Changes to the cervicovaginal microbiota and cervical cytokine profile following surgery for cervical intraepithelial neoplasia

Rina Kawahara,Eiji Nishio,Takuma Fujii,Aya Iwata,Tetsuya Tsukamoto,Rie Kawasaki,Ryoko Ichikawa,Hiroyuki Nomura,Iwao Kukimoto

doi:10.1038/s41598-020-80176-6

Abstract

Persistent HPV infection associated with immune modulation may result in high-grade squamous intraepithelial lesions (CIN)2/3. Currently, there is little information on the cervicovaginal microbiome, local cytokine levels and HPV infection related to CIN. Follow-up of patients after local surgery provides an opportunity to monitor changes in the cervicovaginal environment. Accordingly, we undertook this longitudinal retrospective study to determine associations between HPV genotypes, cervicovaginal microbiome and local cytokine profiles in 41 Japanese patients with CIN. Cervicovaginal microbiota were identified using universal 16S rRNA gene (rDNA) bacterial primers for the V3/4 region by PCR of genomic DNA, followed by MiSeq sequencing. We found that Atopobium vaginae was significantly decreased (p < 0.047), whereas A. ureaplasma (p < 0.022) increased after surgery. Cytokine levels in cervical mucus were measured by multiplexed bead-based immunoassays, revealing that IL-1β (p < 0.006), TNF-α (p < 0.004), MIP-1α (p < 0.045) and eotaxin (p < 0.003) were significantly decreased after surgery. Notably, the level of eotaxin decreased in parallel with HPV clearance after surgery (p < 0.028). Thus, local surgery affected the cervicovaginal microbiome, status of HPV infection and immune response. Changes to the cervicovaginal microbiota and cervical cytokine profile following surgery for cervical intraepithelial neoplasia may be important for understanding the pathogenesis of CIN in future.

Highlights

Persistent human papillomaviruses (HPVs) infection associated with immune modulation may result in high-grade squamous intraepithelial lesions (CIN)2/3
Transient HPV infections cause low-grade squamous intraepithelial lesions (CIN1), whereas persistent HPV infection presumably associated with accumulating genetic changes and immune modulation can result in highgrade squamous intraepithelial lesions (CIN2–3) or cervical cancer
We compared the cervicovaginal microbiota in women with cervical intraepithelial neoplasia (CIN) before and after surgery to assess whether there were any correlations between bacterial composition and status of HPV infection (Figure S1)

Summary

Introduction

Persistent HPV infection associated with immune modulation may result in high-grade squamous intraepithelial lesions (CIN)2/3. Follow-up of patients after local surgery provides an opportunity to monitor changes in the cervicovaginal environment We undertook this longitudinal retrospective study to determine associations between HPV genotypes, cervicovaginal microbiome and local cytokine profiles in 41 Japanese patients with CIN. It was difficult to compare the relationships between HPV infections, microbiota and local host immune response in the cohort of patients as a whole, but monitoring the same individual patients with CIN over time after surgery avoid this problem of heterogeneity. Surgery may change the microbial diversity[4] and the cervicovaginal environment, including immune r esponses[5] and the status of HPV infection[6] We undertook this retrospective longitudinal study to elucidate associations between cervicovaginal microbiota, HPV infection and cytokine profiles in individual premenopausal women with CIN before and after surgery

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