The Function Of Cytokines In The Cervical Immune Microenvironment With HR-HPV Persistent Infection

Objectives:To explore the changes of cervical local cytokines associated with Th1/Th2/Th17/Treg in the process of high risk human papillomavirus(HR-HPV)persistent infection.These cytokines are interleukin-2(IL-2)?IL-4?IL-6?IL-10??-interferon(IFN-?)?tumor necrosis factor(TNF)?IL-17 A and transforming growth factor-?(TGF-?).Research the relationship between the subtypes of HR-HPV and the concentration of these cytokines.Detect the single nucleotide polymorphism(SNP)of IL-6 and look for its effect upon the persistence of HR-HPV infection and the level of IL-6.All these works are aimed at providing theoretical foundation for the prevention of HR-HPV infection and cervical lesions progression.Methods:A total of 210 married women who were referred to colposcopy outpatients of Tianjin Central Hospital of Gynecology and Obstetrics from November 2015 to June 2016 were involved in our research.All subjects signed informed consent.According to the HR-HPV infection status,patients were divided into two groups: the cleared group(n=105)and the persistent group(n=105).According to the pathological results the persistent group were further divided into three groups: non squamous intraepithelial lesion(NSIL)35 cases?low grade squamous intraepithelial lesion(LSIL)35 cases and high grade squamous intraepithelial lesion(HSIL)35 cases.Our detection programs covered vaginal lavage fluid and Thinprep Cytologic Test.Flow cytometry BD CBA(cytometric bead array)and Elisa(enzyme-linked immunosorbent assay)were performed to detect 8 cytokines in the vaginal lavage fluid.The DNA was extracted from cervical exfoliated cells and used for genotyping of HR-HPV and detection of the SNP of IL-6 rs1800795(-174 G/C)?rs1800796(-572 C/G)by nest PCR(polymerase chain reaction)and pyrosequencing.SPSS 17.0 was used for data analysis.The quantitative data was expressed by means±standard.One-way ANOVA was used for the comparison among groups and LSD-t was further applied to compare the difference between two groups.If we need to compare the difference only between two groups,t test is the main method.Chi-square test was used for count data and a p value less than 0.05 was considered of great difference.Results:1.Compared with the cleared group,the concentration of IL-2 was lower in the persistent group and the level significantly decreased with the severity;while the level of IL-6 increased with the severity.2.Their was no relationship between the persistence of HR-HPV and the status of menopausal.3.The most common subtype of HR-HPV was HPV16(50.48%).4.Compared with other high risk genotypes,no difference was found about cytokines levels caused by HPV 16.5.Their was no difference between the two groups about the IL-6 rs1800795(-174G/C)?rs1800796(-572 C/G)frequencies of genotype and allele.6.The levels of IL-6 were not related with the SNP of rs1800795(-174G/C)?rs1800796(-572 C/G).Conclusions:1.The detection of cytokines in the vaginal lavage fluid by BD CBA provides a new way to study the changes of local immune microenvironment in the process of HR-HPV persistent infection.2.IL-2 and IL-6 play an essential role in the process of HR-HPV persistent infection.Their is a drift from Th1 to Th2 in the process of HR-HPV persistent infection.3..The most common subtype of HR-HPV was HPV16,but compared with other high risk genotypes,no difference was found about cytokines levels caused by HPV 16.4.The study of IL-6 SNP provides a new way to analyze the causes of HR-HPV persistent infection,founding the polymorphism associated with HR-HPV infection will provide a new theoretical basis for the prevention of HR-HPV infection and the progression of disease.