Relationship Between Vaginal Microbiota And Human Papillomavirus 16 Infection Among Women With Colposcopy In Gynecology Clinic

Objective:Human papillomavirus(HPV)is one of the most common sexually transmitted viruses.Persistent infection of high-risk HPV(HR-HPV)infection is closely associated with the occurrence of various genital cancers such as cervical cancer,vaginal cancer and vulvar cancer.Vaginal microbiota is closely related to pathogen infection and female reproductive tract health.It is essential to maintain the balance and stability of vaginal micro-ecology.HPV16 is the most common carcinogenic type of HPV,which will change the structure of vaginal microbiota during the process of infecting female reproductive tract.However,the relationship between vaginal microbiota and HPV16 infection is still unclear.This study was conducted on women with HPV infection or abnormal cervical cytology.Based on the analysis of the characteristics of HPV16 infection and vaginal microbiota,the relationship between the changes of vaginal microbiota and HPV16 infection was explored.Further predicted the functions of vaginal microbial using PICRUSt(Phylogenetic Investigation of Communities by Reconstruction of Unobserved States)software.This study aimed to develop new ideas for the development of strategies to prevent and control HPV16 infection-related diseases,and provide new clues for the mechanism research of HPV16 infection-related diseases.Methods:A total of 1396 women who underwent colposcopy in the Department of Gynecology,Second Hospital of Shanxi Medical University between April 2017 and June 2018 were enrolled in this study.A structured questionnaire was used to collect the related characteristics concerning HPV infection,and cervical exfoliated cells were collected.The HPV infection status and its typing were detected by flow-through hybridizationtechnology.After excluded women who used antibiotics within 2 weeks before enrollment further and HPV16 multiple infections,68 study participants enrolled from May to June,2018 were selected for 16 S rDNA sequencing.Sequences were clustered into OTU(operational taxonomic unit)at the similarity of 97% by Usearch.The taxonomy compositions and abundances were calculated by Na?ve Bayesian assignment.LEfSe(linear discriminant analysis effect size)was used to detect the characteristic microbiota among different groups.Microbial functions were predicted using PICRUSt software.All the data analyse were performed using SPSS22.0.Graphics were generated using R3.4.3and GraphPad Prism 7.Results:1.HPV infection characteristics: HPV infection was dominated by single infection(73.2%),which was dominated by single HR-HPV infection(72.0%).HPV16(38.6%),52(15.2%),58(15.1%),53(10.5%),18(8.7%)were the most common genotypes.The infection rates of HPV and HR-HPV in women aged ?35 years old were higher than women 46~55 years old,>55 years old.The infection rates of HPV16 and HPV16 single infection in women with an education level of college degree or above were lower than that in junior high school,primary school and below.The infection rates of HPV16 and HPV16 single infection in unmarried,divorced and populations in other marital status were higher than those in married people.The infection rates of HPV in smokers were higher than others.The infection rates of HPV,HR-HPV,HPV16 and HPV16 single infection in passive smokers were higher than others.2.Vaginal microbiota characteristics: A total of 8711 OTUs,31 vaginal phylum,and466 vaginal genus were obtained.The main vaginal phylum were Firmicute,Actinobacteria,Bacteroidetes,Fusobacteria and Tenericutes,accounting for 75.4%,15.7%,4.4%,2.5% and1.4%,respectively.The main vaginal genus were Lactobacillus,Gardnerella,Peptoniphilus,Atopobium and Sneathia,accounting for 62.4%,11.5%,3.8%,3.0% and 2.5%,respectively.3.Relationship between vaginal microbiota and HPV16 infection: Alpha diversity ofHPV16 positive group was higher than that in negative group.The abundance of Fusobacteria,Tenericutes,Peptoniphilus,Prevotella,Saccharofermentans,Sneathia,Streptococcus,Veillonella,Finegoldia,Ureaplasma,Bulleidia and Bifidobacterium in HPV16 infection group was higher than that in HPV16 negative group.The characteristic microbiota of HPV16 infection group was ureaplasma.The characteristic microbiota of HPV16 negative group were Peptostreptococcaceae,Actinomyces,Granulicatella,Burkholderiales,Betaproteobacteria.4.Functions prediction of vaginal microbiota: The functions of vaginal microbiota were focused on level of Metabolism,Genetic Information Processing,Environmental Information Processing,Cellular Processes,Human Diseases,Organismal Systems.At KEGG level 2,vaginal microbiota played an important role in Membrane Transport,Replication and Repair,Carbohydrate Metabolism,Translation,Amino Acid Metabolism,Nucleotide Metabolism and Energy Metabolism in HPV16 infection group and negative group.At KEGG level 3,the abundance of Apoptosis,Cyanoamino acid metabolism,Fatty acid biosynthesis,Lipid biosynthesis proteins,Phenylpropanoid biosynthesis,Propanoate metabolism,Tetracycline biosynthesis in HPV16 infection group were higher than that in HPV16 negative group.Conclusions:1.HPV16,52,58 were common infection types.In particular,it was found that HPV53 infection rate was high in this population.It is conducive to the prevention and control of HPV infection in this region,and can provide basis for further research of vaccines covering major types.HPV infection rate is higher in people aged ?35 years,smoking and passive smoking.HR-HPV infection rate is higher in people aged ?35 years and passive smoking.HPV16 and HPV16 single infection rates are higher in people junior high school and below,unmarried and divorced and passive smoking.It can provide a theoretical basis for the screening of HPV and HPV 16 infection.2.In HPV16 infection group and negative group,the main microbiota phylum wasFirmicutes,the main microbiota genus was Lactobacillus.Alpha diversity of HPV16 positive group was higher than that in negative group.In addition to the reported HPV-related microbiota,this study found that Peptoniphilus,et al may be associated with the HPV16 infection.It provided new ideas from the perspective of maintaining the balance of vaginal microbiota for the development of strategies to prevent and control HPV16 infection-related diseases.3.The function of vaginal microbiota was focused on Metabolism.HPV16 infection may be associated with the abundance of Apoptosis,et al.It is suggested that the vaginal microbiota may be involved in the pathogenesis of HPV16.This can provide new clues for the mechanism research of HPV16 infection.