Non-depleting anti-CD4 MAb prevents the onset of autoimmune arthritis.

<p>(A) Female SKG mice were immunized with 3mg curdlan i.p. together with 1 mg non-depleting anti-CD4 or an isotype control. The MAb was administered again on days 2 and 4. Anti-CD4 treated mice were protected from the development of autoimmune arthritis (n = 6, <i>P</i><0.001). Data, represented as mean ± SEM, are from two independent experiments. (B) Serum concentration of rheumatoid factor (RF) was measured by ELISA. Mice treated with anti-CD4 showed significantly lower levels of RF (n = 6, <i>P</i><0.001) (C) Histological sections stained with eosin-hematoxilin from the ankle and metatarso-phalangical joints from SKG mice in the absence and in the presence of anti-CD4 treatment, 90 days following curdlan immunization. (D) Serum concentration of IL-6, IFN-γ, TNF and MCP-1 in naive SKG mice, SKG mice exposed to curdlan, or curlan and anti-CD4. Naive SKG mice were age matched and did not develop arthritis in the absence of curdlan immunization (no induction). The serum levels of IL-6 (<i>P</i><0.05), IFN-γ (<i>P</i><0.01) and MCP-1 (<i>P</i><0.01) were significantly lower in anti-CD4 treated mice compared with animals injected with curldan in the absence of tolerizing MAbs. Differences in TNF concentration did not reach statistical significance. (E) The serum concentration of IL-10 and IL-17 in SKG mice exposed to curdlan, or curlan and anti-CD4 remained similar in the different experimental groups. Culture supernatants from Th17 cell culture were used as positive control.</p

Anti-CD4 MAb influences the local balance of Th17/Treg cells.

<p>(A) Frequency of splenic CD4⁺ T cells or CD4⁺Foxp3⁺ Treg cells from SKG mice exposed to curdlan, or curdlan + anti-CD4 treatment. No significant difference was observed between the two populations of animals. (B) Representative dot plots showing the frequency of splenic CD4⁺Foxp3⁺ T cells from SKG mice exposed to curdlan, or curdlan + anti-CD4 treatment. No significant difference was observed, as represented in panel A. (C) Foxp3 and IL-17 mRNA expression from the synovial membrane of SKG mice exposed to curdlan, or curdlan + anti-CD4 (mRNA expression levels were normalized to CD3 expression). (D) Frequency of Foxp3⁺ and IL-17⁺ T cells within draining LNs of SKG mice exposed to curdlan, or curdlan + anti-CD4.</p

CD4-blockade prevents Th17 polarization.

<p>(A) Sorted TCR-transgenic cells were stimulated <i>in vitro</i> with peptide-loaded DCs under culture conditions known to preferentially polarize Th17 cells, with the addition of recombinant TGF-β, IL-6, IL-1β, and anti-IFN-γ. After 5 days of culture we observed a significant reduction of IL-17⁺ cells in the presence of anti-CD4 (n = 6, <i>P</i><0.05). In contrast, anti-CD4 addition led to an increased frequency of Foxp3⁺ T cells (n = 6, <i>P</i><0.05). (B) Representative dot plots from the two different culture conditions. An independent experiment was performed with a peptide dose of 0.3 µM with similar results.</p

Vascular biomarkers and results of PAT assessment in SLE and RA, after controlling for baseline covariates.

<p>Results are presented as estimated marginal means (SE).</p>*<p>Adjusted for the following covariates: age, disease duration, total cholesterol, HDL, LDL, triglycerides, aspirin, hydroxychloroquine, methotrexate use, and prednisolone dose.</p>§<p>RHI and AIx results refer to 87 women with SLE and 75 with RA.</p><p>sICAM-1 – soluble intercellular adhesion molecule; sVCAM-1 – soluble vascular cell adhesion molecule; TM – thrombomodulin; TF – tissue factor; RHI – reactive hyperemia index; AIx – augmentation index.</p

Demographic and clinical characteristics of SLE and RA women.

<p>Results are presented as means (SD) or number of affected individuals and (%).</p><p>SLE – systemic lupus erythematosus; RA – rheumatoid arthritis; CV- cardiovascular; HDL – high density lipoprotein; LDL – low density lipoprotein, ns – non significant.</p

Vascular biomarkers and endothelial function in active disease and in remission.

<p>Results are expressed as estimated marginal means (SE) adjusted for prednisolone dose.</p>*<p>RHI and AIx results refer to 87 women with SLE and 75 with RA.</p><p>SLE – systemic lupus erythematosus; RA – rheumatoid arthritis; sICAM-1 – soluble inter-cellular adhesion molecule; sVCAM-1 – soluble vascular cell adhesion molecule; TM – thrombomodulin; TF – tissue factor; RHI – reactive hyperemia index; AIx – augmentation index.</p

Bone turnover markers quantification in control (N = 9) and arthritic rats (N = 13).

<p>Serum samples collected at day 22 (sacrificed) were analysed by ELISA technique. Bone resorption marker, CTX I (A) and bone formation marker, P1NP (B) were increased in arthritic rats (p = 0.0002 and p = 0.0010, respectively).</p

Calcium and Phosphorus bone content acquired by energy dispersive X-ray spectroscopy.

<p>Ca (A) and P (B) bone content were decreased in the arthritic group (N = 16) as compared with controls (N = 12). Bone powder was acquired from bone samples collected at day 22 post disease induction (sacrifice).</p

Bone histomorphometry assessment of the 4th lumbar vertebra (L4).

<p>Assessment of L4 in control (N = 12) and arthritic group (N = 16). (A) Illustrative Aniline blue stained sections of L4 vertebra collected at day 22 post disease induction (sacrifice). Bone volume per tissue volume or trabecular bone volume fraction (B) and trabecular thickness (C) were decreased in arthritic rats while trabecular separation (D) was increased. Magnification x12.5.</p

Mechanical analysis acquired by 3 point bending tests.

<p>Yield stress (A) and Ultimate stress (B) were decreased in arthritic rats (N = 16) as compared to controls (N = 12). Bone samples were collected at day 22 post disease induction (sacrifice).</p