Evaluation of Serum IL‑40 and IL‑41 as Integrated Immunometabolic Biomarkers in Iraqi women with Breast Cancer: A Case – Control Study

Abstract

Background: Breast cancer (BC) is a complex malignancy characterized by the uncontrolled proliferation of the mammary epithelium. Emerging research highlights the pivotal role of novel cytokines in the tumor microenvironment. Specifically, the recently identified immunomodulators, Interleukin-40 (IL-40) and Interleukin-41 (IL-41), have been shown to orchestrate B-cell biology, chronic inflammation, and metabolic homeostasis. These cytokines represent critical mediators that influence the intricate pathological landscape and progression of various malignancies, including breast cancer.

Objective: This study aims to quantify serum IL-40 and IL-41 levels in Iraqi women diagnosed with breast cancer and to evaluate their association with clinic-pathological features, disease staging and explore the potential of these cytokines as robust diagnostic and prognostic biomarkers within the clinical setting.

Materials and Methods: This case-control study involved 180 Iraqi women, aged 45–60 years. The cohort consisted of 90 newly diagnosed breast cancer patients at various clinical stages and 90 age-matched healthy individuals who served as the control group. Venous blood samples were collected from all participants to obtain serum. Serum concentrations of IL-40 and IL-41 were quantified using commercially available enzyme-linked immunosorbent assay (ELISA) kits. Additionally, lipid profiles and liver functional enzymes were assessed via spectrophotometric analysis.

Result: The study demonstrated that serum IL-40 and IL-41 levels were significantly elevated in breast cancer patients compared to healthy controls (P < 0.001), with high concentrations persisting across all clinical stages. Furthermore, patients exhibited a distinct pro-inflammatory and metabolic profile characterized by significant dyslipidemia (increased TC, TG, LDL-C, and VLDL-C), hepatic dysfunction (elevated ALT, AST, and ALP), and significantly reduced HDL-C levels (P < 0.05). A strong positive correlation was observed between IL-40 and IL-41 (r = 0.90; P < 0.001). Both cytokines showed significant positive correlations with all measured parameters, except for a negative correlation with HDL-C. ROC curve analysis revealed that IL-40 and IL-41 possess high diagnostic potential, with sensitivities of 92.0% and 74.5%, specificities of 88.5% and 70.0%, and Area Under the Curve (AUC) values of 0.892 and 0.738, respectively (at cut-off values of 107.63 pg/mL and 116.35 pg/mL).

Conclusion: Serum levels of IL-40 and IL-41 are significantly elevated in breast cancer patients compared to healthy controls, demonstrating a robust association with disease stage and severity. These findings strongly suggest that the IL-40 and IL-41 axis serves as a promising panel of non-invasive biomarkers for the early diagnosis and prognostic monitoring of breast cancer.