Assessment of the Mechanisms of Antiproliferative Therapy for Severe Cervical Intraephelial Neoplasia

Aim. The aim of the present study was to evaluate the changes in cytokine factors in cervical secretions after antiviral and immunomodulatory treatment in patients infected with the human papillomavirus (HPV). Design. Prospective randomized study. Materials and methods. Fifteen female patients aged 18 to 49 years, infected with HPV of high carcinogenic risk (mainly types 16 and 18), with histologically confirmed severe cervical intraepithelial neoplasia were included in the study. Patients received inosin pranobex orally 1000 mg 3 times a day for 10 days at 10-day intervals for 3 courses and 3,3'-diindolylmethane intravaginally as suppositories 100 mg 2 times a day for 3 months. For all study participants were used the polymerase chain reaction method (PCR) to test HPV and cervical mucus sampling before and after treatment. The content of cytokines in cervical mucus was determined using a commercial Bio-Plex Pro Human Cytokine 27-plex Assay Bio-Plex Pro™ kit manufactured by Bio-Rad Laboratories Inc. (USA) on a Luminex 200 analyzer (Luminex Corporation, USA) according to the manufacturer's recommendations. Ratio of concentrations in cervical mucus of interleukin (IL)-10/IL-17A was calculated as immunoregulatory index IL-10/IL-17A. Correlation analysis was performed between vascular endothelial growth factor (VEGF) and IL-10 IL-17A. Statistical processing was performed using Excel 365 and SPSS 20.0 software package. Differences were considered statistically significant (the null-hypothesis was rejected) at p < 0.05. Results. After the therapy in the general group of patients, a decrease in cytokines IL-17A and IL-1ra, chemokines IL-8, interferon-γ inducible protein 10 (IP-10) and monocyte chemotactic protein-1 (MCP-1), as well as growth factors: platelet-derived growth factor BB (PDGF-BB), VEGF and granulocyte-macrophage colony-stimulating factor (G-CSF). Statistically significant decreasing trend after the end of treatment (0.05 < p < 0.1) was found for IL-17A, G-CSF, MCP-1 and PDGF-BB, while the IL-10/IL-17A ratio increased. In a subgroup of 10 patients (62%) with a final negative HPV test, a statistically significant (p < 0.05) decrease in mucus IL-17A, IL-1ra and VEGF was observed. A decreasing trend (0.05 < p < 0.1) was found for G-CSF. In 5 (38%) HPV-positive patients statistically significantly (p < 0.05) decreased the concentrations of IL-8 and VEGF in cervical mucus. There was a tendency to decrease (0.05 < p < 0.1) the MCP-1 content. With little change in level of IL-10, the IL-10/IL-17A immunoregulatory index increased after treatment not only in the total group, but also in samples of participants with both negative and positive tests for HPV infection. In addition, treatment led to a decrease in VEGF and a positive correlation (r = 0.41; p < 0.05) was found between changes in VEGF and IL-17A concentrations. Conclusion. Complex administration of inosine pranobex orally with 3,3'-diindolylmethane topically have immunomodulatory, antiinflammatory, antiviral and, possibly, antiangiogenic and antiproliferative effects. Determination of VEGF, IL-10 and IL-17A content in cervical mucus followed by calculation of the immunoregulatory index of IL-10/IL-17A could be useful for prognosis and evaluation of treatment efficacy of HPV-induced cervical mucosal dysplasia. Keywords: cervical intraepithelial dysplasia, cytokine profile, cervical mucus, immunomodulatory therapy.