Overlap of serum markers correlating with outcome of islet transplantation.

<p>Venn diagram showing serum markers that predict or correlate with clinical outcome (good or poor graft function) 1 year after islet transplantation.</p

Serum marker levels before or after islet transplantation that could differentiate between good and poor graft function.

<p>Heatmap representation of significantly different serum markers between patients with good graft function (green) and patients with poor graft function (orange) before (A&B) or 1 year after (C&D) transplantation (Student’s t-test, p<0.05). In the group of patients with good graft function, those with continued insulin independence (green) and temporary insulin independence (grey) were subdivided (B & D). Heatmap gradient represents min to max (cyan—black—red) normalized serum titers.</p

Serum markers changing by transplantation and immunosuppression.

<p>Serum marker levels that significantly change from pre islet cell transplantation to one year post transplantation are depicted (Student’s t-test, p<0.05): for all patients (A), for patients with good engraftment (B, green) and for patients with insufficient engraftment (C, orange). Serum levels are in pg/ml for: IL-7, IL-11, IL-13, IL-15, IL-16, IL-22, LIF, TSLP, CCL2, FAS, BDNF, KIM-1; ng/ml for: IL-23, CCL22, Cathepsin S, sICAM, S100A12; in ug/ml for: Chemerin, Leptin; mg/ml for: Adiponectin. Samples out-of-range at both time points were excluded from statistical analysis.</p

Adiponectin receptor 1 expression.

<p>Adiponectin receptor 1 expression.</p

Adiponectin receptor 2 expression.

<p>Adiponectin receptor 2 expression.</p

Leukocyte subset numbers.

<p>Leukocyte subset numbers.</p

Patient characteristics.

<p>Patient characteristics.</p

IL-7 and TLR7 synergistically increases CD4 T cell proliferation in T and B co-cultures.

<p>Representative FACS stainings for KI67⁺ CD4 T cells from unstimulated, IL-7, and TLR7/IL-7 stimulated T/B cell co-cultures in the absence or presence of monocytes/macrophages are shown (n = 5). IL-7 significantly stimulates proliferation of CD4 T cells, which is synergistically increased when combined with TLR7 stimulation (<b>A, B</b>). IL-7-induced CD4 T cell proliferation is enhanced in the presence of monocytes/macrophages, but no additive effect is observed with a combination of IL-7 and TLR7 stimulation (<b>C, D</b>).*p<0.05 and **p<0.001 indicate statistical significance compared to medium values.</p

Adipokine receptor expression on leukocyte subsets.

<p>The first column illustrates the gating strategy for the leukocyte subsets. The other columns show representative histograms of the AdipoR1, AdipoR2 and leptin receptor expression of the different leukocyte subsets.</p

IL-7 synergistically increases proliferation of TLR7-stimulated B cells in co-culture with CD4 T cells, which is enhanced by monocytes/macrophages.

<p>Isolated B cells co-cultured 1∶1 (5.10⁵ each) together with CD4 T cells for 6 days show an increased lymphocytic proliferation upon TLR7 or IL-7 stimulation, which is additively increased upon combined stimulation with IL-7/TLR7 (n = 8) (A). A similar, but overall enhanced effect for the total proliferation is seen when monocytes/macrophages are added to the culture (5.10⁴ B). Representative FACS stainings for KI67⁺ B cells from an unstimulated, TLR7, IL-7, and TLR7/IL-7 stimulated CD4 T/B cell co-culture -/+ monocytes/macrophages are shown as well as the average data (n = 5) (C, D). TLR7 induces a significant increase in the percentage of KI67⁺ B cells. IL-7 stimulation induces a small, but statistically significant increase in Ki67⁺ B cells. When TLR7 and IL-7 are added together a synergistic increase in proliferation is observed (C). Overall the effects are enhanced by addition of monocytes/macrophages to the T/B cell co-cultures (D). * and ** indicate a statistical significant differences of p<0.05 and p<0.01, respectively, as compared to medium values or between treatments.</p