clone 22d1  (Bio X Cell)

Journal: Virology

Article Title: Prion pathogenesis is unaltered following down-regulation of SIGN-R1

doi: 10.1016/j.virol.2016.08.005

Figure Lengend Snippet: Transient down-regulation of SIGN-R1 on MZ macrophages. Panel A. Mice were injected IV with anti-SIGN-R1-specific mAb 22D1 or control Ig and spleens collected 24 h later. In spleens from control Ig-treated mice (upper row) MZ macrophages expressing SIGN-R1 (green, detected with anti- SIGN-R1 mAb clone ER-TR9) and MARCO (red) were readily detected in the MZ. SIGN-R1 expression on MZ macrophages was down-regulated in the spleens of anti-SIGN-R1 mAb-treated mice (lower row). Anti-SIGN-R1 mAb treatment did not affect the expression of CD169 on the marginal metallophilic macrophages (blue, right-hand panels). Panel B. Twenty four hours after antibody treatment mice were injected IV with FITC-70 kDa dextran (FITC-dextran, green) and spleens collected 24 h later. MZ macrophages (MARCO + cells, red) in the spleens of anti-SIGN-R1 mAb-treated mice were unable to retain FITC-dextran (lower row). Dotted lines indicate the boundary of the MZ. n =6 mice/group.

Article Snippet: To transiently down-regulate SIGN-R1 expression in vivo , mice were injected IV with 100 μg of hamster anti-mouse SIGN-R1 monoclonal antibody (mAb; clone 22D1, eBioscience, Hatfield, UK) as described ( , , ).

Techniques: Injection, Control, Expressing

Journal: Virology

Article Title: Prion pathogenesis is unaltered following down-regulation of SIGN-R1

doi: 10.1016/j.virol.2016.08.005

Figure Lengend Snippet: Transient down-regulation of SIGN-R1 on MZ macrophages does not adversely affect FDC status. Panel A. IHC analysis suggested there was no observable difference in the expression of CR1/CD35 (red) or cellular PrP C (blue) in FDC in spleens from anti-SIGN-R1 mAb- or control Ig-treated mice. Panels B and C. Morphometric analysis confirmed that the magnitude of the CR1/CD35- and PrP C -specific immunostaining observed in the spleens from the anti-SIGN-R1 mAb or control Ig mice was similar. Panel D. Mice were injected IV with anti-SIGN-R1-specific mAb 22D1 or control Ig, and 24 h later passively immunized with preformed PAP immune complexes ( n =6 mice/group). Twenty four hours after treatment the presence of immune complexes (PAP, green) upon FDC (CR1/CD25 + cells, red) was determined by IHC. Panel E. Morphometric analysis suggested the amount of PAP trapped on the surfaces of the FDC in the spleens of anti-SIGN-R1 mAb-treated mice was significantly greater when compared to control Ig-treated mice ( P <0.0001).

Article Snippet: To transiently down-regulate SIGN-R1 expression in vivo , mice were injected IV with 100 μg of hamster anti-mouse SIGN-R1 monoclonal antibody (mAb; clone 22D1, eBioscience, Hatfield, UK) as described ( , , ).

Techniques: Expressing, Control, Immunostaining, Injection

Journal: Virology

Article Title: Prion pathogenesis is unaltered following down-regulation of SIGN-R1

doi: 10.1016/j.virol.2016.08.005

Figure Lengend Snippet: Transient down-regulation of SIGN-R1 on MZ macrophages on does not impair the accumulation of PrP Sc upon FDC in the spleen. Mice were injected IV with anti-SIGN-R1- mAb or control Ig and 24 h later injected IV with ME7 scrapie prions. Spleens were collected 35 days after IV prion injection (Panel A) or at the terminal stage of disease (Panel B). At each time point abundant prion disease-specific PrP (PrP d , brown, left-hand column) accumulated in association with FDC (CD21/35 + cells, brown, middle column) in the spleens of mice from each treatment group. Analysis of adjacent sections by PET immunoblot analysis confirmed the presence of prion-disease specific, relatively proteinase K-resistant PrP Sc (black, right-hand column).

Article Snippet: To transiently down-regulate SIGN-R1 expression in vivo , mice were injected IV with 100 μg of hamster anti-mouse SIGN-R1 monoclonal antibody (mAb; clone 22D1, eBioscience, Hatfield, UK) as described ( , , ).

Techniques: Injection, Control, Western Blot

Journal: Virology

Article Title: Prion pathogenesis is unaltered following down-regulation of SIGN-R1

doi: 10.1016/j.virol.2016.08.005

Figure Lengend Snippet: Transient down-regulation of SIGN-R1 on MZ macrophages at the time of IV prion infection does affect the development of neuropathology in the brain at the terminal stage of disease. Mice were injected IV with anti-SIGN-R1 mAb or control Ig and 24 h later injected IV with ME7 scrapie prions ( n =8 mice/group). Brains were collected at the terminal stage of disease. Panel A. Histopathological analysis showed large accumulations of prion disease-specific PrP d (brown, upper row), reactive astrocytes expressing GFAP (brown, second row), active microglia expressing AIF1/Iba1 (brown, third row) and spongiform pathology (H&E, bottom row) in brains of all terminally-affected control Ig-treated (left-hand column) and anti-SIGN-R1 mAb-treated (right-hand column) mice. Sections were counterstained with haematoxylin to detect cell nuclei (blue). Panel B. Immunoblot analysis of brain tissue homogenates confirmed the presence of high levels of prion-specific, relatively proteinase K (PK)-resistant PrP Sc within the brains of mice from each treatment group. Samples were treated in the presence (+) or absence (−) of PK before electrophoresis. After PK treatment, a typical three-band pattern was observed between molecular mass values of 20–30 kDa, representing unglycosylated, monoglycosylated, and diglycosylated isomers of PrP (in order of increasing molecular mass). Panel C. The severity and distribution of the spongiform pathology (vacuolation) within the brains of all terminally-affected mice from each treatment group was similar. The severity of the vacuolation in each brain was scored on a scale of 1–5 in the following grey matter regions: G1, dorsal medulla; G2, cerebellar cortex; G3, superior colliculus; G4, hypothalamus; G5, thalamus; G6, hippocampus; G7, septum; G8, retrosplenial and adjacent motor cortex; G9, cingulate and adjacent motor cortex.

Article Snippet: To transiently down-regulate SIGN-R1 expression in vivo , mice were injected IV with 100 μg of hamster anti-mouse SIGN-R1 monoclonal antibody (mAb; clone 22D1, eBioscience, Hatfield, UK) as described ( , , ).

Techniques: Infection, Injection, Control, Expressing, Western Blot, Electrophoresis