Table 2 Markers of Inflammation and Immunoreactivity in Cerebral Arteries and Arterioles in CADASIL
From: ER stress induced immunopathology involving complement in CADASIL: implications for therapeutics
Marker (antibody, source) | Description and Source | Change in marker | CADASIL | Controls | Disease Controls†(SVD) |
|---|---|---|---|---|---|
(0, +(1),-++(2), +++(3) score of immunoreactivities or % (SD) of vessels positive for marker) | |||||
CADASIL Arteriopathy | |||||
SMA | α-smooth muscle actin (monoclonal, 1:3,000; Sigma) | ↓Cytoskeletal component | +++ | + | +++ |
Fractin | Actin filament fragment (polyclonal, 1:100, gift from Dr | ↑ECM | +++ | + | +++ |
Medin | Smooth muscle protein (polyclonal, 1:100, gift from Dr | ↑ECM | +++ | + | +++ |
GLUT1 | Glucose Transporter 1 (polyclonal, 1:1,000; Thermo Scientific) | ↓Endothelial cells | ++ | + | ++ |
COL1 | Collagen I (polyclonal,1:1,000; Dako) | ↑ECM | +++ | + | ++ |
COL3 | Collagen III (polyclonal, 1:1,000; Dako) | ↑ECM | +++ | + | ++ |
COL4 | Collagen IV (polyclonal, 1:1,000, Sigma) | ↑ECM | +++ | + | ++ |
Fibrinogen | Fibrinogen (polyclonal, 1:2,000; Dako) | ↑Plasma protein | ++ | + | +++ |
Perivascular microglia/macrophages | |||||
CD45 | CD45 (monoclonal, 1:1,000; Dako) | ↑Microglial/macrophage cells | +++ | + | + |
CD68 | CD68 (monoclonal, 1:400, Clone PG-M1, DAKO) | ↑Microglial/macrophage cells | +++ | ++ | ++ |
CD163 | CD163 (monoclonal, 1:1,000; Dako) | ↑Macrophage cells | +++ | - | + |
Iba-1 | Iba-1 (polyclonal, 1:1,000, 66827-1-Ig, Proteintech) | Microglial/macrophage cells | +++ | + | ++ |
ER Stress proteins, UPS and cell receptors | |||||
HSP70 | Heat shock protein 70 (polyclonal, 1:1,000; Dako) | Punctate deposits in arteriolar wall | +++ | + | + |
HSP27 | Heat shock protein 27 (polyclonal, 1:1,000; Dako) | Degenerated VSMC in media | +++ | + | + |
GRP78 | Glucose regulated protein 78 (polyclonal, 1:1,000; Dako) | Degenerated VSMC in media | +++ | ++ | ++ |
ERp78 | ER stress-related proteins (CHOP and BIP/ERP78) (polyclonal, 1:100; Abcam) | Punctate deposits in arteriolar wall | +++ | + | +++ |
Calnexin | ER stress marker (polyclonal, 1:100, AbCam) | Punctate deposits in arteriolar wall | +++ | + | +++ |
Ubiquitin | Ubiquitin protease system (UPS) (polyclonal, 1:50; Dako) | Degenerated VSMC in media | ++ | + | ++ |
ICAM-1 | Intercellular Adhesion Molecule-1, CD54 (polyclonal, 1:300; Dako) | Reactivity in arteries and perivascular region | +++ | + | ++ |
Cleaved Caspase 3 | Caspase-3 (polyclonal, 1:100, Cell Signalling Technology) | Punctate deposits in arteriolar wall | +++ | ++ | +++ |
Serum Amyloid Protein | SAP (polyclonal, 1:1000; Dako) | Diffuse reactivity in arteries | +++ | + | ++ |
Cystatin C (CST3) | CST3 (polyclonal, 1:1000, Dako) | Diffuse reactivity in arteries | +++ | + | ++ |
NOTCH3 Signalling* | |||||
N3ECD | A1-1 ab to NOTCH3 extracellular domain (polyclonal, 1:2,500; NILS [65] | ↑N3ECD deposition | 85.8 (4.9) | 0 | 0 |
N3ICD | C2 ab to NOCTCH3 intracellular domain (polyclonal, 1:1,000 ; NILS [31] | ↔N3ICD reactivity | 85.4 (2.3) | 82.3 (8.0) | 80 (5.0) |
Complement System | |||||
C1q | Complement C1q (polyclonal, 1:1,000; Dako) | Â | 0 | 0 | 0 |
C3d | Complement 3 (polyclonal, 1:1,000; Dako) | Â | 75.6 (5.2) | 0 | 0 |
CFB | Complement Factor B (polyclonal, 1:1,000; Serotec) | Â | 69.8 (6.3) | 0 | 0 |
C5-9 | Complement C5-9 complex (polyclonal, 1:1,000; Dako) | Â | 63.9 (2.7) | 0 | 0 |