Vaginal Microbiota Changes Caused by HPV Infection in Chinese Women

Abstract

Human papillomavirus (HPV) infection is one of the most common sexually transmitted diseases. After studying 602 unvaccinated Chinese women using 16S rRNA to detect cervical-vaginal microecology, we analyzed the relationship between HPV infection and vaginal microecology including 20 HPV types. In Chinese women, L. gasseri-dominated and L. jensenii-dominated clusters were significantly absence. Microbial alpha diversity was significantly higher in HPV-infected and cervical intraepithelial neoplasia (CIN)-diagnosed groups than in healthy control group. Certain bacteria were associated with HPV infection and CIN, including Streptococcus, Prevotella, Chlamydia, Bifidobacterium, Ralstonia, and Aerococcus. With the development of disease, the proportions of community state type III (CST-III) and CST-IV-B gradually increased, whereas the proportions of CST-I and CST-IV-A gradually decreased. In addition, age was an influential factor for HPV infection. With aging, the probability of HPV infection and the proportion of CST-IV-B increase. In conclusion, our study was a large cross-sectional study that evaluated the relationship between vaginal microbiota and HPV infection, and brought essential comparable data.

Keywords: HPV infection; L. gasseri–dominated; age; cervical intraepithelial neoplasia; community state types (CST); vaginal microbiota.

Figure 1

Samples were classified by HPV subtypes. (A) The percentage of HC group, simple-infection group, and multiple-infection group in all samples. (B) The percentage of special HPV types in all samples. (C) HPV prevalence in the HPV-infected and CIN-diagnosed groups. The top HPVs are HPV16, 52, 58; 2HR-HPV; and HR-HPV + LR-HPV.

Figure 2

Histogram of relative abundance of species. (A) Vaginal microbiota at the species level in HC, HPV, and CIN groups. (B) Vaginal microbiota at the species level in HC, HPV-S, HPV-M, CIN1, CIN2, and CIN3 groups. (C) Vaginal microbiota at the species level from healthy control group women. (D) Vaginal microbiota at the species level from HPV-infected group women. (E) Vaginal microbiota at the species level from CIN group women.

Figure 3

Microbial alpha diversity analysis based on Shannon index. (A) Microbiota diversity (Shannon) comparison among groups of healthy control women, HPV-infected women and CIN-diagnosed women (Wilcoxon one-side test; ***p < 0.0001). (B) Microbiota diversity comparison of healthy control group, and groups infected with single and multiple HPV types (Wilcoxon one-side test; ***p < 0.0001). (C) Microbiota diversity comparison of healthy control group, and groups infected with high-risk (HR) and low-risk (LR) HPV types (Wilcoxon one-side test; **p < 0.005 and ***p < 0.0001). (D) Microbiota diversity comparison of healthy control group, and groups diagnosed with CIN1, CIN2, and CIN3 (Wilcoxon one-side test; **p < 0.005 and ***p < 0.0001). (E) Microbiota diversity among participants infected with different HPV types in comparison with healthy women (Wilcoxon one-side test; *p < 0.05 and **p < 0.005).

Figure 4

Bacterial Taxa Analysis. (A) Streptococcus was significantly higher in HPV-infected group than HC group. (B, C) Prevotella and Chlamydia were the bacterial species that significantly higher in HPV- infected group and CIN-diagnosed group than HC group. (D, E) Bifidobacterium and Ralstonia were significantly lower in HPV-infected group and CIN-diagnosed group than HC group. (F) Aerococcus was lower in HPV-infected women and higher in CIN-diagnosed women compared with HC women. (G) Cladogram of the LEfSe analysis of the vaginal microbiota in the three study groups. The microbial compositions were compared at different evolutionary levels. (H) LDA scores obtained from the LEfSe analysis of the vaginal microbiota in the different groups. An LDA effect size of >2 was used as a threshold for the LEfSe analysis. LDA, linear discriminant analysis; LEfSe, LDA effect size analysis. Wilcoxon one-side test; *P < 0.05 and **P < 0.005. HC, healthy control group; HPV, HPV-infected group; CIN, CIN-diagnosed group.

Figure 5

Samples were classified by CST and principal coordinates analysis (PCoA) of microbiota data based on Bray–Curtis distance of vaginal microbiota. (A–H) According to community state types (CSTs), the VMB of women was divided into I-V subtypes (n, the number of samples; HC, healthy control group; HPV, HPV-infected group; CIN, CIN-diagnosed group; HPV-S, HPV simple infection group; HPV-M, HPV multiple infection group; CIN1, CIN1 group; CIN2, CIN2 group; CIN3, CIN3 group). (I) Principal coordinates analysis (PCoA) of microbioal species data based on Bray–Curtis distance matric demonstrated six main vaginal microbiota clusters.

Figure 6

HPV infection and CST subtypes based on age. (A) Different CST subtypes in all samples according to age. (B) HPV prevalence and CIN-diagnosed in all samples according to age.