Detection of immature or mature blood vessels in RA synovial tissues.

<p>Double immunoflurescent labeling of endothelium (CD31, red fluorescence) and pericytes/smooth muscle cells (aSMA, green fluorescence) in normal and RA synovial tissue is shown. Original magnification ×400. Right panels show the same area as in middle panels with higher magnification. Mature CD31+ vessels covered by aSMA+ periendothelial cells are marked by arrows, and immature CD31+ vessels lacking aSMA+ mural cells by arrow heads.</p

Mature and Immature Vessels in RA, OA or Normal Synovial Tissues.

<p>CD31+/aSMA+: Mature vessels; CD31+/aSMA–: Immature vessels; Total vessels represents the sum of both mature and immature vessels.</p><p>(*) RA versus OA.</p

Clinicopathological data stratified by the presence of Immature Vessels (IV).

<p>Data represent baseline data recorded at the time of biopsy. IV: immature CD31+/aSMA– vessels. LN: lymphoid neogenesis; DAS28: disease activity score; CRP: C-reactive protein; NS: Non-significant.</p><p>(*)IV- versus IV+ groups.</p><p>(†)RF or ACPA auto-antibodies.</p><p>(¶)p<0.05 but NS after correction for multiple testing.</p

Variation in the density of immature vessels stratified by the levels of response to anti-TNF-α therapy.

<p>Decrease in immature (left graphics) or mature (right graphics) vessels density between the first and second biopsy after anti-TNF-α therapy is shown stratified by EULAR responses: 0 = : No response; 1: Moderate response; 2: Good response. (*) Kruskall Wallys test and post hoc Dunns test (non-responders versus good responders).</p

Double labeling of lymphatic and CD31-positive vessels in RA synovial tissues.

<p>Lymphatic vessels were detected by immunoperoxidase (brown immunostaining) detection of podoplanin and double immunofluorescent labeling (red fluorescence) of CD31. The same field was photographed by light or fluorescent microscopy to show the position of CD31+ (arrowheads) and podoplanin+ vessels (arrows). Light microscopy image was inverted and merged with CD31 fluorescent image of the same field to show the relative position of podoplanin (blue) and CD31 (red) labeling.</p

Clinicopathological changes in patients after anti-TNFα therapy.

<p>CD31+/aSMA–: immature vessels. CD31+/aSMA+: mature vessels. p-value of basal versus post-anti-TNF values.</p><p>(*)Absolute decrease from basal values in patients achieving moderate or good EULAR response (responders) and non responders to anti-TNF.</p

Clinicopathological correlations of immature blood vessels in RA synovial tissues.

<p>Disease duration, DAS28 score, erosive disease, and synovial tissue infiltration by CD3, CD20 or CD68 cells is shown in groups with (+) or without (−) immature vessels as indicated. Density of mature or immature vessels in patients stratified by disease duration and levels of activity (low: DAS28<3.2, moderate 3.2–5.1, or high>5.1). Spearman's correlation coefficients between immature vessels density and disease duration, DAS28, CD3 or CD20 infiltration are shown. (*) p<0.05 (see text). ¶ p = 0.04 (Kruskall Wallys test and post hoc Dunns test (low versus moderate or high activity groups).</p

gp38 expression of cultured synovial fibroblasts.

<p>SF from RA, OA and normal synovial tissues, and normal skin dermal fibroblasts (DF) were cultured on glass coverslips and immunolabeled (red) for gp38 expression. TNF-α treated DF cultures are also shown (DAPI nuclear counterstaining). Flow cytometric detection of surface gp38 by in RA SF and DF untreated (basal) or treated with TNF-α for 24h. Data are representative of 6 SF and 3 DF lines (*p = 0.03 basal vs TNF-α treated). MFI: Mean fluorescence intensity.</p

Clinicopathological data of RA patients at biopsy.^*

<p>*Quantitative data are expressed as mean±SD and [range]. CRP, C-reactive protein; DAS28, 28-joint Disease Activity Score; DMARD, disease-modifying antirheumatic drug; ^†RF>30 IU/ml; anti-citrullinated protein antibodies ACPA>50 IU/ml.</p

Expression of gp38 in synovial tissues.

<p><b>(A)</b> IHC immunoperoxidase labelling of gp38 expression in normal, OA and RA tissues. RA CTRL: IgG1 isotype control. Lining and sublining areas, and a large lymphoid aggregate centered by a HEV (arrows) are shown. Bar: 50 µm. Original magnification 400x <b>(B)</b> Changes in gp38 expression after anti-TNF therapy. Representative imunoperoxidase labeling of gp38 of a single patient pre- and post-treatment. Bar: 100 µm. Original magnification 200x <b>(C)</b> Mean±SD gp38 immunostained fractional area in the different groups (OA vs RA Mann Whitney test *p<0.0001), and in basal (Pre) and post-anti-TNF-α therapy (Post) in 16 patients (RA Pre vs RA Post Wilcoxon matched-pairs signed rank test *p<0.0001).</p