# A prospective cohort study on the association between cervical microenvironmental factors and the efficacy of treating high-risk human papillomavirus infection comorbid with cervical diseases

## TL;DR

This study explores how the cervical environment affects the success of interferon-based treatment for high-risk HPV infections and cervical abnormalities.

## Contribution

The study identifies specific microenvironmental and immune factors associated with successful HR-HPV clearance following interferon therapy.

## Key findings

- The clearance group showed higher baseline expression of TRAF3IP3, ZBP1, and IFI35, and increased immune activation pathways.
- Treatment failure was linked to reduced Lactobacillus and increased Gardnerella and other bacterial species in the cervical microbiome.
- Cervical secretory cytokines like IL-2, IL-8, and IL-12p70 showed differences between groups, while IL-4 and IL-5 were undetectable.

## Abstract

Interferon-based local therapy is an intervention for high-risk human papillomavirus (HR-HPV)-associated low-grade squamous intraepithelial lesions (LSIL) or lower-grade cervical abnormalities. This study sought to delineate the differences in clinical outcomes following interferon-based local drug treatment and elucidate the microenvironmental factors driving these disparities.

Cervical secretions, cell brush specimens, and cervical tissue samples were collected from patients with persistent HR-HPV infection and LSIL/lower-grade lesions at Shanghai First Maternity and Infant Hospital. Follow-up samples were obtained at 3 months post-treatment. Cervical secretions were subjected to 16S rRNA sequencing (to profile the microbiota) and cytokine quantification. Cell brush specimens were analyzed via transcriptome sequencing, while cervical tissue samples underwent immunohistochemical staining. Efficacy-related markers were assessed through both inter-group (independent comparisons) and intra-patient (self-paired) analyses.

At the transcriptome level, the HR-HPV clearance group exhibited lower enrichment in pathways related to differentiation, keratinization, and development but higher enrichment in immune activation pathways compared to the persistence group at baseline (with a reversed pattern observed at follow-up). Baseline expression of TRAF3IP3, ZBP1, and IFI35 was higher in the clearance group, and ZDHHC11 expression remained consistently elevated. Immunohistochemical findings further demonstrated that the percentage of TRAF3IP3- and ZBP1-positive cells at baseline was significantly higher in the clearance group than in the persistence group. At the microbial level, treatment failure was associated with reduced Lactobacillus abundance, increased Gardnerella, Streptococcus anginosus, Schaalia turicensis, and Comamonadaceae abundance, alongside higher alpha diversity. Among cervical secretory cytokines, IL-2, IL-8, IL-12p70 showed inter-group differences, while IL-4 and IL-5 were barely detectable.

This study characterizes the cervical microenvironmental differences underlying divergent responses to interferon-based therapy, highlighting that coordinated changes in the microenvironment and immune status modulate treatment outcomes. The upregulated mRNA and protein levels of TRAF3IP3 and ZBP1 in the baseline period favor HR-HPV clearance, suggesting their potential as promising therapeutic targets.