The Association Between HR-HPV Infection,cervical Cancer And Vaginal Microecology Based On Natural Population

ObjectiveTo investigate the status of HPV infection,vaginitis,vaginosis,vaginal microbiota of Chinese women,and the association of BV,VVC,TV,HR-HPV infection with abnormal cervical cytology in the participants.To demonstrate the vaginal microbiota associated with BV,HPV infection(including multiple infections and different HPV types)and CIN,as well as the vaginal microbiota associated with HPV persistent infection and regression.Materials and MethodWomen were recruited from cervical cancer screening site,Wuxiang,Shanxi Province and Yanting,Sichuan Province in 2018-2019.Cervical exfoliated cell specimens and reproductive tract secretions samples were collected.BV,VVC and TV were diagnosis by real-time Polymerase Chain Reaction(PCR)and HPV genotype was determined by Multiplex PCR and reverse hybridization.Microbial composition was determined by 16S rRNA sequencing.The Logistic multivariate model was used to analyze the risk factors of abnormal cervical cytology.QIIME2 and R were used to analyze the 16S rRNA gene sequence data to compare the differences in the diversity and relative abundance of the vaginal microbiota among different groups.Results1.A total of 645 women including 408 NILM,182 ASC-US,23 LSIL,and 32 HSIL were included in the study.The positive rates of HR-HPV(14.7%,39.0%,78.3%and 87.5%)increased with the severity of cytology(Ptrend<0.001).The prevalence of BV in different cytological diagnosis(34.1%,49.5%,60.9%and 43.8%)were significantly different(Ptrend=0.001),and the prevalence of BV in ASC-US and LSIL were significantly higher than that in women with NILM(P=0.003;P=0.05).The prevalence of VVC in different cytological diagnosis(14.2%,4.9%,8.7%and 3.1%)was significantly different(P=0.004),and the ASC-US group was significantly lower than the NILM group(P=0.004).There was also a significant difference in the distribution of the prevalence of TV(2.5%,9.3%,4.3%,and 6.3%,P=0.003).The prevalence of TV in women with abnormal cytology was higher than that in women with normal cytology(P=0.002;P=0.038).The Logistic analysis showed that HR-HPV infection,BV,TV,VVC,lower level of education,and postmenopause were related to abnormal cervical cytology.Lower level of education,postmenopause,BV,TV and HPV infection were independent risk factors for abnormal cervical cytology;VVC was a protective factor for abnormal cervical cytology(P<0.05).2.With the increase of histological grade,the positive rate of HPV significantly increased(Ptrend<0.001).However,there was no significant difference in the prevalence of BV,VVC and TV among the groups(P>0.05).3.The composition of vaginal microbiota of HPV positive and negative groups was significantly different.The alpha and beta diversities of HPV positive women were significantly higher than that of HPV negative women.The relative abundance of a total of 27 genera is different between HPV positive and negative groups.The set of 27 bacterial genera was identified to differentiate HPV+from HPV-(AUC=0.652).Lactobacillus was selected as the most influential predictor in this model,following by Gardnerella and Prevotella.4.From the non-BV&HPV-group(414 HPV-negative women without BV)to the non-BV&HPV+group(108 HPV-positive women without BV),to the BV&HPVgroup(330 HPV-negative women with BV)and then to the BV&HPV+group(163 HPV positive-women with BV),microbial diversity increased.The relative abundance of 13 genera,including Gardnerella,Prevotell,and Sneathia increased,whilst Lactobacillus declined.5.Multiple HPV infection,certain HPV genotype infection and cervical intraepithelial neoplasia(CIN)status were associated with some HPV&BV-associated microbiota and higher microbial diversity.6.Compared with HPV-negative women,women with new HPV infections have a higher microbial diversity and different microbial composition.Conclusion1.Except HPV infection,BV,TV,and VVC are all important influencing factors of abnormal cervical cytology,but they are mainly related to ASC-US and LSIL.In addition,lower level of education and postmenopause may also increase the possibility of abnormal cervical cytology.2.Vaginal microbiota was associated with BV,HPV infection(including multiple HPV infections and certain specific HPV genotypes)and CIN.BV,HPV infection,multiple HPV infection and CIN2+increase the diversity of microbial flora.3.Lactobacillus is a protective factor for HPV infection and cervical cancer;Prevotella and Gardnerella are significantly related to BV and HPV infection,and they are the most impactful predictors for predicting HPV infection.The association between certain genera and HPV infection were controversial due to BV.4.HPV&BV-assocaited vaginal microbiota and HPV infection are correlated to low-grade cervical lesions.In high-grade lesions,HPV is the main influencing factor.The association between vaginal microbiota and high-grade cervical lesions is disputable.