NEMO deficiency in recipient mice favours donor cell selection.

<p>(<b>A</b>) Hepatocyte transplantation was applied to NEMO^Δhepa and WT recipient mice with preceding BMT. Donor cells were derived from WT (Casp8^loxP/loxP/hAAT(+)) or Caspase-8-deficient (Casp8^Δhepa/hAAT(+)) mice, respectively. (<b>B</b>) Serum hAAT levels were analysed via quantitative ELISA and displayed on a logarithmic scale. Basic values represent serum hAAT level 1d post transplantation and are used as a reference value for the calculation of the relative increase in the amount of donor derived hepatocytes over time. (<b>C</b>) Quantitative evaluation of hAAT (+) liver tissue areas 52 weeks after HT. (<b>D</b>) Visualization of engrafting donor cells using hAAT immunofluorescence staining. Clusters of donor derived hepatocytes are displayed 12 and 52 weeks after HT (blue: DAPI; red: hAAT(+) cells).</p

Quantitative histological assessment of mouse livers.

<p>Liver histologies were graded for fibrosis levels according to an adapted METAVIR fibrosis score. F0 =  no fibrosis; F1 =  portal fibrosis without septa; F2 =  portal fibrosis with septa; F3 =  numerous septa without cirrhosis; F4 =  cirrhosis.</p

Reduced fibrogenesis in hepatocyte transplanted NEMO^Δhepa mice.

<p>(<b>A</b>) Analysis of Collagen accumulation in transplanted mice via Sirius red staining under polarized light. NEMO^Δhepa mice that underwent transplantation with either Casp8^loxP/loxP/hAAT(+) or Casp8^Δhepa/hAAT(+) donor cells 52 weeks after HT were compared to age-matched completely untreated control mice as well as to bone marrow-transplanted mice 56 weeks (age matched) after BMT. (<b>B</b>) Collagen-1α staining for visualisation of a specific fibrotic collagen subtype (blue: DAPI: red/yellow: Collagen1-α). (<b>C</b>) Assessment of Collagen-1α mRNA expression via quantitative realtime PCR. (**p<0.01, ***p<0.001)</p

Assessment of NK cell activation and hepatic proliferation.

<p>(<b>A</b>) NK cell staining was performed in order to address the role of the immune cell response to HT (blue: DAPI; green: NK1.1(+) cells). This shows, that NK-cells are present in a significantly lower number in HT-recipient mice as compared to controls and solely BM-treated animals. (<b>B</b>) Visualization of cell proliferation by Ki-67 and BrdU incorporation stainings. (blue: DAPI; green: BrdU(+); red: Ki-67(+)). (<b>C</b>) Determination of the total number of BrdU(+) cells by means of BrdU staining in a 100x magnification. (*p<0.05, **p<0.01). (<b>D</b>) Quantification of Ki-67 positive signals in immunofluorescent staining using a 200x magnification. (*p<0.05, **p<0.01). (<b>E</b>) For localization of proliferating cells, double immunofluorescence staining of BrdU and hAAT was performed (blue: DAPI; red: hAAT(+) cells; green: BrdU(+) cells). (<b>F</b>) DAB-staining of NEMO protein in NEMO^Δhepa mice transplanted with Casp8^loxP/loxP/hAAT(+) or Casp8^Δhepa/hAAT(+) donor cells, respectively (DAB in brown: NEMO protein). This clearly shows the presence of NEMO(+) hepatocytes in the surrounding NEMO-deficient (NEMO^Δhepa) liver tissue.</p

Anti-food and anti-microbial serum IgG and IgA levels in CD patients with or without stricturing/penetrating disease and controls.

<p>Specific serum IgG (A) and IgA (B) were quantified by ELISA in control patients/healthy controls (n = 61) and CD patients without (n = 17) and with (n = 34) stricturing and/or penetrating complications. Boxes indicate median and 25/75 percentiles and whiskers indicate 10/90 percentiles. P values were determined by Kruskal-Wallis test followed by a Dunn's post hoc test (*<0.05; **<0.01; ***<0.001) or by Mann Whitney U test (# <0.05). Mann Whitney U test was only applied between CD subgroups and results are only shown if the Dunn's post hoc test did not show any significance.</p

Summary of results. Ab levels compared to controls.

1<p>Slightly increased anti-food IgA levels in CD patients with a structuring/penetrating phenotype.</p>2<p>Decreased anti-food IgG levels in CD and UC patients with artopathy.</p>3<p>Only some microbial antigens.</p>4<p>Patients receiving anti-TNFα treatment have lower Ab levels.</p><p>Summary of results. Ab levels compared to controls.</p

Anti-food and anti-microbial serum IgG and IgA levels in CD patients with or without anti-TNFα treatment and controls.

<p>Specific serum IgG (A) and IgA (B) were quantified by ELISA in control patients/healthy controls (n = 61) as well as in CD patients without (n = 17) and with (n = 34) current anti-TNFα treatment. Boxes indicate median and 25/75 percentiles and whiskers indicate 10/90 percentiles. P values were determined by Kruskal-Wallis test followed by a Dunn's post hoc test (*<0.05; **<0.01; ***<0.001) or by Mann Whitney U test (# <0.05). Mann Whitney U test was only applied between CD subgroups and results are only shown if the Dunn's post hoc test did not show any significance.</p

Anti-food and anti-microbial serum IgG and IgA levels in CD patients with or without arthropathy and controls.

<p>Specific serum IgG (A) and IgA (B) were quantified by ELISA in control patients/healthy controls (n = 61) and CD patients without (n = 39) and with (n = 12) current arthropathy. Boxes indicate median and 25/75 percentiles and whiskers indicate 10/90 percentiles. P values were determined by Kruskal-Wallis test followed by a Dunn's post hoc test (*<0.05; **<0.01; ***<0.001).</p

Serum IgG and IgA levels specific for food and microbial antigens in IBD patients and controls.

<p>Serum IgG (A–F) and IgA (G–L) specific for ovalbumin (A/G), wheat (B/H), milk (C/I), mannan from <i>S. cerevisiae</i>  =  ASCA (D/J), and lysates from <i>E. coli</i> K12 (E/K) and <i>B. fragilis</i> ATCC 25285 (F/L) were quantified by ELISA in control patients/healthy controls (Con; n = 61) and patients suffering from CD (n = 52), UC (n = 29) and acute gastroenteritis/colitis (AGE; n = 12). Each dot represents one patient. Medians with interquartile ranges are indicated. P values (*<0.05; **<0.01; ***<0.001) were determined by Kruskal-Wallis test followed by a Dunn's post hoc test.</p