Anti-Human Abeta Recombinant Antibody (Ponezumab) (CAT#: TAB-227)

Figure 1 Quantification of individual cerebral vessels intravital multiphoton microscopy.

Quantification of cerebral vessels from PSAPP mice (6 months of age,n= 6–7 per group) imaged over a 3-month period and following chronic administration of ponezumab (blue, 10 mg/kg weekly, intraperitoneal) or vehicle (red). For the normalized fraction of positive voxels we found that only time was a significant covariate. Planned comparisons revealed that relative to baseline, the level was significantly higher at 1 month [t(56) = 3.45,P50.003] and at 3 months for the vehicle treated group only. Following ponezumab administration there was no statistically significant differences [t(56) = 1.5,P50.29 andt(56) = 1.04, P<0.55, for 1 and 3 months, respectively]. Data are represented as mean SEM. ***P<0.001 (A). Representative images following dextran labelling of vasculature (blue) with methoxy-034-labelled amyloid (magenta) following vehicle (B) or ponezumab administration (C) for 3 months. Scale bar = 100mm.

Bales, K. R., O’neill, S. M., Pozdnyakov, N., Pan, F., Caouette, D., Pi, Y., ... & Johnson, A. W. (2015). Passive immunotherapy targeting amyloid-β reduces cerebral amyloid angiopathy and improves vascular reactivity. Brain, 139(2), 563-577.

Figure 2 Quantification of amyloid-band amyloid in cerebral vessels following administration of ponezumab.

Significant reduction in Congo red-positive amyloid in leptomeningeal (A) and brain vessels (B). Significant reduction in the number of leptomeningeal (C)or brain vessels (D) immunoreactive for amyloid-b40following administration of ponezumab. PSAPP mice (6 months of age at study start) were treated for 6 months with ponezumab (10 mg/kg; weekly intraperitoneal; n= 6–9 mice per group). The number of vessels (average number/section/mouse) that were stained (550% coverage) were plotted for each section analysed (6–9 sections analysed per animal). Data are represented as mean SEM. *P<0.05, **P<0.01. Representative lower power image of mid (E) and rostral (F) sections stained with antiamyloid-b40antibody (Scale bar = 500mm) from vehicle treated group. High power surface (e) and parenchymal (f) vessel (Scale bar 25 =mm).

Bales, K. R., O’neill, S. M., Pozdnyakov, N., Pan, F., Caouette, D., Pi, Y., ... & Johnson, A. W. (2015). Passive immunotherapy targeting amyloid-β reduces cerebral amyloid angiopathy and improves vascular reactivity. Brain, 139(2), 563-577.

Figure 3 Enrichment of cerebral vasculature elements from brain tissue and quantification of amyloid-bpeptides following administration of ponezumab.

Enrichment of cerebral vascular elements from brain tissue (A, BH = crude brain homogenate, S = neuronal elements; V = cerebrovascular enriched fraction; eNOS = endothelial nitric oxide, encoded byNos3; PECAM = platelet/endothelial cell adhesion molecule 1; GAPDH = glyceraldehyde dehydrogenase). Significant reduction in detergent soluble amyloid-b40 (B) and amyloid-b42 (C) following 3 months treatment of PSAPP mice with ponezumab (6 months of age at study start; 10 mg/kg, weekly intraperitoneal), *P<0.05. There was no significant reduction in amyloid-b40 (D) or amyloid-b42 (E) levels from the guanidine extractable pool measured from the cerebrovascular enriched fraction;n= 6–7 per group.

Bales, K. R., O’neill, S. M., Pozdnyakov, N., Pan, F., Caouette, D., Pi, Y., ... & Johnson, A. W. (2015). Passive immunotherapy targeting amyloid-β reduces cerebral amyloid angiopathy and improves vascular reactivity. Brain, 139(2), 563-577.

Figure 4 Interstitial fluid levels of amyloid-b40.

Following infusion of a potent gamma secretase inhibitor (Cmpd E; 200 nM; GSI) and peripheral administration of ponezumab (10 mg/kg; intraperitoneal) to APP transgenic mice there was a significant increase in interstitial fluid amyloid-blevels in old plaque bearing mice only (A). Quantification of the change from basal interstitial fluid (ISF) levels over a 30.5–44-h time period after gamma secretase inhibitor infusion and ponezumab administration (B). There is a significant correlation between basal interstitial fluid amyloid-b40 levels and total hippocampal plaque load (both parenchymal and vascular) proximal to the microdialysis probe following treatment (C). (APP = APP K670N;M671L old/young = 18/6 months of age; APP/PS1 = APP K670N;M671L/PS1deltaE9; 12 months of age), n= 3–6 per group (Cirritoet al., 2003).

Bales, K. R., O’neill, S. M., Pozdnyakov, N., Pan, F., Caouette, D., Pi, Y., ... & Johnson, A. W. (2015). Passive immunotherapy targeting amyloid-β reduces cerebral amyloid angiopathy and improves vascular reactivity. Brain, 139(2), 563-577.