Human Anti-APP Recombinant Antibody (clone hu3D6) (CAT#: HPAB-S0004-YC)

Figure 1 Topical antibody application leads to clearance of diffuse amyloid-β in Tg2576 mice.

These images are projections of threedimensional volumes from the cortex of a representative Tg2576 mouse treated with FITC-labeled 10d5 antibody. The images were acquired in the anesthetized mouse using multiphoton microscopy. The left shows labeled amyloid deposits at the initial application of antibody. Numerous diffuse deposits as well as amyloid angiopathy can be seen. The right is the same volume 3 d later, labeled with 3d6 antibody, which recognizes amyloid-β independently of 10d5. A majority of the amyloid-β deposits have been cleared in this 3 d period.

Bacskai, B. J., Kajdasz, S. T., McLellan, M. E., Games, D., Seubert, P., Schenk, D., & Hyman, B. T. (2002). Non-Fc-mediated mechanisms are involved in clearance of amyloid-β in vivo by immunotherapy. Journal of Neuroscience, 22(18), 7873-7878.

Figure 2 10d5 and 3d6 antibodies colocalize at noncompetitive binding sites on amyloid-β plaques.

Paraformaldehyde-fixed tissue sections from an 18-month-old Tg2576 mouse brain were treated sequentially with FITC-labeled 10d5 antibodies, followed by rhodamine-labeled 3d6 antibodies. Images were obtained with a Bio-Rad confocal microscope using 488 nm excitation for FITC (A) and 568 nm excitation for rhodamine (B). A color-merged image (C) shows that 10d5 (green) and 3d6 (red) are both able to bind to the plaques and colocalize everywhere, producing a yellow color. Scale bar, 50 μm.

Bacskai, B. J., Kajdasz, S. T., McLellan, M. E., Games, D., Seubert, P., Schenk, D., & Hyman, B. T. (2002). Non-Fc-mediated mechanisms are involved in clearance of amyloid-β in vivo by immunotherapy. Journal of Neuroscience, 22(18), 7873-7878.

Figure 3 10d5 and 3d6 antibodies are equally effective at clearing diffuse amyloid-β deposits in the PDAPP mouse model after 3 d.

10d5 and 3d6 antibodies was similarly effective after 3 d when applied topically to the PDAPP mouse.

Bacskai, B. J., Kajdasz, S. T., McLellan, M. E., Games, D., Seubert, P., Schenk, D., & Hyman, B. T. (2002). Non-Fc-mediated mechanisms are involved in clearance of amyloid-β in vivo by immunotherapy. Journal of Neuroscience, 22(18), 7873-7878.

Figure 4 Remaining dense-core plaques in PDAPP mice are reduced in size 3 d after treatment with anti-amyloid-β antibodies.

The identified plaques that were imaged in the first imaging session and remained after 3 d after a single antibody treatment were measured as described previously. Plaques that were cleared completely would have been measured as 100% change in size but are not included in this analysis. 10d5, 3d6, and F(ab')2 fragments of 3d6 were equally effective at reducing the size of remaining dense-core plaques compared with control antibody 16b5.

Bacskai, B. J., Kajdasz, S. T., McLellan, M. E., Games, D., Seubert, P., Schenk, D., & Hyman, B. T. (2002). Non-Fc-mediated mechanisms are involved in clearance of amyloid-β in vivo by immunotherapy. Journal of Neuroscience, 22(18), 7873-7878.

Figure 5 The 3D6 treatment does not improve cognition in APP/PS1 mice on the HHcy diet.

Two day radial arm water maze data are shown for WT mice (A) and APP/PS1 mice (B). The mean number of errors per trial was calculated for each block; each block is the average of three trials. Block 10 data are graphed for the WT mice (C) and the APP/PS1 mice (D).

Weekman, E. M., Sudduth, T. L., Caverly, C. N., Kopper, T. J., Phillips, O. W., Powell, D. K., & Wilcock, D. M. (2016). Reduced efficacy of anti-Aβ immunotherapy in a mouse model of amyloid deposition and vascular cognitive impairment comorbidity. Journal of Neuroscience, 36(38), 9896-9907.

Figure 6 Total Aβ is reduced by 3D6 treatment.

Representative images of A staining in the hippocampus of APP/PS1, control, and IgG2a (A), APP/PS1, control, and 3D6 (B), APP/PS1, HHcy, and IgG2a (C), and APP/PS1, HHcy, and 3D6 (D). A, The cornu ammonis (CA) 1, CA3, and dentate gyrus (DG) are labeled for orientation. Scale bar: A, 120μm. E, Quantification of percentage positivestain in the frontal cortex and hippocampus. F, Biochemical quantification of soluble and insoluble Aβ1-38, Aβ1-40.

Weekman, E. M., Sudduth, T. L., Caverly, C. N., Kopper, T. J., Phillips, O. W., Powell, D. K., & Wilcock, D. M. (2016). Reduced efficacy of anti-Aβ immunotherapy in a mouse model of amyloid deposition and vascular cognitive impairment comorbidity. Journal of Neuroscience, 36(38), 9896-9907.

Figure 7 HHcy redistributes amyloid to the vasculature.

Representative images of Congo red staining in the hippocampus of APP/PS1, control, and IgG2a (A), APP/PS1, control, and 3D6(B), APP/PS1, HHcy, and IgG2a (C), and APP/PS1, HHcy, and 3D6 (D). A, The CA1, CA3, and DG are labeled for orientation. Representative images of Congo red staining in the cortex of APP/PS1, control, and IgG2a (E), APP/PS1, control, and 3D6 (F), APP/PS1, HHcy, and IgG2a (G), and APP/PS1, HHcy, and 3D6 (H).

Weekman, E. M., Sudduth, T. L., Caverly, C. N., Kopper, T. J., Phillips, O. W., Powell, D. K., & Wilcock, D. M. (2016). Reduced efficacy of anti-Aβ immunotherapy in a mouse model of amyloid deposition and vascular cognitive impairment comorbidity. Journal of Neuroscience, 36(38), 9896-9907.

Figure 8 The 3D6 increases MRI detected microhemorrhages.

Microhemorrhage numbers were significantly greater in the APP/PS1 mice on the HHcy or control diet receiving 3D6 antibody than either the APP/PS1 mice on control diet or the HHcy diet receiving IgG2a treatment.

Weekman, E. M., Sudduth, T. L., Caverly, C. N., Kopper, T. J., Phillips, O. W., Powell, D. K., & Wilcock, D. M. (2016). Reduced efficacy of anti-Aβ immunotherapy in a mouse model of amyloid deposition and vascular cognitive impairment comorbidity. Journal of Neuroscience, 36(38), 9896-9907.

Figure 9 The 3D6 and HHcy increase Prussain blue detected microhemorrhages.

A, Representative images of Prussian blue-positive microhemorrhages in the frontal cortex. Scale bar, 120 μm. B, Quantification of the mean number of microhemorrhages per section. **p <0.01, compared with APP/PS1, control, and IgG2a. Black bars represent significant differences between connecting groups. Prussian blue histological analysis of microhemorrhages showed a significant increase in the number of microhemorrhages in each of the groups compared with APP/PS1 mice on control diet with IgG2a treatment.

Weekman, E. M., Sudduth, T. L., Caverly, C. N., Kopper, T. J., Phillips, O. W., Powell, D. K., & Wilcock, D. M. (2016). Reduced efficacy of anti-Aβ immunotherapy in a mouse model of amyloid deposition and vascular cognitive impairment comorbidity. Journal of Neuroscience, 36(38), 9896-9907.

Figure 10 CD11b staining is decreased in the HHcy groups.

Representative images of CD11b staining in the hippocampus of APP/PS1, control, and IgG2a (A), APP/PS1, control, and 3D6 (B), APP/PS1, HHcy, and IgG2a (C), and APP/PS1, HHcy, and 3D6 (D). A, The CA1, CA3, and DG are labeled for orientation. Scale bar: A, 120 μm.

Weekman, E. M., Sudduth, T. L., Caverly, C. N., Kopper, T. J., Phillips, O. W., Powell, D. K., & Wilcock, D. M. (2016). Reduced efficacy of anti-Aβ immunotherapy in a mouse model of amyloid deposition and vascular cognitive impairment comorbidity. Journal of Neuroscience, 36(38), 9896-9907.

Figure 11 HHcy reduced inflammatory markers and increased the MMP system markers in both the IgG2a and 3D6 groups.

Data are shown as a fold change from APP/PS1, control, and IgG2a. Relative gene expression for proinflammatory markers (A), would healing/repair markers (B), immune complex-mediated markers (C), and MMP2 and MMP9 system markers (D). *p<0.05, compared with APP/PS1, control, and IgG2a. **p<0.01, compared with APP/PS1, control, and IgG2a.

Weekman, E. M., Sudduth, T. L., Caverly, C. N., Kopper, T. J., Phillips, O. W., Powell, D. K., & Wilcock, D. M. (2016). Reduced efficacy of anti-Aβ immunotherapy in a mouse model of amyloid deposition and vascular cognitive impairment comorbidity. Journal of Neuroscience, 36(38), 9896-9907.