Soluble CD40 Levels in Rheumatoid Arthritis: Association with Serum Cytokines and Autoantibody Status

Macrophage polarization induced immune dysregulation in rheumatoid arthritis

Background and objective: Rheumatoid arthritis (RA) is one of the most prominent inflammatory autoimmune illnesses, causing polyarticular synovitis. The precise causes of RA are still unknown; however, chronic inflammation patterns are influenced by a combination of immunological, environmental, and epigenetic factors. Anti-CCP antibodies are a particular biomarker for RA diagnosis and severity. Alterations of complement components and pro- and anti-inflammatory cytokines are critical to the pathogenesis of RA. The current study intended to compare and examine the immunological properties of anti-CCP, IL-10, IL-17, and C5a antibodies in samples from RA patients and healthy individuals. The study evaluated medication exposure and various immune markers simultaneously to enhance immunological profiling, clinical accuracy, and classification of patients with RA Methods: The Cobas e 411 analyzer was used to measure the anti-CCP antibody levels in the serum of 60 RA patients and 40 healthy individuals. Enzyme-linked immunosorbent assay (ELISA) technique from the BioTek ELx800 was used to estimate the levels of IL-17, IL-10, and C5a in the sera of participants. Both groups were matched in age and ethnicity. An unpaired T-test was used to analyze data statistically using the GraphPad Prism 8 program. The ANOVA test was used for more than 2 groups. Results: The current study indicated that patients with rheumatoid arthritis (RA) had a significantly lower concentration of IL-10 serum levels as compared to the healthy controls, yet with increased levels of IL-17, C5a, and anti-CCP antibodies. Some of the causes of the decrease in IL-10 might be chronic inflammation and the suppressive action of the pro-inflammatory cytokines on its production and signaling. Further, RA patients under corticosteroid treatment had reduced levels of IL-17, C5a, and anti-CCP antibodies compared to untreated patients, showing corticosteroid therapy lowers the inflammatory mediators without restoring the IL-10 levels. Conclusion: It can be concluded that macrophages from rheumatoid arthritis patients shifted from anti-inflammatory (like IL-10) to pro-inflammatory (like IL-17) properties. Patients with rheumatoid arthritis had lower levels of anti-inflammatory cytokine markers like IL-10 and greater levels of pro-inflammatory cytokines like IL-17, inflammatory markers like C5a, and rheumatoid arthritis markers like anti-CCP.

Cytokine Profiles in Rheumatoid Arthritis: Relationship of TNF-α , IL-17, and IL-13 with Autoantibodies (RF and Anti-CCP) and Clinical Disease Severity

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by synovial inflammation and autoantibody production. This study aimed to assess serum levels of TNF-α , IL-17, and IL-13 in RA patients and to explore their association with disease severity and autoantibody status in comparison to healthy controls. The study involved 67 RA patients and 20 healthy individuals. Patients were initially classified based on autoantibody status (RF-positive vs. RF-negative and anti-CCP-positive vs. anti-CCP-negative), and further categorized into low, moderate, and high disease activity groups. Cytokine levels were measured by ELISA, CRP and RF by turbidimetry, and anti-CCP levels by electrochemiluminescence immunoassay. Disease severity was evaluated using DAS28-ESR. RA patients had significantly higher levels of TNF-α , IL-17, CRP, ESR, and autoantibodies compared to controls (p < 0.05). TNF-α levels were positively associated with RF and anti-CCP levels (p < 0.05). IL-17 levels showed no significant association with RF or anti-CCP status but were substantially elevated in moderate and severe RA stages. TNF-α levels were also higher in severe RA cases. IL-13 levels showed no significant difference between RA patients and controls and did not correlate with disease severity or autoantibody levels, though it was directly related to IL-17. RF demonstrated the highest diagnostic accuracy for RA (AUC = 0.905), followed by CRP (AUC = 0.779) and IL-17 (AUC = 0.753). TNF-α and IL-17 may be involved in the pathophysiology of RA, whereas RF, CRP, and IL-17 have potential diagnostic value. Further studies are required to clarify the role of IL-13 in RA.

Distinct serum chemokine signatures differentiate rheumatoid arthritis from non-rheumatoid arthritides

: Background: Early and accurate differential diagnosis of rheumatoid arthritis (RA) remains challenging, particularly in patients with seronegative or non-specific inflammatory arthritides. Because cytokines and chemokines play central roles in immune activation and leukocyte trafficking, we investigated whether serum cytokine and chemokine profiles could discriminate RA from non-rheumatoid arthritides (NRA) and healthy controls (HCs). Methods: One hundred and fifty-seven RA patients (104 anti-cyclic citrullinated peptide antibody (ACPA)-positive and 53 ACPA-negative patients), 33 NRA patients and 86 HCs were included. Fourteen cytokines and twelve chemokines were measured in the sera of the above three cohorts, using high-sensitivity chemiluminescence immunoassay. Results were analyzed by non-parametric Mann-Whitney U-test and multiple logistic regression analysis. Results: Among the parameters, eight chemokines (CCL3, 4, 11, 20, 27, CXCL9, 10 and 13) were significantly upregulated in RA patients as compared with NRA patients and HCs (Mann-Whitney U-test: p < 0.01). Multiple logistic regression analysis revealed that the combination of four chemokines (CCL3, 11, 27 and CXCL13) can effectively discriminate RA from NRA patients and HCs (Area Under the Curve (AUC) = 0.92), as well as ACPA-negative RA from NRA patients (AUC = 0.73). Conclusions: A serum four-chemokine signature consisting of CCL3, CCL11, CCL27, and CXCL13 discriminated RA, including ACPA-negative RA, from non-rheumatoid arthritides and healthy controls. These findings suggest that serum chemokine profiling may provide adjunctive diagnostic information for the differential diagnosis of RA.

Dissociation between systemic inflammation and symptom burden in rheumatoid arthritis: biomarker – clinical discordance in an African population

The relationship between systemic inflammation, autoimmunity, and patient-reported outcomes in individuals with rheumatoid arthritis (RA) remains poorly understood, particularly in African communities experiencing delayed diagnoses and limited access to biological treatments. This study aimed to evaluate the relationships among circulating inflammatory biomarkers, disease activity, and patient-reported outcomes in a cohort of individuals attending Kenyatta National Hospital in Kenya. We conducted a case–control study involving 44 patients with RA and 44 healthy controls at the Kenyatta National Hospital. We measured plasma levels of cytokines (tumour necrosis factor alpha (TNF-α), interleukin 6 (IL-6), and interleukin 1 beta (IL-1)), autoantibodies anti-citrullinated protein antibodies (ACPA) and rheumatoid factor (RF), and acute-phase reactants C-reactive protein (CRP) using standard enzyme-linked immunosorbent assay (ELISA) kits and erythrocyte sedimentation rate (ESR). Disease activity was assessed using the disease activity score-28 with erythrocyte sedimentation rate (DAS28-ESR), while patient-reported outcomes were captured using the pain visual analogue scale, health assessment questionnaire, and duration of morning stiffness. Statistical analyses involved Spearman’s correlations, receiver operating characteristic (ROC) curves, and multiple linear regression. All biomarker levels (ACPA, CRP, RF, IL-1β, IL-6, and TNF-α) were significantly higher in patients with RA than in controls (p < 0.001). Strong correlations were observed between inflammatory biomarkers (ACPA-CRP ρ = 0.70) and joint counts (ACPA-tender joints ρ = 0.56). ESR demonstrated the greatest discriminative ability for both high disease activity (AUC = 0.778) and severe pain (AUC = 0.775), whereas cytokines performed poorly (AUC < 0.60). Multiple regression revealed that ESR (β = 0.67) and tender joint count (β = 0.54) were the strongest predictors of DAS28-ESR (R2 = 0.92). This study revealed a significant dissociation between systemic inflammation and patient-reported symptoms in individuals with RA. Although biomarkers, such as ESR, autoantibodies, and cytokines, effectively reflect inflammatory burden and disease activity, they are poorly correlated with pain and functional outcomes. These findings highlight the multifactorial nature of RA symptoms and support the need for integrated assessment strategies that combine objective biomarkers with patient-reported measures to guide comprehensive therapeutic approaches. Study limitations included a modest sample size and a predominantly late-stage disease cohort.

Serum IL-22, miRNA-146a Expression and Total Antioxidant Status as Integrated Biomarkers in Rheumatoid Arthritis

Background: Rheumatoid Arthritis (RA) is a chronic systemic autoimmune disease characterized by persistent inflammation of the synovial joints. Over time, this inflammatory process contributes to cartilage destruction, bone erosion, functional impairment and several extra-articular complications [1,2]. Current evidence indicates that RA pathogenesis is not driven by a single mechanism, but rather by a complex interaction among cytokine dysregulation, epigenetic changes and oxidative stress. Objective: This study was designed to organize, analyze and interpret the available findings concerning serum IL-22, miRNA-146a relative expression and total antioxidant status in patients with RA compared with healthy controls, with particular attention to their biological significance and potential diagnostic value. Methods: A case-control dataset including 40 RA patients and 40 controls was analyzed. The previously excluded immunological variables were removed from the analysis according to the revised study plan.IL-22 was detected by ELISA Technique, whereas miRNA was restricted by qPCR and spectrophotometer was used to estimate total antioxidant. Statistical analysis was performed in an SPSS-style framework using descriptive statistics, independent-samples comparisons, Spearman correlation and Kruskal-Wallis testing where appropriate. Results: Patients with RA exhibited increased levels of all three investigated biomarkers when compared with the control group. Among these markers, miRNA-146a showed the greatest relative change, followed by total antioxidant status and IL-22. The reported ROC analysis indicated excellent diagnostic performance for miRNA-146a and total antioxidant status, whereas IL-22 demonstrated only moderate to borderline discriminatory ability. Conclusion: The available data suggest that miRNA-146a, IL-22 and total antioxidant status may collectively reflect an interconnected epigenetic, inflammatory and redox-related profile in RA. Among the studied biomarkers, miRNA-146a appeared to be the most informative indicator in the supplied dataset. However, further confirmation using original participant-level data and an independent validation cohort is still required before final journal submission.

Clinical and molecular classification of very early arthritis patients