Relationship Between Vaginal Immune Microenvironment Before Treatment And Postoperative HPV Outcome In Patients With HSIL

BackgroundCervical cancer is the fourth most malignant tumor in women and one of the leading causes of death in women.HPV is a very common virus transmitted through sexual contact.Almost all cases of cervical cancer（99%）are related to high-risk human papillomavirus（HPV）infection.Cervical intraepithelial neoplasia is characterized by cell changes in the cervical transformation area.Cin2 / 3 lesions,namely high-grade squamous intraepithelial lesions（HSIL）,are related factors of cervical precancerous lesions.At present,resection is the first choice for the treatment of HSIL patients,including circular electrotomy,cold knife conization and laser conization biopsy.However,long-term follow-up data have showed that compared with the general population,women treated with CIN still have an increased risk of invasive cervical cancer for at least 10 years and possibly up to 20 years after treatment.HR-HPV persistent infection after conization is an independent risk factor for the recurrence of HSIL.The predictive ability of postoperative HPV based detection for persistent / recurrent histological HSIL was 91%.It can be seen that the outcome of HPV after conization has a very important impact on the prognosis of patients.The female lower genital tract is composed of cervix,vagina and vagina.The female lower genital tract is composed of a variety of immune cells,accounting for 6%to 20% of all cells,which can prevent the invasion of pathogens.HPV can effectively escape the immune response of the body,so as to establish the continuous infection of the virus.The continuous HPV infection may be related to the cumulative genetic changes and immune regulation.The role of inflammation in HPV infection and disease is multifaceted: on the one hand,clearing HPV infection involves cytotoxic immune response;On the other hand,tumor related inflammation may promote carcinogenesis by secreting growth promoting cytokines,chemokines,angiogenic factors and proteases by inflammatory cells.The inflammatory environment in tumors can also actively inhibit specific antitumor responses.Innate immune cells play a key role in tumor clearance.Tumor associated macrophages（TAMs）are macrophages involved in the formation of tumor microenvironment,which can play different antitumor or antitumor roles.According to their functional differences,TAMs can be roughly divided into two subgroup types,namely M1 type and M2 type.M1 macrophages produce proinflammatory cytokines such as IL-6,and TNF-α;M2 macrophages have anti-inflammatory and immunomodulatory effects and produce anti-inflammatory cytokines such as IL-10 and TGF-β.IL-6 and TNF-α are two pleiotropic cytokines that play a role in maintaining the balance of the immune system,inflammation and tumor formation.It can be seen that female vaginal immune microenvironment is closely related to cervical lesions,which affects the prognosis and outcome of the disease to a certain extent.However,there are few studies on the correlation between preoperative vaginal immunity microenvironment of HSIL and postoperative HPV infection.ObjectiveTo compare the correlation between the concentration of IL-6 and TNF-α in vaginal lavage fluid before treatment and postoperative HPV outcome in patients with HSIL,analyze the changes of preoperative cervical immune microenvironment in patients with different HPV outcomes,and select the proper cytokines that can be used to predict the postoperative HPV outcome of patients.MethodIn this study,we selected and tested the previously collected samples from the patients who underwent surgical treatment for HSIL in the second hospital of Jilin University from December 2018 to October 2019.The general clinical data of the patients were collected and the vaginal lavage fluid was taken.Patients were grouped according to the results of HPV reexamination 3-4 months after operation.According to the inclusion and exclusion criteria,the total number of patients finally included was 64,including 10 cases in the postoperative HR-HPV positive group and 36 cases in the postoperative HR-HPV negative group.At the same time,according to the exclusion criteria,18 patients with HPV and TCT screening and negative results in the second hospital of Jilin University were selected as the healthy control group.Enzyme linked immunosorbent assay（double antibody sandwich method）was used to determine the concentrations of representative cytokines IL-6 and TNF-α of M1 macrophages in lavage solution.Appropriate statistical methods were selected to compare the differences between the concentrations of IL-6 and TNF-α in different groups,analyze the correlation between them and postoperative HPV outcome,and explore their predictive value for postoperative HPV outcome.Result1.There was no significant difference in the general clinical data（age,preoperative HR-HPV type,TCT results,operation method and pathological type）among the healthy control group,postoperative HPV negative group and postoperative HPV positive group.2.The concentration of IL-6 in preoperative vaginal lavage fluid increased significantly in postoperative HPV negative group,but there was no significant difference between postoperative HPV positive group and control group;The concentration of TNF-α decreased significantly in the postoperative HPV positive group,but there was no significant difference between the postoperative HPV negative group and the control group.3、The decrease of TNF-α（P = 0.006,P < 0.01）is an independent risk factor for postoperative HR-HPV positive.For each unit of TNF-α decrease,the risk of HR-HPV infection in patients with HSIL after conization increased by 1.03 times.4.IL-6 and TNF-α both have predictive value for the prognosis after HSIL conization（AUC is 0.878 and 0.914 respectively）.In preoperative vaginal lavage fluid of HSIL patients,if the concentration of IL-6 is less than 6.10ng/l and the concentration of TNF-α is less than 155.31ng/l,patients are more likely to be positive for HR-HPV after HSIL conization.Conclusion1.Age,surgical procedure,preoperative HR-HPV type,TCT results and pathological grade had no correlation with postoperative HPV outcome;2.The polarization state of macrophages may affect the clearance of HPV in patients with HSIL;3.IL-6 and TNF-α are associated with HPV outcome after treatment of HSIL.TNF-α is an independent protective factor for HPV outcome after operation.The smaller the concentration of TNF-α in the preoperative immune microenvironment,the higher the risk of HR-HPV positive after HSIL conization;4.Compared with IL-6,TNF-α has more predictive value for the prognosis after HSIL conization.If the concentration of IL-6 is less than 6.10ng/l and the concentration of TNF-α is less than 155.31ng/l in preoperative vaginal lavage fluid of HSIL patients,they are more likely to be positive for HR-HPV after HSIL conization.And these patients may require closer postoperative follow-up.