The induction of cardiac ruptures by HSA-Flag-TWEAK depends on neutrophils.

<p>Mice were treated with an anti-Ly6G antibody to deplete neutrophils (A) in the myocardium as measured by immunohistological staining and (B) in the peripheral blood as measured by FACS analysis. (C) Neutrophil depletion did not affect survival after MI in PBS and HSA-Flag-TWEAK treated mice. (D) The occurrence of cardiac ruptures was significantly reduced after anti-Ly6G antibody treatment in the HSA-Flag-TWEAK treated group in comparison to the neutrophil intact control.</p

HSA-Flag-TWEAK increases immune cell infiltration into the infarcted heart.

<p>(A) Exogenous administration of HSA-Flag-TWEAK amplified the infiltration of leukocytes into the infarcted myocardium 3 days after MI as determined by FACS analysis. (B) Neutrophils highly infiltrated the border zone of HSA-Flag-TWEAK treated mice.</p

HSA-Flag-TWEAK treatment modulates the expression of cytokines and chemokines via NFκB and JAK/STAT-signalling.

<p>The protein-protein interaction network maps cytokine and chemokine data from the protein microarrays onto the human interactome. Circles indicate proteins; kinases are depicted in triangular shape, each specified by gene names. They are connected either by gray lines indicating protein-protein interactions or red arrows denoting phosphorylation reactions. The functional entities are highlighted (interleukins, chemokines, JAK/STAT pathway). Green-colored nodes show up-regulation in the TWEAK stimulation (orange) condition.</p

HSA-Flag-TWEAK modulates the production of cytokines in the infarcted heart after MI.

<p>u.d. = under detection limit.</p><p>IFN-γ: <11 pg/ml; IL-12: <5 pg/ml; MCP-1: <5 pg/ml; MCP-5: <1 pg/ml; RANTES: <3 pg/ml.</p><p>A cytokine array for the detection of the protein expression of different cytokines was performed 3 days after MI. HSA-Flag-TWEAK treated mice showed statistically significant up-regulation of interleukin 12 (IL-12), interleukin 5 (IL-5), monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-2 (MIP-2) and regulated on activation, normal T cell expressed and secreted (RANTES) (u.d. = under limit of detection).</p

HSA-Flag-TWEAK does not induce cardiomyocyte apoptosis.

<p>The percentage of apoptotic cells 3(A) TUNEL-positive cells and (B) cleaved PARP was similar between HSA-Flag-TWEAK and PBS treated mice.</p

HSA-Flag-TWEAK fails to modulate extracellular matrix remodeling after MI.

<p>mRNA-expression of (A) collagen1α1, collagen1α2, (B) MMP-2, MMP-3, MMP-8, and MMP-9 were unaffected in the scar region of HSA-Flag-TWEAK challenged mice as were the (C) zymographic activities of MMP-2 and MMP-9 (measured as gel band intensity) and (D) TIMP-2, TIMP-3, and VEGF mRNA expression.</p

HSA-Flag-TWEAK does not affect echocardiographic measurements after MI.

<p>Animals underwent echocardiography on day 1, day 3, day 21 (data not shown) and day 56 (data not shown) after MI. All measurements were recorded at the midpapillary level which shows changes in the dimensions of the surviving non-infarcted myocardium, as well as on the apical level depicting changes in scar formation.</p><p>Data are means ± sem; <i>n</i> indicates number of animals studied. EDA, end-diastolic area; ESA, end-systolic area; FS, fractional shortening; 2D, 2-dimensional.</p

HSA-Flag-TWEAK increases mortality and cardiac ruptures after MI.

<p>(A) Percent survival was significantly deceased in HSA-Flag-TWEAK treated mice after MI compared to placebo-treated mice. (B) Most HSA-Flag-TWEAK treated mice died because of left ventricular ruptures.</p

MI induces the expression of TWEAK and Fn14 in the mouse heart.

<p>(A) TWEAK and (B) Fn14 mRNA expression were significantly increased 3 days after experimental MI. Fn14 mRNA expression was significantly increased after 8 weeks of MI. (C) Immunohistological staining after 3 day of MI revealed high expression of Fn14 protein in the border zone of the myocardium co-localizing with periostin expression. (D) Periostin positive fibroblasts are the main source of Fn14 expression in the infarcted heart and αSAA positive cardiomyocytes show no expression of Fn14. (E) Isolated cardiac mouse fibroblasts express Fn14 whereas (F) isolated cardiomyocytes show no Fn14 expression.</p

Loss of host TNFR1 perturbs the immunologic control of Panc02 tumors.

<p>Panc02-tumors were explanted one month after tumor cell inoculation, sectioned, and stained for different immune cells and blood vessels (B6.WT n = 4, B6.TNF KO n = 5, B6.TNFR1 KO n = 5, B6.TNFR2 KO n = 6, B6.TNFR1R2 KO n = 4).</p>*<p>p≤0.05, ** p≤0.01.</p