recombinant human vegf c c156s  (R&D Systems)

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Lymphatic and Immune Cell Cross-Talk Regulates Cardiac Recovery After Experimental Myocardial Infarction

doi: 10.1161/ATVBAHA.120.314370

Figure Lengend Snippet: Adeno-associated viral (AAV)-VEGF (vascular endothelial growth factor)-CC156S therapy stimulates cardiac lymphangiogenesis postmyocardial infarction (MI). Mice were treated with AAV-hVEGF (human VEGF)-C C156S gene therapy (red circles, n=8), sVEGFR3 (vascular endothelial growth factor receptor 3; AAV-sVEGFR3, blue triangles, n=5–7), or AAV-scrambled virus (MI controls, black circles, n=8), and sham-operated mice (white circles, n=7–9) served as healthy controls. Cardiac gene expression of lymphangiogenic factors at 21 days post-MI ( A ). Lymphangiogenesis in the viable left ventricular (LV) wall bordering the infarct was evaluated as % proliferating lymphatic vessels ( B ), lymphatic densities ( C ), and lymphatic area at 7 or 21 days post-MI ( D ). Examples of light sheet imaging ( E , left ) of Lyve1 + (lymphatic vessel endothelial receptor 1) cardiac lymphatics in healthy mice, in AAV-MI controls, and VEGFC-treated (AAV–VEGF-C C156S ) mice at 21 d post-MI. Infarct outlined by white dashed lines. Scale bar=1 mm; ×0.8. Confocal imaging ( E , right ) of viable LV areas in the same samples. Cardiac precollectors indicated by white asterisk. Scale bar=50 µm; ×25. Relative cardiac expression of lymphatic-related genes in sham (n=8), MI controls (AAV-scrambled virus, n=8), and VEGFC-treated (AAV-hVEGF-C C156S , n=5) at 21 days post-MI ( F ). Kruskal-Wallis followed by Dunn posthoc test. * P <0.05 vs sham; # P <0.05; ## P <0.01; ### P <0.001 vs AAV-MI controls. con indicates control.

Article Snippet: Modulation of cardiac lymphangiogenesis was performed using either systemic growth factor therapy with recombinant human VEGF-C C156S protein (RnD system) as described, or using viral gene vectors encoding hVEGF-C C156S (human VEGF-C C156S ) or sVEGFR3 (soluble VEGFR3)-IgG construct., , Briefly, protein therapy consisted of repeated (day 0, 2, 3, 4, and 6 post-MI) intraperitoneal injections of 2 µg/mouse (0.1 μg/g) of rhVEGF-C C156S (recombinant human VEGF-C mutant selective for VEGFR3) or physiological saline in controls, adenoviral therapy consisted of a single intraperitoneal injection on day 0 of adenoviral-5 vector (5×10 8 viral particles) encoding hVEGF-C C156S or lacZ as a control, and adeno-associated viral (AAV) gene delivery consisted of a single intraperitoneal injection 7 days before MI of AAV-9 vector (1×10 11 viral particles) encoding hVEGF-C C156S , sVEGFR3, or scrambled sequence as a control.

Techniques: Expressing, Imaging

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Lymphatic and Immune Cell Cross-Talk Regulates Cardiac Recovery After Experimental Myocardial Infarction

doi: 10.1161/ATVBAHA.120.314370

Figure Lengend Snippet: Adeno-associated viral (AAV)-sVEGFR3 (soluble vascular endothelial growth factor receptor 3) therapy inhibits infarct lymphangiogenesis postmyocardial infarction (MI). Mice were treated with AAV-hVEGF (human VEGF)-C C156S gene therapy (red circles, n=8), sVEGFR3 (AAV-sVEGFR3, blue triangles, n=5–7), or AAV-scrambled virus (MI controls, black circles, n=8), and sham-operated mice (white circle, n=7–9) served as healthy controls. Lymphangiogenesis in the infarct scar was evaluated as % proliferating lymphatic vessels ( A ), open lymphatic densities ( B ), and lymphatic area at 7 or 21 days post-MI ( C ). Examples ( D ) at 21 days post-MI of infarct zone left ventricular lymphatics (Lyve1 [lymphatic vessel endothelial receptor 1], red), macrophages (F4-80, gray), and cell nuclei (DAPI, blue); ×20 magnification, scale bar=50 µm. Note LYVE1-expressing macrophages close to lymphatic vessels. con indicates control.

Article Snippet: Modulation of cardiac lymphangiogenesis was performed using either systemic growth factor therapy with recombinant human VEGF-C C156S protein (RnD system) as described, or using viral gene vectors encoding hVEGF-C C156S (human VEGF-C C156S ) or sVEGFR3 (soluble VEGFR3)-IgG construct., , Briefly, protein therapy consisted of repeated (day 0, 2, 3, 4, and 6 post-MI) intraperitoneal injections of 2 µg/mouse (0.1 μg/g) of rhVEGF-C C156S (recombinant human VEGF-C mutant selective for VEGFR3) or physiological saline in controls, adenoviral therapy consisted of a single intraperitoneal injection on day 0 of adenoviral-5 vector (5×10 8 viral particles) encoding hVEGF-C C156S or lacZ as a control, and adeno-associated viral (AAV) gene delivery consisted of a single intraperitoneal injection 7 days before MI of AAV-9 vector (1×10 11 viral particles) encoding hVEGF-C C156S , sVEGFR3, or scrambled sequence as a control.

Techniques: Expressing

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Lymphatic and Immune Cell Cross-Talk Regulates Cardiac Recovery After Experimental Myocardial Infarction

doi: 10.1161/ATVBAHA.120.314370

Figure Lengend Snippet: Adeno-associated viral (AAV) delivery of VEGF (vascular endothelial growth factor)-CC156S reduces cardiac inflammation and improves cardiac function, while AAV-sVEGFR3 (soluble VEGF receptor 3) therapy reduces infarct scar T-cell levels and wall thinning leading to improved cardiac function in mice postmyocardial infarction (MI). Cardiac densities in the viable left ventricular (LV) of CD3 + T cells ( A ) and M1 pro-inflammatory macrophages (CD68 + /CD206 − cells), ( B ) were determined by immunohistochemistry in MI controls (AAV-scramble, black circles, n=8), and VEGFC (AAV–VEGF-C C156S , red circles, n=7–8) or sVEGFR3 (AAV-sVEGFR3, blue triangles, n=5–7) treated mice at 7 and 21 d post-MI. Healthy sham levels (n=7–9) are indicated by white circles / bars. Cardiac function and remodeling were analyzed by echocardiography at 21 days post-MI to determine LV fractional shortening ( C ) and LV anterior wall thickness in end-systole (AWT ES, D ). Infarct densities of CD3 + total T cells ( E ) and CD8 + T-cell subpopulation ( F ). Examples of CD8 + T-cell density in the infarct at 7 d post-MI ( G ). CD3 (red), CD8 (green), DAPI (blue), ×20 magnification, scale bar=50 µm. Infarct scar remodeling evaluated in Sirius red-stained histological sections as absolute infarct area ( H ) at 21 days post-MI in MI controls (AAV-scramble, n=8), VEGFC-(AAV-hVEGF-C C156S , n=7–8) and sVEGFR3-(AAV-sVEGFR3, n=5–7) treated mice. Examples of infarct scar remodeling. Scale bar=1 mm ( I ). LV hypertrophy/dilatation index was calculated from echocardiographic parameters at 21 days post-MI ( J ). Kruskal-Wallis, Dunn posthoc test. * P <0.05; ** P <0.01; *** P <0.001 vs sham; # P <0.05; ## P <0.01 vs MI control. con indicates control.

Article Snippet: Modulation of cardiac lymphangiogenesis was performed using either systemic growth factor therapy with recombinant human VEGF-C C156S protein (RnD system) as described, or using viral gene vectors encoding hVEGF-C C156S (human VEGF-C C156S ) or sVEGFR3 (soluble VEGFR3)-IgG construct., , Briefly, protein therapy consisted of repeated (day 0, 2, 3, 4, and 6 post-MI) intraperitoneal injections of 2 µg/mouse (0.1 μg/g) of rhVEGF-C C156S (recombinant human VEGF-C mutant selective for VEGFR3) or physiological saline in controls, adenoviral therapy consisted of a single intraperitoneal injection on day 0 of adenoviral-5 vector (5×10 8 viral particles) encoding hVEGF-C C156S or lacZ as a control, and adeno-associated viral (AAV) gene delivery consisted of a single intraperitoneal injection 7 days before MI of AAV-9 vector (1×10 11 viral particles) encoding hVEGF-C C156S , sVEGFR3, or scrambled sequence as a control.

Techniques: Immunohistochemistry, Staining

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Lymphatic and Immune Cell Cross-Talk Regulates Cardiac Recovery After Experimental Myocardial Infarction

doi: 10.1161/ATVBAHA.120.314370

Figure Lengend Snippet: Adeno-associated viral (AAV)-sVEGFR3 (soluble vascular endothelial growth factor receptor 3) therapy suppresses infarct lymphangiogenesis and improves cardiac function and remodeling in rats postmyocardial infarction (MI). Lymphatic remodeling was investigated in cardiac sections from MI controls (AAV-scramble, black circles, n=8) and sVEGFR3-treated (AAV-sVEGFR3, blue triangles, n=9) rats. Examples of blood vessels and lymphatics in the infarct zone at 28 d post-MI ( A ): middle panel: lymphatics (VEGFR3, red), and blood vessels (RECA1, green); bottom panel: lymphatics (Lyve1 [lymphatic vessel endothelial receptor 1], green; Prox1 [homeobox gene homologue Drosophilia prospero gene], red). ×20, scale bar=50 µm. White dashed lines outline infarct epicardium. Quantification of VEGFR3 + lymphatic area ( B ) and RECA1 + blood vascular area ( C ) in the infarct at 28 days post-MI. Comparison by Student 2-tailed t test: *** P <0.001 vs MI control. Cardiac function was investigated by serial echocardiography in healthy sham rats (open circles, n=4), MI controls (AAV-scramble, black circles, n=13), and sVEGFR3-treated (AAV-sVEGFR3, blue triangles, n=10) MI rats. Left ventricular (LV) fractional shortening ( D ), end-systolic diameter (ESD, E ), systolic interventricular septal wall thickness (IVSTs, F ) were measured, and LV hypertrophy/dilatation index ( G ) calculated. Two-way ANOVA, followed by Bonferroni posthoc: * P <0.05; ** P <0.01; *** P <0.001 vs sham; # P <0.05, ## P <0.01 vs MI control. RECA indicates rat endothelial cell antigen.

Article Snippet: Modulation of cardiac lymphangiogenesis was performed using either systemic growth factor therapy with recombinant human VEGF-C C156S protein (RnD system) as described, or using viral gene vectors encoding hVEGF-C C156S (human VEGF-C C156S ) or sVEGFR3 (soluble VEGFR3)-IgG construct., , Briefly, protein therapy consisted of repeated (day 0, 2, 3, 4, and 6 post-MI) intraperitoneal injections of 2 µg/mouse (0.1 μg/g) of rhVEGF-C C156S (recombinant human VEGF-C mutant selective for VEGFR3) or physiological saline in controls, adenoviral therapy consisted of a single intraperitoneal injection on day 0 of adenoviral-5 vector (5×10 8 viral particles) encoding hVEGF-C C156S or lacZ as a control, and adeno-associated viral (AAV) gene delivery consisted of a single intraperitoneal injection 7 days before MI of AAV-9 vector (1×10 11 viral particles) encoding hVEGF-C C156S , sVEGFR3, or scrambled sequence as a control.

Techniques: