Characteristics of the cystic fibrosis patients.

<p>Characteristics of the cystic fibrosis patients.</p

Individual values of inspiratory resistances before and after autogenic drainage in 30 CF patients.

<p>A, Rrs_5; B, Rrs_11; C, Rrs_19; D, Rrs₅–Rrs₁₉.</p

Polycyclic Aromatic Hydrocarbons Reciprocally Regulate IL-22 and IL-17 Cytokines in Peripheral Blood Mononuclear Cells from Both Healthy and Asthmatic Subjects

<div><p>Pollution, including polycyclic aromatic hydrocarbons (PAH), may contribute to increased prevalence of asthma. PAH can bind to the Aryl hydrocarbon Receptor (AhR), a transcription factor involved in Th17/Th22 type polarization. These cells produce IL17A and IL-22, which allow neutrophil recruitment, airway smooth muscle proliferation and tissue repair and remodeling. Increased IL-17 and IL-22 productions have been associated with asthma. We hypothesized that PAH might affect, through their effects on AhR, IL-17 and IL-22 production in allergic asthmatics. Activated peripheral blood mononuclear cells (PBMCs) from 16 nonallergic nonasthmatic (NA) and 16 intermittent allergic asthmatic (AA) subjects were incubated with PAH, and IL-17 and IL-22 productions were assessed. At baseline, activated PBMCs from AA exhibited an increased IL-17/IL-22 profile compared with NA subjects. Diesel exhaust particle (DEP)-PAH and Benzo[a]Pyrene (B[a]P) stimulation further increased IL-22 but decreased IL-17A production in both groups. The PAH-induced IL-22 levels in asthmatic patients were significantly higher than in healthy subjects. Among PBMCs, PAH-induced IL-22 expression originated principally from single IL-22- but not from IL-17- expressing CD4 T cells. The Th17 transcription factors <i>RORA</i> and <i>RORC</i> were down regulated, whereas AhR target gene <i>CYP1A1</i> was upregulated. IL-22 induction by DEP-PAH was mainly dependent upon AhR whereas IL-22 induction by B[a]P was dependent upon activation of PI3K and JNK. Altogether, these data suggest that DEP-PAH and B[a]P may contribute to increased IL22 production in both healthy and asthmatic subjects through mechanisms involving both AhR -dependent and -independent pathways.</p></div

Cytokine profile and cell expression of PAH-stimulated peripheral blood mononuclear cells cultured in Th17 conditions.

<p><b>A</b> IL-17A and IL-22 production by activated peripheral blood mononuclear cells from 6 nonallergic (NA) subjects and 6 allergic asthmatic (AA) patients stimulated or not with SRM or B[a]P for 6 days was determined by ELISA. Results are expressed as mean ± SEM. *<i>P</i><.05 versus corresponding control. ^#<i>P</i><.05 AA versus NA subjects. <b>B</b> Activated peripheral blood mononuclear cells were stimulated or not with SRM and B[a]P, stained with antibodies against cytokine and cell surface markers, and evaluated by flow cytometry. Results are expressed as mean ± SEM percentage of cytokine positive cell populations for n = 7–11 subjects. *<i>P</i><.05 versus corresponding control, ^##<i>P</i><.01, ^###<i>P</i><.001.</p

Transcript levels of genes involved in Th17/Th22 polarization.

<p>Activated PBMCs from nonallergic (NA) subjects (n = 8) and allergic asthmatic (AA) patients (n = 11) were incubated with or without PAH for 72hr, and gene mRNA level was assessed by Q-RT-PCR. Results are expressed as mean relative expression (RE) of 2^(-ΔCt) ± SEM, where the ΔCt value of the sample was determined by subtracting the Ct value of the target gene from the Ct value of the rs9 house keeping gene. *<i>P</i><.05,**<i>P</i><.01, ^#<i>P</i><.05 and ^##<i>P</i> <.01 AA versus NA subjects.</p

Effects of AhR antagonist on IL-22 and IL-17A secretion by peripheral blood mononuclear cells from Allergic asthmatics (AA) and nonallergic subjects (NA).

<p>Activated peripheral blood mononuclear cells from NA subjects (n = 10) and AA patients (n = 12) were incubated or not with PAH, in the presence or not of AhR antagonist CH-223191 for 72hr, and cytokine production was assessed by ELISA in the supernatants. Results are expressed as mean ± SEM. *<i>P</i><.05, **<i>P</i><.01. The dotted line is set on the level of the antagonist-treated control cells.</p

Cytokine profile of PAH-stimulated peripheral blood mononuclear cells in allergic asthmatics (AA) and nonallergic subjects (NA).

<p>IL-17A, IL-22 and IL-10 secretion by activated peripheral blood mononuclear cells from NA subjects (n = 10) and AA patients (n = 12) stimulated or not with different PAH for 72hr was determined by ELISA. Results are expressed as mean ± SEM. *<i>P</i><.05 and **<i>P</i><.01 versus control. ^#<i>P</i><.05 and ^##<i>P</i><.01 AA versus NA subjects.</p

Number of 16S-pyrosequencing reads assigned to each taxonomic group of Bacteria.

a<p>Once a read was assigned to the highest taxonomical level according to the criteria defined in material and method section, it was not added up in the next taxonomic level.</p

Relation between species richness and clinical status (A) or lung function (B).

<p>Total richness of prokaryotic and fungal communities from each patient-sample was expressed using the Chao1 richness estimator; each spot size is proportional to the corresponding Chao1 value. The clinical status is expressed as S-K score and BMI in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036313#pone-0036313-g002" target="_blank">Figure 2A</a>, while lung function is expressed as FEV1 and FVC values in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036313#pone-0036313-g002" target="_blank">Figure 2B</a>. Given to the absence of S-K score value from Patient 2-sample 2 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036313#pone-0036313-t001" target="_blank">Table 1</a>), this spot is missing in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036313#pone-0036313-g002" target="_blank">Figure 2A</a>. Bacterial and fungal Chao1 values corresponding to Patient 1, Patient 2, Patient 3, and Patient 4 are represented in blue-, green-, red- and yellow-edged spots, respectively. Dark and light colour intensity is corresponding to the first and second sampling dates of each patient, respectively. Dark grey and light grey are corresponding to fungal and bacterial Chao1 richness values, respectively.</p

Clinical data and treatment from CF patients included in the study.

a<p>S-K score, Shwachman-Kulczycki Score;</p>b<p>BMI, body mass index;</p>c<p>FVC, forced vital capacity;</p>d<p>FEV1, forced expiratory volume;</p>e<p>ATB, antibiotic drug;</p>f<p>ATF, antifungal drug;</p>g<p>ITC, itraconazole;</p>h<p>UNK, unknown CFTR mutations associated with an abnormally high sweat chloride test (110 mmol/L).</p