between the ages of 18 and 37 years) with grade 2 or grade 3 sprains or elongations participated in a seven-day double-blind study. By random determination, they were treated with either 100 mg of ketoprofen or placebo TID. Pain evaluation, vital signs, concomitant medication, and side effects were determined on days 1,3, and 7. Compared with placebo, ketoprofen produced statistically significant reductions (P less than 0.05) in investigator-rated pain at rest (day 3), on palpation (days 1, 3, and 7), on motion (days 1 and 3), and on the subject-rated analog scale (days 1 and 3). It also achieved a greater relief of pain (days 1 and 3) and greater overall efficacy, as expressed by the statistically significant improvements in the total pain scores on motion and on palpation, on the total pain relief score, on the total pain analog score, and on the sum of pain intensity differences for pain at rest, on palpation, and on motion. Tolerance to the seven-day ketoprofen treatment was excellent. Results from the study indicate that ketoprofen is efficacious in relieving pain following sprains and strains.

The acoustic stapedial reflex as a prognostic indicator in sudden onset sensorineural hearing loss.

Forty consecutive patients with sudden onset sensorineural hearing loss treated similarly over a four-year period were studied retrospectively to determine a correlation between presence or absence of the acoustic stapedial reflex (ASR) and recovery of hearing. Six patients were dropped from the study because of incomplete data. There were 42 affected ears in the remaining 34 patients. The ASR was present in 28 (67%) of the 42 affected ears that had either full or partial recovery. The ASR was not present in 11 ears (26%) in which there was no recovery. The ASR was present in three ears (7%) in which there was no recovery. There was a statistically significant association (P < .001) between presence of the ASR and eventual full or partial recovery of hearing in this series. The ASR would have been an effective prognostic indicator for 32 (94%) of our 34 patients (39 [93%] of 42 affected ears).

[Illusion of visual tilt: a case]. / Illusion visuelle d'obliquité: un cas.

A 21-year-old man experienced sudden and intense rotational vertigo. A moment later, he was amazed to see the whole visual world rotate clockwise progressively by about 160 degrees. On the following day, the visual tilt was only 30 degrees, and it soon receded completely, while headache appeared. CT scan, magnetic resonance imaging, vertebral arteriography, and cerebrospinal fluid analysis were normal, and the aetiology could not be ascertained. Illusions of visual tilt are an unusual consequence of disorders affecting the vestibular system. The locus of the impairment is usually in the brainstem, particularly in association with Wallenberg's syndrome. However, the peripheral vestibular pathways or the vestibular projections into the cerebral cortex may occasionally be involved. In the latter case, the visual tilt is part of a vestibular epileptic seizure.

Increased expression of adhesion molecules (ICAM-1 and LFA-1) on alveolar macrophages from asthmatic patients.

In the airways inflammation observed in asthma, activated macrophages are present in increased numbers. Adhesion molecules are required for the cell:cell contacts between leukocytes and endothelial cells or other leukocytes, and they are induced by inflammatory stimuli. We studied the expression of two adhesion molecules (ICAM-1 and LFA-1) on alveolar macrophages recovered by bronchoalveolar lavage from 11 normal subjects and 13 asthmatic patients by using immunocytochemistry. Two specific monoclonal antibodies were used, and the reaction was revealed by the alkaline phosphatase-antialkaline phosphatase (APAAP) method. The percentage of cells expressing ICAM-1 or LFA-1 was significantly increased in asthmatic patients, as compared with normal subjects (P < 0.001 and P < 0.002, respectively; Mann-Whitney U test), and there was a significant correlation with the percentage of cells expressing both markers in asthma (P < 0.03, Spearman rank test). This study highlights the importance of macrophages in the inflammation of asthma and suggests that macrophage interactions with other cells play a role in this inflammation.

Activation by histamine of bronchial epithelial cells from nonasthmatic subjects.

Histamine is a major mediator of the mast cells that are present between epithelial cells in asthma. In asthma, there is an increased expression of ICAM-1 and HLA-DR and an increased spontaneous release of fibronectin. The effect of histamine was tested on bronchial epithelial cells obtained by bronchial brushing from 22 nonasthmatic subjects. The activation of epithelial cells was assessed by immunocytochemical analysis of the expression of membrane markers (ICAM-1 and HLA-DR) using the alkaline phosphatase-anti-alkaline phosphatase method and the release of fibronectin (enzyme immunoassay). Time-response (three experiments) and dose-response (six experiments) curves showed that the maximal effect was obtained after an incubation time of 24 h and a dose of 1 microM of histamine. For this time course and concentration, there was a highly significant increase in the number of cells expressing ICAM-1 (before histamine: 10 +/- 11%; after histamine: 32 +/- 20%; P < 0.001) and HLA-DR (before histamine: 8 +/- 7%; after histamine: 23 +/- 20%; P < 0.001) and in the release of fibronectin (before histamine: 30 +/- 20 ng/10(5) viable cells; after histamine: 61 +/- 35 ng/10(5) viable cells; P < 0.003). Cycloheximide blocked these effects, suggesting that histamine requires protein synthesis for its action. Pyrilamine (H1-blocker) and ranitidine (H2-blocker) at a concentration of 10 microM decreased the effect of histamine. However, there was no additive effect when both antagonists were added. This study suggests that mast cells present in the airways have a role in the activation of epithelial cells.

HLA-DR and ICAM-1 expression on bronchial epithelial cells in asthma and chronic bronchitis.

HLA-DR and ICAM-I molecules play an important role in cellular interactions, and their expression can be induced by inflammatory stimuli. We evaluated the spontaneous expression of HLA-DR and ICAM-1 on epithelial cells obtained by bronchial brushing from 27 asthmatic patients, 10 chronic bronchitis (CB) patients, and 19 normal subjects. In all cases > 90% pure epithelial cells were obtained. HLA-DR and ICAM-1 were characterized using monoclonal antibodies and revealed by immunocytochemistry (APAAP technique). In asthma, the percentage of cells expressing HLA-DR and ICAM-1 was significantly increased by comparison with normal values (p < 0.001) and GB (p < 0.001; HLA-DR; p < 0.003; ICAM-1) and was correlated with the clinical score of Aas (p < 0.001, HLA-DR; p < 0.001, ICAM-1) and the FEV1 (p < 0.001, HLA-DR; p < 0.002; ICAM-1). In CB, expression of both markers was slightly but significantly increased by comparison with normal subjects and was correlated with FEV1 (p < 0.02, HLA-DR; p < 0.03, ICAM-1). In addition, the expression of HLA-DR and ICAM-1 was significantly correlated. This study suggests that epithelial cells are in an activated state in asthma and that the extent of expression of these markers may be specific to asthma, not a general feature of chronic inflammation.

Phenotypic and functional modulation of normal human alveolar macrophages by histamine.

Alveolar macrophages (AM) play a regulatory role in asthma. AM from asthmatics are activated, release increased amounts of cytokines, and express higher levels of the low affinity receptor for IgE (Fc epsilon RIIb/CD23b) and receptors for adhesion molecules. The bronchial microenvironment may modulate the phenotypic and functional characteristics of AM. On AM from normal subjects, the effects of histamine were studied on the expression of adhesion molecules (LFA-1, ICAM-1) and CD23b as well as on the release of fibronectin. The expression of LFA-1, ICAM-1, and CD23b was examined by immunocytochemistry using the alkaline phosphatase-anti-alkaline phosphatase technique. The expression of CD23b mRNA was studied by in situ hybridization. The release of fibronectin was measured by enzyme immunoassay. We found that histamine induced in a dose- and time-dependent fashion a significant increase of AM expressing the three membrane markers and a significant increase in the release of fibronectin. The maximum effect of histamine was observed after an incubation of 12 to 24 h and a dose of 1 microM. The histamine effects were specific, since they were significantly inhibited by an H1-blocker, pyrilamine, used at a concentration of 10 microM. The effect of an H2-blocker (ranitidine, concentration of 10 microM) was inconstant. Cycloheximide blocked the histamine effects, suggesting that it requires protein synthesis for its action. This study provides an in vitro model of cellular interaction between mast cells and AM, which might be relevant in the airway inflammation in asthma.