mouse anti 267 trem2 2b5  (R&D Systems)

Journal: Journal of Alzheimer's Disease

Article Title: Choroid Plexus Acts as Gatekeeper for TREM2, Abnormal Accumulation of ApoE, and Fibrillary Tau in Alzheimer’s Disease and in Down Syndrome Dementia

doi: 10.3233/jad-181179

Figure Lengend Snippet: Fig. 1. Plasma ApoE, soluble TREM, and A42 protein levels were declined in DS with dementia progression. Plasma protein levels of TREM2, ApoE, tau, A42, and A40 in DS and age-matched controls were determined by ELISA. Levels of sTREM2 were found to be significantly higher in controls compared to DS; R2 = 0.78, p ≤0.0001 (a). Soluble TREM2 level is age dependent, decreasing with age and disease progression in DS. Horizontal scattered bars indicate median sTREM2 concentration per group (b). The levels of ApoE was widely distributed in DS, R2 = 0.44, p ≤0.001 (c). ApoE plasma levels were higher in older controls compared to age-matched older DS as shown by scattered plot (d). ApoE 2 carriers have higher ApoE levels than ApoE 4 heterozygous subjects, R2 = 0.78, p ≤0.0001 (e). The total tau (t-Tau) level was not significantly different between Y-DS compared to O-DS, R2 = 0.38, p ≤0.01 (f) and shown by scattered plot (h, i). Whereas p-tau was significantly higher in O-DS subjects compared to Y-DS, R2 = 0.45, p ≤0.001 (g), and as shown by scattered plot (j). The plasma A40 level was found to be increased with age, R2 = 0.77, p ≤0.0001 (k), and A42 level was decreased with age, R2 = 0.56, p ≤0.0001 (l). The A42/A40 ratio, R2 = 0.66, p ≤0.0001, decreased with age and dementia progression in DS (m).

Article Snippet: The specificity of the used ELISA262 system used was further validated by anti-TREM2263 immunoblotting showing a high degree of correlation264 between the ELISA readings and immunoreactiv-265 ity on the immunoblot using these antibodies and266 an independent anti-TREM2 antibody, mouse anti-267 TREM2 2B5 (R&D Systems, Minneapolis, MN).268 The plasma was stored at –70◦C until used for269 further assays.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Biomarker Discovery, Concentration Assay

Journal: Journal of Alzheimer's Disease

Article Title: Choroid Plexus Acts as Gatekeeper for TREM2, Abnormal Accumulation of ApoE, and Fibrillary Tau in Alzheimer’s Disease and in Down Syndrome Dementia

doi: 10.3233/jad-181179

Figure Lengend Snippet: Fig. 2. TREM2 and tau proteins did not co-localize with A42 senile plaques in mid temporal cortex. Double immunofluorescence staining and confocal images were performed on the mid temporal cortex (MTC) of AD and DS brain sections using rabbit monoclonal anti-A42 (rab-mAb A42, green) and mAb anti-phospho-tau (AT8, red) antibody, DAPI for nuclear staining (Blue). A42 immunoreactivity was visible in the senile plaques (SPs) but did not co-localized with p-Tau (a–e). In DS brain (DS8), the layer III of MTC stained for anti-A42 (green) showed many mature plaques (f and h). TREM2 was only present around the SPs and visible in the neuropil thread (f and g). Further staining of different cases of AD (AD3 and AD5, Supplementary Table 1) and images was captured by confocal microscopy, showed that mature A42 positive SPs did not co-localize with p-Tau positive neurofibrillary tangles. Scale bar in a–c = 50 m, d-k = 25 m, and l = 10 m.

Article Snippet: The specificity of the used ELISA262 system used was further validated by anti-TREM2263 immunoblotting showing a high degree of correlation264 between the ELISA readings and immunoreactiv-265 ity on the immunoblot using these antibodies and266 an independent anti-TREM2 antibody, mouse anti-267 TREM2 2B5 (R&D Systems, Minneapolis, MN).268 The plasma was stored at –70◦C until used for269 further assays.

Techniques: Staining, Confocal Microscopy

Journal: Journal of Alzheimer's Disease

Article Title: Choroid Plexus Acts as Gatekeeper for TREM2, Abnormal Accumulation of ApoE, and Fibrillary Tau in Alzheimer’s Disease and in Down Syndrome Dementia

doi: 10.3233/jad-181179

Figure Lengend Snippet: Fig. 3. Increased insoluble tau in the hippocampus and in entorhinal cortex of DS brains. Confocal microscopy analysis was performed on controls, AD, and DS brain sections particularly in the hippocampus (HP) and entorhinal cortex. Total anti-Tau (HT7) for control brains and anti-p-Tau (AT8) for AD and DS were used and DAPI for nuclear staining. In a young control brain (C1, Supplementary Table 1), t-Tau (recognized by HT7, green) and TREM2 (red) co-localized in the pyramidal neurons of HP, in dentate gyrus granule cells, subiculum, cortical, and subcortical areas (a–d). In a younger DS subject (DS3-b, Braak stage 2, Supplementary Table 1), TREM2 was visible in the dentate gyrus granule cells and insoluble p-Tau in the pyramidal neurons (in the axons) with limited co-localization (e-g). In AD brains, limited TREM2 expression was seen in the HP and did not co-localized with Iba1 positive activated microglia (f). Confocal images of a DS brain (DS3), TREM2, and p-Tau co-localized in the axons (g-i), but TREM2 was not visible in neurofibrillary tangles (j). In an AD brain section (AD2), TREM2 proteins were present in the cell body, whereas p-Tau was visible in the neuritic threads, neurofibrillary tangles, and damaged axons (k). TREM2 levels were highest in controls and lowest in AD brains (l, p ≤0.0001). Scale bar in a–f = 50 m, g–k = 25 m. Image intensity measured by Image J. Error bars indicate SEM, ∗∗p < 0.001, ∗∗∗p < 0.0001.

Article Snippet: The specificity of the used ELISA262 system used was further validated by anti-TREM2263 immunoblotting showing a high degree of correlation264 between the ELISA readings and immunoreactiv-265 ity on the immunoblot using these antibodies and266 an independent anti-TREM2 antibody, mouse anti-267 TREM2 2B5 (R&D Systems, Minneapolis, MN).268 The plasma was stored at –70◦C until used for269 further assays.

Techniques: Confocal Microscopy, Control, Staining, Expressing

Journal: Journal of Alzheimer's Disease

Article Title: Choroid Plexus Acts as Gatekeeper for TREM2, Abnormal Accumulation of ApoE, and Fibrillary Tau in Alzheimer’s Disease and in Down Syndrome Dementia

doi: 10.3233/jad-181179

Figure Lengend Snippet: Fig. 4. TREM2 expression was reduced in white matter tracts in DS brain. White matter tracts (WMTs) are narrowed in DS brains as shown by PET imaging data. Thus, WMTs and corpus callosum (CC) of control, AD, and DS brain sections were analyzed by immunofluorescence using TREM2, t-Tau, or p-Tau antibodies. In control brains in the superior frontal cortex (grey matter), TREM2 (red) and t-Tau (HT7, green) co-localized in cortical neurons (a–c), whereas in striatum (STR), WM-tract both proteins were present in oligodendrocytes (d). In DS and AD brains, p-Tau (stained with AT8) were present in the in the neuropils and in neurofibrillary tangles and did not co-localize with TREM2 (e and f). In control brains, t-Tau protein was present in the WMT of STR and in the CC (g). In DS brain in STR, WMT was damaged and high infiltration of Iba1 positive microglial cells were visible (h). TREM2 did not co-localize with p-Tau positive SP in AD brain sections (i). Confocal image of DS brain, WMTs of the CC were shrunk (atrophic), and some MBP positive oligodendrocytes were co-localized with TREM2 (j-l). Both proteins (TREM2 and p-Tau) were detected in WMT in DS and AD cases but with reduced expression in DS (m-n). TREM2 levels were higher in WMT of control and AD compared to DS (o, p ≤0.001). Scale bar in a–c = 75 m, d–i, m, n = 25 m, j–l = 20 m. Image intensity measured by Image J. Error bars indicate SEM, ∗∗p < 0.001.

Article Snippet: The specificity of the used ELISA262 system used was further validated by anti-TREM2263 immunoblotting showing a high degree of correlation264 between the ELISA readings and immunoreactiv-265 ity on the immunoblot using these antibodies and266 an independent anti-TREM2 antibody, mouse anti-267 TREM2 2B5 (R&D Systems, Minneapolis, MN).268 The plasma was stored at –70◦C until used for269 further assays.

Techniques: Expressing, Imaging, Control, Staining

Journal: Journal of Alzheimer's Disease

Article Title: Choroid Plexus Acts as Gatekeeper for TREM2, Abnormal Accumulation of ApoE, and Fibrillary Tau in Alzheimer’s Disease and in Down Syndrome Dementia

doi: 10.3233/jad-181179

Figure Lengend Snippet: Fig. 5. Phospho-tau aggregates and ApoE were visible in the fenestrated capillaries of choroid plexus and sTREM2 in the macrophages. Brain sections containing choroid plexus (CP) in the lateral ventricles and third ventricles from control, AD, and DS subjects were stained with anti-TREM2 and anti-Tau (t-Tau or p-Tau), or anti-TREM2 and anti-ApoE antibodies and analyzed by confocal microscopy. DAPI was used for nuclear staining. In control brain, both TREM2 and anti-t-Tau were present in CP cuboidal epithelial cells (CPE) surrounding a core of fenestrated capillaries and connective tissues (a). Some soluble t-Tau protein was visible inside fenestrated capillaries and TREM2 was visible in stromal capillary (vesicles) and in the stromal macrophages (a and b). In DS and AD brain dense fibrillary p-Tau present in psammoma bodies (calcified intracellular inclusion structures), was visible in the stroma and TREM2 co-localized in the vesicles and in stromal macrophages (c and d). In control CP, both proteins TREM2 (red) and ApoE (green) were visible in CPE cells and in stromal macrophages that appeared normal with healthy structure (e). In contrast, in DS brains ApoE was visible in the “amyloid biondi” bodies (complex filamentous ring-like structures associated with lipid droplets, showing with an arrow) and lipofuscin (yellow or brown intracellular structures composed of lipid molecules (f). Scale bar in a = 50 m, b–f = 25 m.

Article Snippet: The specificity of the used ELISA262 system used was further validated by anti-TREM2263 immunoblotting showing a high degree of correlation264 between the ELISA readings and immunoreactiv-265 ity on the immunoblot using these antibodies and266 an independent anti-TREM2 antibody, mouse anti-267 TREM2 2B5 (R&D Systems, Minneapolis, MN).268 The plasma was stored at –70◦C until used for269 further assays.

Techniques: Control, Staining, Confocal Microscopy