anti saa1 2 antibody  (R&D Systems)

Journal: Stroke and vascular neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage.

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Figure 3 Human serum amyloid A1 (SAA1) protein levels in plasma and extracellular vesicles correlated (EVs) with clinical and laboratory assessments. (A, B) Bar graph indicating the concentration of SAA1 protein in both EVs and plasma for the ICH group and the healthy controls group. Unit: ng/mL in EVs and μg/mL in plasma. **p<0.01, ***p<0.001 by the two-tailed unpaired Student’s t-test and data are presented as means±SEM. (A) n=6, 15 in the control and ICH group, (B) n=13, 19 in the control and the ICH group. (C, D) Linear dependence graph revealing the relationship between neutrophils or leucocytes and the SAA1 protein in EVs (n=15). Correlation was analysed by spearman correlation analysis and the dashed line indicates the 95% CI. (E–H) Correlation analysis between the EVs-derived, plasma-derived SAA1 levels and the clinical assessments, including NIHSS and haemorrhage volume, through a linear dependence graph (n=15). Correlation was analysed by spearman correlation analysis and the dashed line indicates the 95% CI. ICH, intracerebral haemorrhage; NIHSS, National Institutes of Health Stroke Scale.

Article Snippet: We used anti- mouse SAA1 mAb (AF2948, NOVUS, USA) to block SAA1 in vivo at a dose of 0.25 μg/g, as previously reported.24 To establish a control, we administered Goat IgG Isotype Control (AB- 108- C, NOVUS, USA).

Techniques: Clinical Proteomics, Concentration Assay, Two Tailed Test, Control, Derivative Assay

Journal: Stroke and vascular neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage.

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Figure 4 Exogenous SAA1 upregulates the cell counts of microglia and astrocytes. (A) Schematic representation of exogenous SAA1 inserting into the basal ganglia region at varying dosages (750 ng, 250 ng, 83 ng and 0 ng). Images of immunostaining of microglia and astrocyte activation on day three are provided. (B, D) Images of immunostaining (B) and quantification (D) of microglia (stained by a specific marker, Iba1) around the ipsilateral basal ganglia region at different doses of exogenous SAA1. Microglia counts per mm2: 9.3±2.2, 41.5±3.9, 64.0±4.0, 88.3±8.8 for 0 ng, 83 ng, 250 ng, and 750 ng, respectively. (C, E) Images of immunostaining of GFAP (glial fibrillar acidic protein, known as the astrocyte active marker) representing astrocytes (C) and quantification of astrocytes (E) at different doses of exogenous SAA1 in the ipsilateral basal ganglia region. Astrocyte counts per mm2: 36.0±3.8, 93.5±6.1, 134.0±3.5, 176.5±11.6 for 0 ng, 83 ng, 250 ng, and 750 ng, respectively. Scale bars=100 µm, inset scale bars=20 µm, n=4 in each group. ***p<0.001 by the Kruskal-Wallis test. Data are presented as means±SEM.

Article Snippet: We used anti- mouse SAA1 mAb (AF2948, NOVUS, USA) to block SAA1 in vivo at a dose of 0.25 μg/g, as previously reported.24 To establish a control, we administered Goat IgG Isotype Control (AB- 108- C, NOVUS, USA).

Techniques: Immunostaining, Activation Assay, Staining, Marker

Journal: Stroke and vascular neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage.

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Figure 5 Blocking SAA1 promotes microglia reactivity and leucocyte infiltration. (A) Bar graph indicating the elevation of plasma SAA1 levels in ICH mice as compared with the sham group. n= 5 mice in the ICH group and n=10 mice in the sham group. **p<0.01 by two-tailed unpaired Student’s t-test. (B) Schematic diagram depicting intracerebral haemorrhage induction followed by intravenous administration of anti-SAA1 mAb or IgG 1 hour later. Immune cell populations were assessed using flow cytometry on days 1 and 3 post-ICH initiation. (C) Flow cytometry gating strategy depicting immune cell populations in mice brain treated with anti-SAA1 antibody or IgG on days 1 and day three post-ICH induction. The graph illustrates CD45high leucocytes, including CD3+CD19- T lymphocytes and its subtypes: CD4+CD8− T and CD4-CD8+ T lymphocytes, CD3−CD19+ B lymphocytes, CD11b+Ly6G+ neutrophils, and CD11b+F4/80+ macrophages. It also illustrates CD45int CD11b+microglia, including its subtypes: CD86+ microglia and CD206+ microglia. All gates were set using fluorescence-minus-one (FMO) controls. (D) Bar graph indicates the number of microglia and their subtypes in ICH mice with anti-SAA1 antibody or IgG treatment from days 1 to 3. (E) Bar graph shows major brain infiltrated leucocytes, involving CD8+ T lymphocytes, B lymphocytes, and neutrophils in ICH mice with anti-SAA1 antibody or IgG treatment from days 1 to 3. Int, intermediate. n=5, 6, 5 on day 1 and n=5, 13, 10 on day 3 for sham, IgG and mAb group. n=5, 9, 7 on day 3 for CD86+ microglia group in sham, IgG and mAb group specially. *p<0.05, **p<0.01, ***p<0.001 by one-way ANOVA and Tukey’s test. Data are presented as means±SEM. ANOVA, analysis of variance; ICH, intracerebral haemorrhage.

Article Snippet: We used anti- mouse SAA1 mAb (AF2948, NOVUS, USA) to block SAA1 in vivo at a dose of 0.25 μg/g, as previously reported.24 To establish a control, we administered Goat IgG Isotype Control (AB- 108- C, NOVUS, USA).

Techniques: Blocking Assay, Clinical Proteomics, Two Tailed Test, Flow Cytometry, Fluorescence

Journal: Stroke and vascular neurology

Article Title: Extracellular vesicles bearing serum amyloid A1 exacerbate neuroinflammation after intracerebral haemorrhage.

doi: 10.1136/svn-2024-003525

Figure Lengend Snippet: Figure 6 Anti-SAA1 mAb administration alleviates brain injury in mice with ICH. (A) Schematic diagram illustrating the ICH followed by intravenous injection of anti-SAA1 mAb or IgG 1 hour later. Subsequently, mice underwent MRI and neurological evaluations on days 1 and 3 after ICH induction. (B) Neurological scores of the sham group and the ICH group treated with anti- SAA1 mAb or IgG on day 1 and day 3. The modified Neurological Severity Score (mNSS) and rotarod test were used to measure the neurological deficit. n=6, 12, 7 for sham, IgG and mAb group. *p<0.05 by two-way ANOVA. (C) MRI of lesion volume (red) and perihaemorrhagic oedema (PHE) volume (yellow) on day 1 and day 3 post-ICH. (D, E) Quantification of lesion volume and PHE volume using the MRI. n=4, 6 for IgG and mAb group on day 1, n=5, 8 for IgG and mAb group on day 3. *p<0.05 by two-tailed unpaired Student’s t-test. Data are presented as means±SEM. ANOVA, analysis of variance; ICH, intracerebral haemorrhage.

Article Snippet: We used anti- mouse SAA1 mAb (AF2948, NOVUS, USA) to block SAA1 in vivo at a dose of 0.25 μg/g, as previously reported.24 To establish a control, we administered Goat IgG Isotype Control (AB- 108- C, NOVUS, USA).

Techniques: Injection, Modification, Two Tailed Test