Journal: The Journal of experimental medicine

Article Title: Chronic TREM2 activation exacerbates Aβ-associated tau seeding and spreading.

doi: 10.1084/jem.20220654

Figure Lengend Snippet: Figure 1. Chronic TREM2 activation increases DAM around plaques. (A) Schematic of experimental design. 6-mo-old 5XFAD mice were injected with AD- tau in the HC (bregma: −2.5 mm; lateral: −2.0 mm; depth: −2.2 mm) and overlying cortex (bregma: −2.5 mm; lateral: −2.0 mm; depth: −1.0 mm) and sacrificed 3 mo later to evaluate peri-plaque pathologies. 1 wk before AD-tau injection and every week following until sacrifice, 5XFAD mice were given i.p. injections of 80 mg/ml of the AL002a mouse TREM2 antibody (n = 13 female, n = 14 male) or the IgG control antibody (n = 14 female, n = 12 male). (B) Quantification of TREM2 antibody levels in terminal plasma for 5XFAD mice either chronically treated with the IgG control antibody or the TREM2 antibody. (C) Quantification of TREM2 antibody levels in plasma for 5XFAD mice either acutely treated with the IgG control antibody or the TREM2 antibody. (D) Quantification of TREM2 antibody levels in cortical tissue for 5XFAD mice either acutely treated with the IgG control antibody or the TREM2 antibody. For acute treatment, there were n = 8 IgG control and n = 9 Trem2 antibody–treated age- and sex-matched mice, unless otherwise specified on graphs. (E) Representative images of ipsilateral hemisphere stained with CLEC7A+ microglia, P2RY12+ microglia, and Aβ from IgG control and TREM2 antibody–treated groups. Scale bars, 15 µm. (F) Quantification of mouse sTREM2 levels in plasma for 5XFAD mice either acutely treated with the IgG control antibody or the TREM2 antibody. (G) Quantification

Article Snippet: Biotinylated rat anti-TREM2 antibody (R&D Systems) was added at 1:10,000 dilution in binding buffer and incubated for 1 h at RT.

Techniques: Activation Assay, Injection, Control, Clinical Proteomics, Staining

Journal: The Journal of experimental medicine

Article Title: Chronic TREM2 activation exacerbates Aβ-associated tau seeding and spreading.

doi: 10.1084/jem.20220654

Figure Lengend Snippet: Figure 2. Chronic TREM2 activation with a TREM2 antibody results in no changes in Aβ plaque burden. (A) Representative images of Aβ plaques in 5XFAD mice either treated with the IgG control antibody or the TREM2 antibody. Scale bar, 100 µm. (B–E) Quantification of Aβ staining in the ipsi- and contralateral cortices (B and D) and hippocampi (C and E) in female mice. (F–I) Quantification of Aβ staining in the ipsi- and contralateral cortices (F and H) and hippocampi (G and I) in male mice. Triangle symbol represents female mice, and square symbol represents male mice either treated with the IgG control antibody (n = 14 female, n = 12 male) or the TREM2 antibody (n = 13 female, n = 14 male). Data are presented as mean ± SEM. Scale bar, 100 µm. Significance was determined using a Student’s t test. ns, P > 0.05.

Article Snippet: Biotinylated rat anti-TREM2 antibody (R&D Systems) was added at 1:10,000 dilution in binding buffer and incubated for 1 h at RT.

Techniques: Activation Assay, Control, Staining

Journal: The Journal of experimental medicine

Article Title: Chronic TREM2 activation exacerbates Aβ-associated tau seeding and spreading.

doi: 10.1084/jem.20220654

Figure Lengend Snippet: Figure 3. Chronic TREM2 activation with a TREM2 antibody increases NP-tau pathology. (A) Representative images of ipsi- and contralateral hemi- spheres stained with AT8+ NP-tau pathology in AD-tau injected 5XFAD mice either treated with the IgG control antibody or the TREM2 antibody. Repre- sentative images are from female mice. Scale bars, 100 µm. (B–E) Quantification of p-tau (AT8+) staining in the ipsi- and contralateral cortices (B and D) and hippocampi (C and E) in female mice. (F–I) Quantification of p-tau (AT8+) staining in the ipsi- and contralateral cortices (F and H) and hippocampi (G and I) in male mice. Triangle symbol represents female mice, and square symbol represents male mice either treated with the IgG control antibody (n = 14 female, n = 12 male) or the TREM2 antibody (n = 13 female, n = 14 male). Data are presented as mean ± SEM. Significance was determined using a Student’s t test. ns, P > 0.05; *, P < 0.05.

Article Snippet: Biotinylated rat anti-TREM2 antibody (R&D Systems) was added at 1:10,000 dilution in binding buffer and incubated for 1 h at RT.

Techniques: Activation Assay, Staining, Injection, Control

Journal: The Journal of experimental medicine

Article Title: Chronic TREM2 activation exacerbates Aβ-associated tau seeding and spreading.

doi: 10.1084/jem.20220654

Figure Lengend Snippet: Figure 4. Chronic TREM2 activation with a TREM2 antibody increases peri-plaque NP-tau pathology and plaque-associated neuritic dystrophy, and acute TREM2 activation results in no changes in AD-tau uptake and degradation. (A) Representative images of BACE1+ and X34+ staining in ipsilateral HC. Scale bars, 20 µm. (B) Representative images of AT8+ and X34+ staining in ipsilateral HC. (C–F) Quantification of the number of BACE1+ staining within 15 µm of plaques in the ipsi- and contra- cortices (C and E) and hippocampi (D and F). (G–J) Quantification of the number of AT8+ staining within 15 µm of plaques in the ipsi- and contra- cortices (G and I) and hippocampi (H and J). (K) AD-tau uptake assay in TREM2 WT BMDMs. Results represent two independent ex- periments. (L) AD-tau degradation assay in TREM2 WT BMDMs. Results represent two independent experiments. Triangle symbol represents female mice, and square symbol represents male mice either treated with the IgG control antibody (n = 14 female, n = 12 male) or the TREM2 antibody (n = 13 female, n = 14

Article Snippet: Biotinylated rat anti-TREM2 antibody (R&D Systems) was added at 1:10,000 dilution in binding buffer and incubated for 1 h at RT.

Techniques: Activation Assay, Staining, Degradation Assay, Control

Journal: The Journal of experimental medicine

Article Title: Chronic TREM2 activation exacerbates Aβ-associated tau seeding and spreading.

doi: 10.1084/jem.20220654

Figure Lengend Snippet: Figure 5. Chronic TREM2 activation with a TREM2 antibody increases peri-plaque loss of synaptic marker synapsin. (A) Representative images of Synapsin+ and X34+ staining in ipsilateral HC. Scale bar, 7 µm. (B) Quantification of the number of Synapsin+ puncta within 15 µm of plaques in the ipsi-HC. (C–E) Quantification of the number of Synapsin+ puncta within 15 µm of plaques in the ipsi-cortex, contra-cortex, and HC. Triangle symbol represents female mice, and square symbol represents male mice either treated with the IgG control antibody (n = 14 female, n = 12 male) or the TREM2 antibody (n = 13 female, n = 14 male). Data are presented as mean ± SEM. Significance was determined using a linear regression with sex as a covariate. ns, P > 0.05; *, P < 0.05.

Article Snippet: Biotinylated rat anti-TREM2 antibody (R&D Systems) was added at 1:10,000 dilution in binding buffer and incubated for 1 h at RT.

Techniques: Activation Assay, Marker, Staining, Control