Abstracts from CIPP XVI Meeting Libon june 2017.pdf

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and increased BSM mass. Preschool children with increased

A transversal study was performed, involving 40 pediatric

BSM mass are at a greater risk of developing asthma

patients, 20 with PIBO and 20 with TRSA. Pulmonary function


was performed with a KoKo spirometer. Bronchial provocation

increased mitochondrial content leading to greater BSM cell

was performed with inhalation of carbachol until achieving a drop


of 20% in FEV1. ENO was performed with Nioxmino


in a

registered exhaled stream of 0,35L/s. Induced sputum cellularity
was obtained using a 4.5% hypertonic saline solution or 0.9%









Mitochondria may represent a new target for treating BSM proliferation in severe childhood asthma.

physiological solution in stable patients and with FEV1 values
after bronchodilator higher or equal to 60% of predicted or lower


than 60% respectively, during four periods of five minutes each,

To investigate in vitro BSM cell proliferation in severe asthmatic

totaling a maximum time of 20 minutes of inhalation. Alterations

children. Moreover, we sought to determine the effect of

in tomography were analyzed through a score punctuation and by

conventional and non-conventional anti-asthma treatments on

the presence of tomographic alterations. ROC curves were

BSM proliferation and verify whether the underlying mechanisms

performed to evaluate which variable could discriminate these

involve mitochondria.

two diseases. Results: The patients with PIBO had lower values of


FEV, FEF25-75% and FEV1/FVC and total lung capacity (TLC).
The most frequent tomographic alterations in POBI were:
bronchiectasis (90%), air trapping (90%) and mosaic attenuation
(85%), all with statistical significance comparatively to TRSA. In
ROC curves, an area under curve (AUC) higher or equal to 0,8 was
observed for the following variables: Blomia tropicalis, ENO,
tomographic score, severity of bronchiectasis, generation of
bronchial division, mosaic attenuation, air trapping, FEF25-75%,
FEV1/FVC, variation in FEV1 after oral corticosteroids and the
association of mosaic pattern and ENO. In this study, no
parameter of atopic markers reached a sensitivity and specificity
higher than 80% while, for pulmonary function parameters, only
FEF25-75% and FEV1/FVC reached these levels. However, only
the tomographic score and mosaic attenuation showed a
sensitivity and specificity higher or equal to 95. Only two studies
were found in the literature comparing PIBO and TRSA both of
which involved mixed samples including children, teenager and

BSM cells were cultured from bronchial biopsies obtained
from severe asthmatic preschool children (1 to 4 years old).
BSM cell proliferation was assessed by manual counting
and flow cytometry (CFSE, CellTrace proliferation kit) after
5 days in culture medium containing 10% fetal calf serum
(10% FCS). Cells were then treated with Dexamethasone,





Verapamil or Azithromycine (10–9 to 10–6M for all agents) in
the presence of 10% FCS. Cell viability was assessed using Trypan
blue staining solution and flow cytometry after diamidinophenylindol (DAPI) staining. Cellular cycle and apoptosis were
assessed after DAPI and Annexin-PI staining, respectively.
Mitochondrial mass, biogenesis and autophagia were assessed
by Western blot and Flow Cytometry, using anti-Porin,
anti-mitochondrial transcription factor A and anti-LC3A/B
antibody, respectively.

adults. In one of the studies, a statistically significant difference


was only observed in mosaic attenuation, while in the other, a

Cells were attributed to 2 groups according to muscle

statistically significant difference was observed in mosaic

mass on histochemical analysis: big and small BSM ((BM and

attenuation, air trapping and bronchiectasis.

SM respectively) based on the Z-score determined from normal


samples. The mean + SEM number of cells in the BM group

The presence of higher scores in tomography and the presence
of mosaic attenuation were found to best differentiate TRSA and

significantly increased after 5 days of culture vs. the SM group
186 700 + 35 800 vs. 61 560 + 11 900 BSM cells, respectively,
p < 0.001). Only Azithromycin (10–7M) significantly decreased
BSM cell proliferation by 1/3 without increasing the number of
dead or apoptotic cells or blocking the cell cycle. Azithromycin
decreased mitochondrial mass by 1/4 in particular in BM cells, by

#A26 - Azithromycin Decreases in vitro Bronchial Smooth

increasing autophagia but had no influence on mitochondrial

Muscle (BSM) Cell Proliferation in Severe Pediatric Asthma.


Beaufils F., Siao V., Trian T., Berger P., Fayon M.


Centre de Recherche Cardio-thoracique de Bordeaux. INSERM UI045 –
Bordeaux, France

BSM cells from severe asthmatic children show varying


degrees of proliferation. Increased BSM mass is an indicator
of increased mitochondrial content in BSM cells. Azithromycin


Severe asthma in childhood is associated with decreased

and decreasing BSM cell proliferation, particularly in the BM

lung function in adulthood. This is linked to airway remodeling