Kawahara Rina, Fujii Takuma, Kukimoto Iwao, Nomura Hiroyuki, Kawasaki Rie, Nishio Eiji, Ichikawa Ryoko, Tsukamoto Tetsuya, Iwata Aya
Department of Obstetrics and Gynecology, Fujita Health University, School of Medicine, 1-98, Dengakugakubo, Toyoake, Aichi, 470-1192, Japan.
Pathogen Genomics Center, National Institute of Infectious Diseases, 4-7-1, Gakuen, Musashi-murayama, Tokyo, 208-0011, Japan.
Sci Rep. 2021 Jan 25;11(1):2156. doi: 10.1038/s41598-020-80176-6.
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.
与免疫调节相关的持续性人乳头瘤病毒（HPV）感染可能导致高级别鳞状上皮内病变（CIN）2/3。目前，关于宫颈阴道微生物群、局部细胞因子水平以及与CIN相关的HPV感染的信息较少。对局部手术后患者的随访为监测宫颈阴道环境的变化提供了机会。因此，我们进行了这项纵向回顾性研究，以确定41名日本CIN患者中HPV基因型、宫颈阴道微生物群和局部细胞因子谱之间的关联。通过对基因组DNA进行PCR，使用针对V3/4区域的通用16S rRNA基因（rDNA）细菌引物来鉴定宫颈阴道微生物群，随后进行MiSeq测序。我们发现手术后阴道阿托波菌显著减少（p < 0.047），而解脲脲原体增加（p < 0.022）。通过基于多重微珠的免疫测定法测量宫颈黏液中的细胞因子水平，结果显示手术后白细胞介素-1β（IL-1β，p < 0.006）、肿瘤坏死因子-α（TNF-α，p < 0.004）、巨噬细胞炎性蛋白-1α（MIP-1α，p < 0.045）和嗜酸性粒细胞趋化因子（p < 0.003）显著降低。值得注意的是，手术后嗜酸性粒细胞趋化因子水平与HPV清除平行下降（p < 0.028）。因此，局部手术影响了宫颈阴道微生物群、HPV感染状态和免疫反应。宫颈上皮内瘤变手术后宫颈阴道微生物群和宫颈细胞因子谱的变化可能对未来理解CIN的发病机制很重要。
