Fig. 4

Aβ immunotherapy activated perivascular macrophages are enriched in inflammatory and extracellular matrix remodeling genes. (a) Schematic diagram showing macrophages activated by IgG2a or LALAPG immunodeficient control antibodies. (b) NanoString gene set enrichment analysis significance scores of macrophages activated IgG2a vs. control LALAPG. (c) Volcano plots showing directed differentially expressed matrix remodeling genes in IgG2a activated macrophages vs. control LALAPG activated macrophages. (d) NanoString generated heatmap of matrix remodeling gene expression (yellow = upregulation, purple = downregulation). (e) Volcano plots showing directed differentially expressed inflammatory signaling genes in IgG2a activated macrophages vs. control LALAPG activated macrophages. (f) NanoString generated heatmap of inflammatory signaling gene expression (yellow = upregulation, purple = downregulation). (g) Triple immunofluorescence of amyloid (Thio-S, green), Timp1 (red) and perivascular macrophages (CD169, cyan) in PDAPP mice treated with 3D6 or IgG control. Thio-S, Timp1 and CD169 immunoreactivity overlay (Merge). Colocalization analysis (white). (h) Quantification of Timp1⁺ area (%) of IgG or 3D6 treated mice. (i) Quantification of colocalization ratio of Timp1 and CD169. (j) Triple immunofluorescence of amyloid (Thio-S, green), MMP9 (red) and perivascular macrophages (CD169, cyan) in PDAPP mice treated with 3D6 or IgG control. Thio-S, MMP9 and CD169 immunoreactivity overlay (Merge). Colocalization analysis (white). (k) Quantification of MMP9⁺ area (%) of IgG or 3D6 treated mice. (l) Quantification of colocalization ratio of MMP9 and CD169. The number of vascular amyloid deposits analyzed was 8–10 per animal. Results are shown as ± SEM of n = 6 (mice). Asterisks indicate significant differences, where *** p < 0.001 and ****p < 0.0001 by unpaired Student’s t test. Horizontal line values correspond to adj. p-value < 0.01(solid), adj. p-value < 0.05(dashed). Scale bar 5 μm CC or 10 μm merge, respectively