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The severity of experimental arthritis is independent of IL-36 receptor signaling.

Lamacchia C, Palmer G, Rodriguez E, Martin P, Vigne S, Seemayer CA, Talabot-Ayer D, Towne JE, Gabay C - Arthritis Res. Ther. (2013)

Bottom Line: IL-36 exerts proinflammatory effects in skin and lung and stimulates T cell responses.As opposed to anti-IL-1RI antibody treatment, the injection of an anti-IL-36R antibody was devoid of effect on the development and severity of CIA.The severity of joint inflammation and structural damage in AIA was also unaltered by anti-IL-36R antibody treatment.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: Interleukin (IL)-36 refers to three related IL-1 family cytokines, IL-36α, IL-36β, and IL-36γ, that bind to the IL-36 receptor (IL-36R). IL-36 exerts proinflammatory effects in skin and lung and stimulates T cell responses. In the present study, we examined the expression and function of IL-36R and its ligands in experimental arthritis.

Methods: Collagen-induced arthritis (CIA), antigen-induced arthritis (AIA), and K/BxN serum transfer-induced arthritis were induced according to standard protocols. Messenger RNA levels for IL-36R and its ligands in the joints of mice with CIA were determined by RT-qPCR. Mice with CIA were injected with a blocking monoclonal anti-IL-36R, a blocking anti-IL-1RI, or their isotype-matched control antibodies at the time of arthritis onset. Anti-IL-36R or control antibodies were also injected at the time of AIA induction. Finally, IL-36R-deficient mice were examined in AIA and serum transfer-induced arthritis. The development and severity of arthritis were assessed by clinical and histological scoring.

Results: IL-36R, IL-36Ra and IL-36γ mRNA were detected in the joints of mice with CIA, but their levels did not correlate with arthritis severity. As opposed to anti-IL-1RI antibody treatment, the injection of an anti-IL-36R antibody was devoid of effect on the development and severity of CIA. The severity of joint inflammation and structural damage in AIA was also unaltered by anti-IL-36R antibody treatment. Finally, the severity of AIA and K/BxN serum transfer-induced arthritis was similar in IL-36R-deficient and wild-type mice.

Conclusions: The development and severity of experimental arthritis are independent of IL-36R signaling.

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Treatment with a monoclonal anti-IL-36 receptor (R) antibody does not modify the course of collagen-induced arthritis (CIA). CII-immunized mice (n = 10/group) were treated with anti-IL36R (M616, squares, white columns), anti-IL-1RI (M147, triangles, dashed columns) or isotype-matched control antibodies (4G8 (circles, grey columns) for M616, 4D2 (triangles, black columns) for M147), as described in Materials and methods. Results show the incidence of arthritis (A), the clinical severity of articular inflammation (B) and the number of arthritic paws (C) on days 21 to 41. The systemic inflammatory response (D) was assessed by measuring circulating IL-6 levels on day 41 after the first immunization. Joint sections from all mice were evaluated on day 41 for histological scores (E) and histological features (F). All sections were scored for inflammation, cartilage erosion and neutrophil infiltration. Images are representative of H&E- or toluidine blue-stained sections of knee joints for each group (original magnification × 10). (G) Levels of CXCL-1 (ng/mL) were determined by ELISA in ankle extracts of each mouse on day 41 and normalized by the total protein concentration (mg/mL). (B, C, E) Values are the mean ± standard error of the mean. ***P < 0.001, anti-IL-1RI versus isotype control 4D2; &P < 0.05 and &&P < 0.01, anti-IL-1RI versus anti-IL-36R, as assessed by Kruskal-Wallis test (B and C) or by analysis of variance (ANOVA), followed by unpaired two-tailed Student's t-test (E). (D, G) Results are shown as individual values for each mouse (symbols) and mean values (lines) **P < 0.01, anti-IL-1RI versus isotype control 4D2, as assessed by ANOVA, followed by unpaired two-tailed Student's t-test.
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Figure 3: Treatment with a monoclonal anti-IL-36 receptor (R) antibody does not modify the course of collagen-induced arthritis (CIA). CII-immunized mice (n = 10/group) were treated with anti-IL36R (M616, squares, white columns), anti-IL-1RI (M147, triangles, dashed columns) or isotype-matched control antibodies (4G8 (circles, grey columns) for M616, 4D2 (triangles, black columns) for M147), as described in Materials and methods. Results show the incidence of arthritis (A), the clinical severity of articular inflammation (B) and the number of arthritic paws (C) on days 21 to 41. The systemic inflammatory response (D) was assessed by measuring circulating IL-6 levels on day 41 after the first immunization. Joint sections from all mice were evaluated on day 41 for histological scores (E) and histological features (F). All sections were scored for inflammation, cartilage erosion and neutrophil infiltration. Images are representative of H&E- or toluidine blue-stained sections of knee joints for each group (original magnification × 10). (G) Levels of CXCL-1 (ng/mL) were determined by ELISA in ankle extracts of each mouse on day 41 and normalized by the total protein concentration (mg/mL). (B, C, E) Values are the mean ± standard error of the mean. ***P < 0.001, anti-IL-1RI versus isotype control 4D2; &P < 0.05 and &&P < 0.01, anti-IL-1RI versus anti-IL-36R, as assessed by Kruskal-Wallis test (B and C) or by analysis of variance (ANOVA), followed by unpaired two-tailed Student's t-test (E). (D, G) Results are shown as individual values for each mouse (symbols) and mean values (lines) **P < 0.01, anti-IL-1RI versus isotype control 4D2, as assessed by ANOVA, followed by unpaired two-tailed Student's t-test.

Mentions: Consecutively, we tested the role of IL-36R signaling in CIA using DBA/1 mice. We compared the effect of an i.p. injected anti-IL-36R antibody, administered at disease onset, with that of an isotype-matched control antibody. An additional group of DBA/1 mice served a positive control and was treated with a blocking anti-IL-1RI antibody to compare the relative contributions of IL-36R ligands and IL-1 cytokines (IL-1α and IL-1β) to the development and severity of CIA. Treatment of mice with the blocking anti-IL-36R antibody was devoid of effect on the incidence (Figure 3A) or severity of CIA (Figure 3B and 3C). In contrast, inhibition of IL-1RI signaling resulted in marked attenuation of the disease. Consistent with these results, histological examination at the end of the experiment showed a complete protection in mice treated with the anti-IL-1RI antibody, whereas articular inflammation and structural damage were similar in mice injected with the anti-IL-36R or with isotype-matched control antibodies (Figure 3E and 3F). Similarly, serum IL-6 levels were elevated in mice treated with the anti-IL-36R antibody or with isotype-matched control antibodies, while circulating IL-6 was undetectable in anti-IL-1RI antibody-treated mice (Figure 3D). Finally, the levels of different inflammatory mediators, such as IL-6 and CXCL-1, were elevated in the joints of anti-IL-36R antibody and isotype control antibody-treated mice, while they were drastically decreased in mice treated with the anti-IL-1RI antibody (Figure 3G; data not shown). Taken together, our results suggest that there is no major contribution of IL-36R ligands to the development and severity of CIA.


The severity of experimental arthritis is independent of IL-36 receptor signaling.

Lamacchia C, Palmer G, Rodriguez E, Martin P, Vigne S, Seemayer CA, Talabot-Ayer D, Towne JE, Gabay C - Arthritis Res. Ther. (2013)

Treatment with a monoclonal anti-IL-36 receptor (R) antibody does not modify the course of collagen-induced arthritis (CIA). CII-immunized mice (n = 10/group) were treated with anti-IL36R (M616, squares, white columns), anti-IL-1RI (M147, triangles, dashed columns) or isotype-matched control antibodies (4G8 (circles, grey columns) for M616, 4D2 (triangles, black columns) for M147), as described in Materials and methods. Results show the incidence of arthritis (A), the clinical severity of articular inflammation (B) and the number of arthritic paws (C) on days 21 to 41. The systemic inflammatory response (D) was assessed by measuring circulating IL-6 levels on day 41 after the first immunization. Joint sections from all mice were evaluated on day 41 for histological scores (E) and histological features (F). All sections were scored for inflammation, cartilage erosion and neutrophil infiltration. Images are representative of H&E- or toluidine blue-stained sections of knee joints for each group (original magnification × 10). (G) Levels of CXCL-1 (ng/mL) were determined by ELISA in ankle extracts of each mouse on day 41 and normalized by the total protein concentration (mg/mL). (B, C, E) Values are the mean ± standard error of the mean. ***P < 0.001, anti-IL-1RI versus isotype control 4D2; &P < 0.05 and &&P < 0.01, anti-IL-1RI versus anti-IL-36R, as assessed by Kruskal-Wallis test (B and C) or by analysis of variance (ANOVA), followed by unpaired two-tailed Student's t-test (E). (D, G) Results are shown as individual values for each mouse (symbols) and mean values (lines) **P < 0.01, anti-IL-1RI versus isotype control 4D2, as assessed by ANOVA, followed by unpaired two-tailed Student's t-test.
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Figure 3: Treatment with a monoclonal anti-IL-36 receptor (R) antibody does not modify the course of collagen-induced arthritis (CIA). CII-immunized mice (n = 10/group) were treated with anti-IL36R (M616, squares, white columns), anti-IL-1RI (M147, triangles, dashed columns) or isotype-matched control antibodies (4G8 (circles, grey columns) for M616, 4D2 (triangles, black columns) for M147), as described in Materials and methods. Results show the incidence of arthritis (A), the clinical severity of articular inflammation (B) and the number of arthritic paws (C) on days 21 to 41. The systemic inflammatory response (D) was assessed by measuring circulating IL-6 levels on day 41 after the first immunization. Joint sections from all mice were evaluated on day 41 for histological scores (E) and histological features (F). All sections were scored for inflammation, cartilage erosion and neutrophil infiltration. Images are representative of H&E- or toluidine blue-stained sections of knee joints for each group (original magnification × 10). (G) Levels of CXCL-1 (ng/mL) were determined by ELISA in ankle extracts of each mouse on day 41 and normalized by the total protein concentration (mg/mL). (B, C, E) Values are the mean ± standard error of the mean. ***P < 0.001, anti-IL-1RI versus isotype control 4D2; &P < 0.05 and &&P < 0.01, anti-IL-1RI versus anti-IL-36R, as assessed by Kruskal-Wallis test (B and C) or by analysis of variance (ANOVA), followed by unpaired two-tailed Student's t-test (E). (D, G) Results are shown as individual values for each mouse (symbols) and mean values (lines) **P < 0.01, anti-IL-1RI versus isotype control 4D2, as assessed by ANOVA, followed by unpaired two-tailed Student's t-test.
Mentions: Consecutively, we tested the role of IL-36R signaling in CIA using DBA/1 mice. We compared the effect of an i.p. injected anti-IL-36R antibody, administered at disease onset, with that of an isotype-matched control antibody. An additional group of DBA/1 mice served a positive control and was treated with a blocking anti-IL-1RI antibody to compare the relative contributions of IL-36R ligands and IL-1 cytokines (IL-1α and IL-1β) to the development and severity of CIA. Treatment of mice with the blocking anti-IL-36R antibody was devoid of effect on the incidence (Figure 3A) or severity of CIA (Figure 3B and 3C). In contrast, inhibition of IL-1RI signaling resulted in marked attenuation of the disease. Consistent with these results, histological examination at the end of the experiment showed a complete protection in mice treated with the anti-IL-1RI antibody, whereas articular inflammation and structural damage were similar in mice injected with the anti-IL-36R or with isotype-matched control antibodies (Figure 3E and 3F). Similarly, serum IL-6 levels were elevated in mice treated with the anti-IL-36R antibody or with isotype-matched control antibodies, while circulating IL-6 was undetectable in anti-IL-1RI antibody-treated mice (Figure 3D). Finally, the levels of different inflammatory mediators, such as IL-6 and CXCL-1, were elevated in the joints of anti-IL-36R antibody and isotype control antibody-treated mice, while they were drastically decreased in mice treated with the anti-IL-1RI antibody (Figure 3G; data not shown). Taken together, our results suggest that there is no major contribution of IL-36R ligands to the development and severity of CIA.

Bottom Line: IL-36 exerts proinflammatory effects in skin and lung and stimulates T cell responses.As opposed to anti-IL-1RI antibody treatment, the injection of an anti-IL-36R antibody was devoid of effect on the development and severity of CIA.The severity of joint inflammation and structural damage in AIA was also unaltered by anti-IL-36R antibody treatment.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: Interleukin (IL)-36 refers to three related IL-1 family cytokines, IL-36α, IL-36β, and IL-36γ, that bind to the IL-36 receptor (IL-36R). IL-36 exerts proinflammatory effects in skin and lung and stimulates T cell responses. In the present study, we examined the expression and function of IL-36R and its ligands in experimental arthritis.

Methods: Collagen-induced arthritis (CIA), antigen-induced arthritis (AIA), and K/BxN serum transfer-induced arthritis were induced according to standard protocols. Messenger RNA levels for IL-36R and its ligands in the joints of mice with CIA were determined by RT-qPCR. Mice with CIA were injected with a blocking monoclonal anti-IL-36R, a blocking anti-IL-1RI, or their isotype-matched control antibodies at the time of arthritis onset. Anti-IL-36R or control antibodies were also injected at the time of AIA induction. Finally, IL-36R-deficient mice were examined in AIA and serum transfer-induced arthritis. The development and severity of arthritis were assessed by clinical and histological scoring.

Results: IL-36R, IL-36Ra and IL-36γ mRNA were detected in the joints of mice with CIA, but their levels did not correlate with arthritis severity. As opposed to anti-IL-1RI antibody treatment, the injection of an anti-IL-36R antibody was devoid of effect on the development and severity of CIA. The severity of joint inflammation and structural damage in AIA was also unaltered by anti-IL-36R antibody treatment. Finally, the severity of AIA and K/BxN serum transfer-induced arthritis was similar in IL-36R-deficient and wild-type mice.

Conclusions: The development and severity of experimental arthritis are independent of IL-36R signaling.

Show MeSH
Related in: MedlinePlus