Recombinant HPV16 L1 protein

The human leukocyte antigen (HLA) locus on chromosome 6 has been reported to be associated with cervical cancer. We investigated two independent single-nucleotide polymorphisms in a large case-control series of cervical dysplasia and carcinoma that has been newly established by the German Cervigen Consortium, comprising a total of 2481 cases and 1556 healthy females. We find significant associations for both variants, rs9272117 atHLA-DQA1and rs2844511 atMICAandHCP5, with cervical disease. Both variants showed evidence of association with invasive cervical cancer (rs9272117: OR 0.89, 95% CI 0.79-0.99,P= .036; rs2844511: OR 1.17, 95% CI 1.04-1.31,P= .008) and with high-grade dysplasia (rs9272117: OR 0.78, 95% CI 0.70-0.87,P= 7.1 x 10(-6); rs2844511: OR 1.13, 95% CI 1.01-1.26,P= .035), as well as in a combined analysis of both groups (rs9272117: OR 0.83, 95% CI 0.75-0.91,P= 6.9 x 10(-5); rs2844511: OR 1.14, 95% CI 1.04-1.26,P= .005). Variant rs2844511, but not rs9272117, also showed modest evidence of association with low-grade dysplasia (OR 1.26, 95% CI 1.04-1.54,P= .019). In case-only analyses, rs2844511 tended to predict HPV status (P= .044) and rs9272117 tended to associate with HPV16 (P= .022). RNA studies in cervical samples showed a significant correlation in the transcript levels ofMICA,HCP5andHLA-DQA1, suggesting extensive co-regulation. All three genes were upregulated in HPV16-positive samples. In stratified analyses, rs9272117 was associated withHLA-DQA1levels, specifically in HPV-positive samples, while rs2844511 was associated withMICAandHCP5levels. The risk allele of rs2844511 was required for correlations betweenMICAorHCP5withHLA-DQA1. Altogether, our results support 6p21.32-33 as the first consistent cervical cancer susceptibility locus and provide evidence for a link between genetic risk variants, HPV16 status and transcript levels ofHLA-DQA1,HCP5andMICA, which may contribute to tumor immune evasion.

The association of microbiome imbalance with cancer development is being one of the research hotspots. Persistent HPV infection is a causal event in cervical cancer initiation, but, little is known about the microbiome composition and function in HPV infection. Here we identified the compositional and functional alterations on vaginal samples from 27 HPV16 positive women and 25 age-matched HPV negative controls using shotgun metagenomic sequencing, to provide a comprehensive investigation describing the microbial abundances and enriched metabolic functions in cervicovaginal metagenomes. We further employed qPCR assays to evaluate two selected gene markers of HPV16 infection in an independent validation cohort consisting of 88 HPV16 positive women and 81 controls, and six selected species markers in a subset of validation cohort of 45 HPV16 positive women and 53 controls. We found that the relative abundance of dominant Firmicutes was lower, Actinobacteria, Fusobacteria and viruses phyla were significantly higher in the HPV16-positive group; 77 genera includingGardnerella, Peptostreptococcus, andPrevotellawere higher, and 20 genera includingLactobacillusandAerococcuswere lower in the HPV16-positive women. Abundance of 12 genes, 17 genera, and 7 species biomarkers showed an excellent predictive power for the HPV16-positive individuals, with 0.861, 0.819, and 0.918, respectively, of the area under the receiver-operating characteristic curve (AUC). We further characterized the microbial function, and revealed that HPV16-positive women were enriched in metabolism and membrane transport, and depleted by glycan biosynthesis and metabolism, and replication and repair. Quantitative PCR measurements validated that one gene marker and three species were significantly enriched in HPV16-positive women. These results highlight a fundamental fact that there are altered composition and function of the vaginal microbiome in HPV16-positive women, suggesting that vaginal dysbiosis may be associated with HPV infection in the female genital tract.