Influence of human papillomavirus type 16 (HPV-16) E2 polymorphism on quantification of HPV-16 episomal and integrated DNA in cervicovaginal lavages from women with cervical intraepithelial neoplasia

Abstract

Integrated human papillomavirus type 16 (HPV-16) viral loads are currently estimated by quantification with real-time PCR of and E2 (RT-E2-1) DNA. We assessed the influence of HPV-16 E2 polymorphism on quantification of integrated HPV-16 DNA in anogenital specimens. HPV-16 E2 was sequenced from 135 isolates (123 from European and 12 from non-European lineages). An assay targeting conserved HPV-16 E2 sequences (RT-E2-2) was optimized and applied with RT-E6 and RT-E2-1 on 139 HPV-16-positive cervicovaginal lavages collected from 74 women [58 human immunodeficiency virus (HIV)-seropositive and 16 HIV-seronegative]. Ratios of HPV-16 copies measured with RT-E2-2 and RT-E2-1 obtained with African 2 (median53.23, range51.92-3.49) or Asian-American (median53.78, range51.47-37) isolates were greater than those obtained with European isolates (median51.02, range50.64-1.80; P,0.02 for each comparison). The distribution of HPV-16 E2 copies measured in 139 samples with RT-E2-2 (median56150) and RT-E2-1 (median58960) were different (P,0.0001). The risk of high-grade cervical intraepithelial neoplasia (CIN-2,3) compared with women without CIN was increased with higher HPV-16 total [odds ratio (OR)52.17, 95 % confidence interval (CI)51.11-4.23], episomal (OR52.14, 95 % CI51.09-4.19), but not for , after controlling for age, race, CD4 count, HIV and HPV-16 polymorphism. The proportion of samples with an E6/E2 ratio .2 in women without squamous intraepithelial lesion (7 of 35) was similar to that of women with CIN-2,3 (5 of 11, P50.24) or CIN-1 (5 of 14, P50.50). HPV-16 E2 polymorphism was a significant factor that influenced measures of HPV-16 integrated viral load.

Sequence variation analysis of the E2 gene of human papilloma virus type 16 in cervical lesions from women in Greece

“…Of the 29 sequence variations, 19 resulted in amino acid changes in the three HPV16 E2 protein domains (Tables 2, 3). The majority of the nucleotide variations identified have also been reported in previous studies [40][41][42][43][44][45][46][47][48][49][50], but three new variations were reported for the first time in the present report. In the 5 0 region of the E2 gene, which encodes the transactivation domain of the protein, two new variations were detected at position 2791 and 2823 in cervical cases that were associated with HPV16 European variants (Tables 2, 3).…”

Human papillomavirus HPV-16, 18, 52 and 58 integration in cervical cells of HIV-1-infected women

“…These limitations could underestimate the detection of integrated forms. [11][12][13] However, our results are in agreement with previous studies, [14][15][16] suggesting that the viral integration of HPV could be a good biomarker for the evolution from HPV infection to cervical cancer.…”

Disruption of the E2 Gene Is a Common and Early Event in the Natural History of Cervical Human Papillomavirus Infection: A Longitudinal Cohort Study

“…When designing primers, the National Center for Biotechnology Information (NCBI) database was interrogated to target regions of homology across all registered HPV16 and HPV18 variants. We also reviewed reports identifying polymorphisms in the HPV16 and HPV18 E2 genes with a view to ensuring that disruption of E2 could not be explained by the failure of primer binding due to sequence variation (15)(16)(17)(18)(19)(20)(21)(22)(23).…”