HBeAg status, cytokine profile, and microRNA-122 predict fibrosis in chronic hepatitis B

Aim: The natural history of chronic hepatitis B (CHB) is governed by hepatitis B e-antigen (HBeAg) status, viral replication, and host immunity. Reliable non-invasive markers of hepatic fibrosis and necroinflammation remain limited. The present study aimed to quantify serum interleukin-6 (IL-6), IL-8, IL-10, interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), transforming growth factor-beta (TGF-β), and microRNA-122 (miR-122) across stages of fibro-inflammatory progression in CHB, evaluate their association with HBeAg status and hepatic injury, and compare their diagnostic accuracy with conventional non-invasive indices. Methods: In a cross-sectional case-control study, 90 CHB patients and 100 healthy controls were enrolled at three Iraqi tertiary centers (November 2024–June 2025). Biomarkers were quantified and correlated with HBeAg status and liver stiffness measured by transient elastography (FibroScan), and compared with aspartate aminotransferase/platelet ratio index (APRI) and fibrosis-4 index (FIB-4). Results: Hepatitis B virus (HBV) DNA (6.5 ± 1.8 vs. 2.8 ± 1.2 log10 IU/mL; P < 0.001), the proportion with advanced fibrosis (F3–F4: 46% vs. 18%; P = 0.004), and liver stiffness (9.8 ± 5.2 vs. 7.5 ± 4.8 kPa; P = 0.04) were greater in HBeAg-positive than in HBeAg-negative patients (Figure 1; Table S1). IL-6, IL-8, TNF-α, and TGF-β increased stepwise with fibrosis severity, whereas IL-10 and IFN-γ followed a biphasic course. miR-122 was markedly downregulated versus controls (P < 0.001), declining progressively from F0 (0.0440-fold) to F4 (0.0002-fold). The composite panel (IL-6, TNF-α, miR-122) discriminated significant fibrosis (F ≥ 2) with an area under the receiver operating characteristic curve (AUC) of 0.93 [95% confidence interval (CI): 0.87–0.97], outperforming APRI and FIB-4. HBeAg positivity independently predicted advanced fibrosis [adjusted odds ratio (OR) = 4.2; 95% CI: 2.0–8.9]; miR-122 below 0.001-fold was associated with a 6.4-fold higher likelihood of advanced fibrosis (OR = 6.4, 95% CI: 2.8–14.5). Conclusions: Persistent HBeAg expression, elevated pro-inflammatory cytokines, and suppressed miR-122 constitute an integrated molecular signature of fibro-inflammatory severity in CHB. This multi-marker panel offers an accurate, accessible, non-invasive tool for fibrosis risk stratification, particularly in resource-limited settings.