1. by Joanna Shepherd, and Martin J.H. Nicklin
Elastic-Vessel Arteritis in Interleukin-1 Receptor Antagonist–Deficient Mice Involves Effector Th1 Cells and Requires Interleukin-1 Receptor
by Joanna Shepherd, and Martin J.H. Nicklin
Circulation
Volume 111(23):
June 14, 2005
Copyright © American Heart Association, Inc. All rights reserved.

2. Figure 1. Severity of aortic root inflammation in Sf3 as function of time.
Figure 1. Severity of aortic root inflammation in Sf3 as function of time. Severity class score in individual mice at age of killing. ○ indicates Il1rn−/− Sf3 mice; +, wild-type Sf3 mice.
Joanna Shepherd, and Martin J.H. Nicklin Circulation. 2005;111:
Copyright © American Heart Association, Inc. All rights reserved.

3. Figure 2. Graded infiltration by IFN-γ+ cells at aortic root.
Figure 2. Graded infiltration by IFN-γ+ cells at aortic root. Frozen sections were stained for antigen (brown) and counterstained with hematoxylin. A indicates aortic lumen. Scale bars are 200 μm. IFN-γ+ cells were stained brown in unaffected (grade 0) aorta from 151-day-old wild-type Sf3 female mouse (a); grade 1 infiltration in 24-day-old Il1rn+/+ Sf3 male (b; arrows indicate infiltration), grade 2 lesion from 146-day-old Il1rn−/− Sf3 male mouse (c), and grade 3 lesion from 155-day-old Il1rn−/− BALB/c male mouse (d). e–g, Detection of infiltrating cells at base of aortic valve of same mouse as Figure 2b. Serial sections were stained for CD4 (e), IFN-γ (f), and IL-4 (g).
Joanna Shepherd, and Martin J.H. Nicklin Circulation. 2005;111:
Copyright © American Heart Association, Inc. All rights reserved.

4. Figure 3. Vascular activation and cellular infiltration.
Figure 3. Vascular activation and cellular infiltration. Methods were as in Figure 1 except for Figure 3e. Scale bars are 200 μm. a and b, Microvasculature of unaffected 24-day-old female Il1rn−/− Sf3 mouse. a, E-selectin; b, CD31; c–e, Vascular activation in 179-day-old female Sf3 Il1rn−/− mouse. c, E-selectin (arrows indicate sites where endothelium appeared to be activated). d, Higher-resolution image from c. e, Elastin and van Gieson staining (arrow indicates margin of region of elastin destruction). A indicates aortic lumen.
Joanna Shepherd, and Martin J.H. Nicklin Circulation. 2005;111:
Copyright © American Heart Association, Inc. All rights reserved.

5. Figure 4. Modification of aortic root by inflammation.
Figure 4. Modification of aortic root by inflammation. Scale bars are 100 μm except where marked. a–c, Zinc-Tris fixed aortic root with neck of coronary artery in 175-day-old Il1rn−/− BALB/c male. a, Staining for MHC II. C indicates focus of inflammation at opening of coronary artery; E, elastic media. b, Section taken from <40 μm deeper than Figure 4a; elastin and van Gieson staining (EVG). c, Intermediate section stained with hematoxylin and eosin. C indicates focus of inflammation at opening of coronary artery. d, Coronary artery (C) in 175-day-old wild-type male BALB/c mouse; EVG. e, Higher magnification of d showing elastin (black). f, Cross section of entrance to coronary artery from deeper section but same block as panels a, b, and c; EVG. g, Adjacent section stained with hematoxylin and eosin. h, Higher magnification of Figure 4f. i–l, Frozen sections from seat of cusp of aortic valve in 155-day-old Il1rn−/− BALB/c male (also in Figures 5a through 5e). i, EVG; apparent destruction of elastin (black) in background of collagen (red) deposition. j, Enlargement of area of apparent elastin destruction in panel i. Small arrows indicate fine (new) elastin bundles; larger arrows indicate truncated fibers. k, Adjacent section stained for F4/80 (macrophages). Arrow indicates dense macrophage infiltration close to elastin destruction. Inset is approximately equivalent to Figure 5a through 5e. l, Direct UV-excited autofluorescence of matrix proteins in same section as panel k. Amorphous collagen creates pale background. Arrow shows macrophage cluster indicated in panel k. m, Cartilaginous nodule, stained with toluidine blue, at base of cusp of aortic valve in affected 207-day-old Il1rn−/− BALB/c×C57BL/6 F2 male. n, Aortic root of healthy 56-day-old Il1rn+/− Sf3 male, stained for F4/80. o, Same field showing UV-excited autofluorescence of elastin ribbons.
Joanna Shepherd, and Martin J.H. Nicklin Circulation. 2005;111:
Copyright © American Heart Association, Inc. All rights reserved.

6. Figure 5. Mediators of chronic inflammation.
Figure 5. Mediators of chronic inflammation. Serial frozen sections were stained for antigens as indicated. Images a through l are same scale (bars in a and m are 100 μm). a–e, Seat of aortic valve of 155-day-old male Il1rn−/− BALB/c mouse (also in Figures 4i through 4l). a, CD4. b, IL-4. c, IFN-γ. d, CD205; arrows indicate strongly positive cells. e, MHC II; arrows indicate morphologically dendritic cells. f–i, Serial frozen sections from 81-day-old male Il1rn−/− Sf3. Arrows indicate some areas positive for (f) chemokine CCL2 (MCP-1), (g) CD4, (h) E-selectin, and (i) CD31. j, Aortic root from 171-day-old wild-type BALB/c male stained for MHC II. Large arrow indicates ring of dendritic cells adjacent to media; small arrows show positive cells in valve matrix. k–m, Staining for CD4 and IFN-γ in serial sections from 221-day-old female Il1rn−/− mouse. Arrows indicate similar area. k, CD4. l, IFN-γ. m, Higher-resolution image costained black for IFN-γ (nickel-enhanced diaminobenzidine/peroxidase visualization) and red for CD4 (vector red/alkaline phosphatase visualization). For clarity, nuclei were not counterstained. Some costaining cell clusters are indicated.
Joanna Shepherd, and Martin J.H. Nicklin Circulation. 2005;111:
Copyright © American Heart Association, Inc. All rights reserved.