Expression of LAIR-1 and NKp44 on peripheral blood pDCs from healthy donors.

<p>Expression of LAIR-1 was assessed on pDCs from healthy donors PBMCs (A) and on purified pDCs (B) following culture in the presence of IL-3 (20 ng/ml), CpG (5 µg/ml) or IFNα (1000 U/ml) for 48 hours. (B): Dotted lines depict unstimulated cells; bold lines depict cells cultured as indicated. Values represent expression Geo-MFI. All experiments related to the analysis of LAIR-1 expression on pDCs cultured with the indicated stimuli for 24 and 48 hours are summarized in (C). Values depict Geo-MFI of positive cells/Geo-MFI of Ig isotype-matched control stained cells. (D): Analysis of LAIR-1, NKp44 and DAP12 mRNA expression on human pDCs. Polyclonal NK cell lines, pDCs and pDCs cultured 48 h with IL-3 were analyzed by RT-PCR for LAIR-1, NKp44 and DAP12 expression (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0015080#s4" target="_blank">Materials and Methods</a>).</p

Expression of LAIR-1 and NKp44 on peripheral tissue pDCs.

<p>pDCs were identified in different tissues as BDCA-2⁺ ILT3⁺ positive cells (A). LAIR-1 and NKp44 expression was evaluated in tonsils (B), lymph nodes (C and E) and in different malignant tumors (D and E). pDCs activation in lymph nodes and in NSCLC was assessed as CD83 expression (E and F). The horizontal bars within dot-plots of panels C, D, F represent negative controls with isotype-matched irrelevant mAbs. Data shown are representative of at least three independent experiments analyzing pDCs from different patient tissues.</p

LAIR-1 triggering on pDCs induces inhibition of IFNα production.

<p>LAIR-1 cross-linking inhibits IFNα production by pDCs activated with either CpG (A) or DNA immunocomplexes (B). pDCs were cultured for 18 h in the presence of IL-3, incubated with the indicated mAbs or isotype-matched mAbs (Ctrl mAb) and then cultured in F(ab)2 goat anti-mouse IgG antibody-coated plates in the presence of CpG or DNA immunocomplexes. IFNα production was assessed by ELISA in culture supernatants. Graphs represent mean values +/− SEM of three independent experiments. ** p<0.01; * p<0.05.</p

Allelic and genotype frequencies given as %(n) in German cases compared with age-, gender- and ethnicity matched controls.

*<p><i>P</i><0.001, OR: 0.20, 95%CI: 0.09–0.14.</p>**<p><i>P</i> = 0.001, OR: 0.60, 95%CI: 0.44–0.81.</p>***<p><i>P</i> = 0.0002, OR: 0.35, 95%CI: 0.20–0.61.</p>****<p><i>P</i><0.001, OR: 0.28, 95%CI: 0.17–0.44.</p>*****<p><i>P</i><0.001, OR: 0.63, 95%CI: 0.49–0.80.</p>°<p><i>P</i> = 0.0003, OR: 0.37, 95%CI: 0.22–0.63.</p>°°<p><i>P</i> = 0.01, OR: 0.66, 95%CI: 0.49–0.90.</p>°°°<p><i>P</i> = 0.002, OR: 0.48, 95%CI: 0.31–0.77.</p>°°°°<p><i>P</i><0.001, OR: 0.43, 95%CI: 0.30–0.63.</p>°°°°°<p><i>P</i> = 0.005, OR: 0.70, 95%CI: 0.55–0.89.</p>•<p><i>P</i> = 0.01, OR: 0.51, 95%CI: 0.30–0.85.</p>••<p><i>P</i><0.0001, OR: 0.53, 95%CI: 0.39–0.72.</p>•••<p><i>P</i> = 0.019, OR: 0.62, 95%CI: 0.42–0.91.</p>••••<p><i>P</i> = 0.007, OR: 0.71, 95%CI: 0.56–0.91.</p

Characteristics of Italian (I) and German (G) subjects included in plasma level evaluation.

<p>Characteristics of Italian (I) and German (G) subjects included in plasma level evaluation.</p

Characteristics of German individuals included in the association study.

*<p> <b>age at disease onset.</b></p

Scattergram of progranulin plasma levels in BPD patients and controls.

<p>Black lines represent mean values. G = German. I = Italian. *<i>P</i><0.001, patients versus controls.</p