Cytokine Profiles in Cervical Intraepithelial Neoplasia: Associations with Lesion Severity and Exploratory Treatment...

Background: Persistent infection with high-risk human papillomavirus (hrHPV) is the principal cause of cervical intraepithelial neoplasia (CIN). Cytokines regulate antiviral immunity and tissue remodeling, and cytokine profiling may provide additional information about the biology of cervical lesions. Objective: To characterize cytokine profiles in cervical mucus and serum across grades of cervical intraepithelial neoplasia. Methods: The study included 90 women aged 18-80 years: 30 HPV-negative women with negative cytology (Control) and 60 HPV-positive women with histologically confirmed low-grade (LSIL, n = 30; Gr-1) or high-grade (HSIL, n = 30; Gr-2) squamous intraepithelial lesions. Participants were also stratified by minor (grade 1) or major (grade 2) colposcopic findings. An exploratory subgroup of 15 women with HPV16/18-positive HSIL received antiviral and immunomodulatory therapy. Concentrations of 27 cytokines (pg/mL)-FGF, eotaxin, G-CSF, GM-CSF, IFN-γ, IL-1ra, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12(p70), IL-13, IL-15, IL-17A, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF-BB, RANTES, TNF-α, and VEGF-were measured in serum and cervical mucus using the Bio-Plex Pro Human Cytokine 27-plex assay (Bio-Rad, USA) on a Luminex 200 platform. Statistical analyses were performed using SPSS version 23.0 and Microsoft Excel 365. Results are reported as medians and interquartile ranges [Q1-Q3]. A two-sided p-value < 0.05 was considered statistically significant. Results: Exploratory multivariate analyses identified IL-1β, IP-10, MCP-1, and FGF as the cytokines most strongly associated with lesion severity and colposcopic findings. In cervical mucus, HSIL was associated with the highest IL-1β concentration (0.83 pg/mL [0.18-4.91] vs. 0.30 pg/mL [0.04-0.69] in controls; p < 0.001) and the lowest IP-10 concentration (3.07 pg/mL [1.02-13.16] vs. 25.12 pg/mL [5.01-42.40]; p < 0.001). The highest mucus FGF concentration was observed in LSIL (1.73 pg/mL [1.33-2.27] vs. 0.32 pg/mL [0.17-0.59] in controls; p < 0.001), whereas MCP-1 was lower in LSIL than in HSIL (2.38 pg/mL [1.57-3.54] vs. 5.38 pg/mL [3.12-10.55]; p < 0.05). Similar patterns were observed in serum, except that IL-1β was lower than in controls. After treatment, HPV was undetectable in 10 women and remained detectable in 5. The overall cytokine profile changed only modestly. Among the five women with persistent HPV infection, cervical mucus MCP-1 was lower after treatment than before treatment (0.09 pg/mL [0.01-0.41] vs. 0.37 pg/mL [0.08-0.73]; p < 0.01). Conclusions: These findings show associations between lesion severity, colposcopic findings, and compartment-specific cytokine patterns. Because the study was cross-sectional and the treatment subgroup was small and uncontrolled, the results should be considered exploratory and require longitudinal validation.