Table 1.
Clinical characteristics of the study populations.
Figure 1.
Frequency of IL-17A+ and IL-17F + cells in SSc and morphea.
A. Immunohistochemical analysis of IL-17A and IL-17F in healthy (HD), lesional SSc and morphea skin. Arrows indicate IL-17A (upper panels) or IL-17F (lower panels) positive cells. Results are representative of 14 SSc, 5 Morphea and 8 HD individuals. Original magnification 20X, scale bar 100 µm. Insets, 2X. B. Frequency of IL-17A+ and IL-17F+ cells expressed as percentage of total cells. Each symbol represents a distinct individual and the line depicts the mean. Empty and full dots refer to limited and diffuse SSc, respectively. Significance was assessed by Mann-Whitney test.
Figure 2.
Presence of CD3+IL-17+ T cells and tryptase+IL17+ mast cells in morphea.
A. Indirect immunofluorescence analysis was used to assess the expression of IL-17A (green), in combination with CD3 (red, left panel) or Tryptase (red, right panel) and DAPI staining of nuclei (blue) in the dermis of a representative of 5 morphea biopsies. Original magnification 40X. B. Box-plots show the quantification of CD3+IL-17A+ and Tryptase+IL-17A+ cells expressed as percentage of total IL-17A+ cells in 4 HD, 4 SSc and 4 morphea skin section. The box represents values between 25th and 75th percentile with a line at the median (50th percentile). The whiskers extend above and below the box to show the highest and the lowest values.
Figure 3.
High frequency of IL-17E+ and low frequency of IL-17C + cells identifies a fibrosis-specific motif common to SSc and morphea.
A. Immunohistochemical analysis for IL-17C and IL-17E in healthy (HD), lesional SSc and morphea skin. Arrows indicate IL-17E (upper panels) or IL-17C (lower panels) positive staining. Results shown are representative of 14 SSc, 5 Morphea and 8 HD individuals. Original magnification 20X, scale bar 100 µm. Insets, 2X. B. Frequency of IL-17E+ and IL-17C+ cells expressed as percentage of total cells. Each symbol represents a distinct individual and the line depicts the mean. Empty and full dots refer to limited and diffuse SSc, respectively. Significance was assessed by Mann-Whitney test.
Figure 4.
Co-expression of IL-17A with IL-17C or IL-17E.
Indirect immunofluorescence analysis was used to assess the expression of IL-17A (green), in combination with IL-17C (A) or IL-17E (B) or IL-17F (C) (red), and DAPI staining for nuclei (blue) in the dermis of one biopsy of 3 assessed (1 HD, 1 SSc and 1 morphea). Original magnification 40X.
Figure 5.
IL-17A+ and IL-17F+ cell frequency positively correlates with disease duration.
IL-17A+ and IL-17F+ cells were identified by immunohistochemistry. Each symbol represents a single individual. Significance was assessed by Pearson's correlation test. mo = months.
Figure 6.
IL-17F and IL-17E, but not IL-17C, enhance MCP-1 and MMP-1 production by normal and SSc fibroblasts.
Dermal fibroblasts were cultured in triplicate in the presence of increasing amounts (3, 30, 300, 600 ng/ml) of IL-17C, IL-17E or IL-17F for 48 h. Box-plot show the levels of MCP-1, IL-6, IL-8, MMP-1 and type I collagen assessed in fibroblast culture supernatants from 4 HD and 4 SSc. The box represents values between 25th and 75th percentile with a line at the median (50th percentile). The whiskers extend above and below the box to show the highest and the lowest values. IL-17A (30 ng/ml), TNF (A, 1 ng/ml) or TGF-β (B, 10 ng/ml) were used as controls. Significance versus nil condition was assessed by one-sample t-test.