Influence of CLL18 on chemoresistance.

<p>Cells were cultured in presence or absence of -TGFbeta or CCL18 in the concentrations indicated for 72 h. After the culture period, medium was changed and the cells were incubated with cisplatin for additional 72 hours at concentrations ranging from 0.4 to 25 µg/ml. Cell survival was measured by MTT assay. The horizontal line indicates the LD₅₀, vertical lines indicate the LD₅₀ for the respective pre-incubation condition (n = 4).</p

Blocking of CD204 abrogates effects mediated by collagen type I monomers.

<p>Preincubation of AM from HD (n=10, Panel A) and IPF patients (n=8, Panel B) with neutralizing antiCD204 (2µg/ml) prior to cell culture abrogated collagen type I monomers induced increase in CCL18 production. Mouse IgG1 served as control. Data are expressed by mean ± SD (*, <i>p</i><0.05).</p

CCL18 suppreses proliferation of A549 cells. (n = 7 in each group).

<p>CCL18 suppreses proliferation of A549 cells. (n = 7 in each group).</p

mRNA levels of surfactant proteins in human AEC-II.

<p>SP-A1, SP-A2, SP-B, SP-C and SP-D mRNA expression level of human AEC-II (freshly isolated or cultured for 24 h) were measured by real-time PCR. Bar charts show mean ± SD (n = 5, native = freshly isolated non-cultured cells; 24 h culture  =  isolated cells cultured as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038369#s2" target="_blank">Material and Methods</a>).</p

Collagen type I monomers increase Phospho-Akt expression via PI3kinase activation.

<p>Stimulation of AM with collagen type I monomers resulted in a moderate increase in phospho-Akt expression, which was more pronounced in IPF patients (Panel <b>A</b>). Basal, constitutive phospho-Akt expression was high in AM from each 5 healthy donors and IPF patients. A non-selective inhibitor of PI3kinase, LY294002 (50 μM, DMSO as control), abrogated CCL18 production following stimulation with collagen type I monomers and decreased significantly spontaneous CCL18 production of AM (Panel <b>B</b>). Data are expressed by mean ± SD (*, <i>p</i><0.05; **, p<0.005).</p

Figure 1

<p>Morphological changes in cells treated with CCL18 and TGFbeta: untreated A549 cells show typical epithelial morphology with close cell-cell-contacts, which are marked by white arrows (A). After treatment with 2 ng/ml -β cells changed the phenotype to a more mesenchymal morphology with prolonged cell extrusions and loosened cell-cell-contacts marked by arrowheads. (B) CCL18 also changes the cell morphology in a dose dependent manner (C [1 ng/ml] and D [10 ng/ml]) to fibroblast-like mesenchymal phenotype and loosened cell-cell-contacts marked by arrowheads.</p

Collagen monomers increase M2 marker production by AM.

<p>M2 cytokine production of BAL cells stimulated with monomers of various collagen types. M2 cytokine production of BAL cells from 5 additional patients with IPF are depicted in black and 5 additional experiments with BAL cells from healthy donors are depicted in grey. <b>Panel A</b> shows CCL18 production following stimulation with rat collagen-I (Col 1R; 100µg/ml), human collagen-I (Col I, 100 µg/ml), human collagen-III (Col III, 100 µg/ml), human collagen-IV (Col IV, 100 µg/ml), human collagen-V (Col V, 100 µg/ml). <b>Panel B</b> shows IL-1ra production following the same stimulation protocol. <b>Panel C</b> shows CCL2 production. *p<0.05.</p

Estimated effects from univariate Cox regression models for acute exacerbations.

<p><b>Definition of abbreviations:</b> FVC: forced vital capacity; n.s.: not significant.</p><p>Estimated effects from univariate Cox regression models for acute exacerbations.</p

Spontaneous macrophage derived chemokine production in IPF patients w/o acute exacerbation.

<p>Boxplots of the spontaneous production of A: CCL2, B: CCL17, C: CCL18, D: CCL22, E: IL-1ra, F: TNF-α, G: IL-1β, H: IL-8 and I: CXCL1 protein by BAL cells of patients with IPF. The dark grey boxplots represent patients who suffered from an acute exacerbation (AE, n = 12), the light grey represent patients who did’t suffer from an AE at timepoint of BAL (NoAE, n = 59) (* p<0.05, ** p<0.01, n.s. = not significant).</p

CCL18 serum level and survival of patients with NSCLC in days from time point of diagnosis.

<p>Survival time varies significantly with different serum CCL18 levels (p<0.004).</p