Association of viral load and integration status of HPV and cytokine profile in situ in women with low-grade intraepithelial lesion, high grade epithelial lesion and invasive squamous cell carcinoma.

Abstract

Introduction: Infection with oncogenic types of human papillomavirus (HPV) is the most important factor for development of intraepithelial lesions and invasive squamous cell carcinoma. The immune response has an important role in the natural history of HPV infection in uterine cervix, and the cytokines are molecules that regulate the viral transcription. Objectve: To evaluate the pattern of cytokines in cervical biopsies of women with low-grade squamous intraepithelial lesions (LSIL), high-grade intraepithelial lesion (HSIL) and invasive squamous cell carcinoma (EC), and associate that pattern with the viral load and integration status of HPV. Patients and Methods: Will be included in the study 120 women with clinical and histological diagnosis of LSIL (n = 40), HSIL (n = 40), EC (n = 40) and 40 women with suspected of HPV-induced disease, but no pathological changes on cervix biopsy with a mean age paired with women in the study group (control group) attending in the Colposcopy Clinic of the Botucatu Medical School (UNESP) and in the Preventive Gynecology, Hospital Amaral Carvalho, Jaú, SP. The pathological examinations are performed in the Department of Pathology, Faculty of Medicine of Botucatu (UNESP). The search for HPV in the fragments of cervical biopsies obtained from colposcopic examination will be performed using the technique of polymerase chain reaction (PCR) using specific primers (MY9/11 and GP5+/GP6+) and genotyping will be performed by technique Linear Array HPV (Roche Molecular Diagnostics). The search for viral load and integration status of HPV DNA will be evaluated by PCR in real time. For the evaluation of immunostaining of cytokines of interest (IL-1², IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IFN-gama and TNF-alfa)) in fragments of cervical biopsies will be used the technique of immunohistochemistry. Data analysis will be conducted in accordance with the statistics, considering the results obtained.