Vaginal microbiome composition was demonstrated to be associated with cervical disease. The colonization characteristics
of vaginal microbes and their association with the different cervical disease status, especially cervical cancer (CC), are
rarely investigated. In this cross-sectional study, we characterized the vaginal microbiome of women with different status of
cervical diseases, including 22 NV + (normal tissue with HPV infection), low-grade squamous intraepithelial lesion (LSIL,
n = 45), high-grade squamous intraepithelial lesion (HSIL, n = 36) and CC (n = 27) using bacterial 16S DNA sequencing.
Thirty HPV-negative women with normal tissue were used as the control group. We found that higher diversity of microbiome
with gradual depletion of Lactobacillus, especially L. crispatus, was associated with the severity of cervical disease.
High-risk HPV16 infection was associated with higher microbiome diversity and depletion of Lactobacillus in high-grade
cervical diseases (i.e. HSIL and CC). The CC group was characterized by higher levels of Fannyhessea vaginae, Prevotella,
Bacteroides, Finegoldia, Vibrio, Veillonella, Peptostreptococcus, and Dialister. Co-occurrence network analyses showed that
negative correlations were exclusively observed between Lactobacillus and other bacteria, and almost all non-Lactobacillus
bacteria were positively correlated with each other. In particular, the most diverse and complex co-occurrence network of
vaginal bacteria, as well as a complete loss of L. crispatus, was observed in women with CC. Logistic regression model
identified HPV16 and Lactobacillus as significant risk and protective factors for CC, respectively. These results suggest that
specific Lactobacillus species (e.g. L. crispatus and L. iners) can be used as important markers to target prevention measures
prioritizing HPV16-infected women and other hrHPV-infected women for test, vaccination and treat initiatives.