Genetic polymorphism in human papillomavirus (HPV)-33 and -35 was investigated in 1055 sexually active women (732 human immunodeficiency virus [HIV] seropositive and 323 HIV seronegative).Consecutive genital specimens obtained at 6-month intervals were screened for HPV-33 and -35 by use of MY09-MY11. HPV-33 and -35 isolates from 95 women were analyzed by polymerase chain reaction sequencing of the long control region (LCR), E6, and E7.For HPV-33, 101 (20%) of 506 nucleotides in the LCR were variable, compared with 10 (2.1%) of 483 nucleotides in E6 (P<.001) and 6 (1.9%) of 324 nucleotides in E7 (P<.001). For HPV-35, the proportion of variable nucleotide sites was similar between the LCR and both E6 (P=.54) and E7 (P=.33). The presence of a 78-base pair deletion in HPV-33 (relative risk [RR], 1.8 [95% confidence interval [CI], 1.2-2.7]) and the presence of nonsynonymous E7 variations in HPV-35 (RR, 2.6 [95% CI, 1.4-4.6]) were associated with persistence. When the data for HPV-33 and -35 were combined, infection by HPV isolates with nonsynonymous E7 variations (RR, 2.3 [95% CI, 1.6-3.4]; P=.001) and ethnicity (P=.04) were associated with persistence, whereas age (P = .14) and HIV infection/CD4 cell count status (P=.12) were not significantly associated with persistence, by logistic regression analysis.HPV-33 and -35 polymorphism was different between types and was associated with persistence of HPV infection.
ABSTRACT Human papillomavirus type 16 (HPV-16) viral load in cervicovaginal lavage samples collected from 66 human immunodeficiency virus-seropositive women was inversely correlated with blood CD4 count ( P = 0.002). HPV-16 viral load was 81-fold higher in women with cervical smears suggestive of high-grade lesions (median, 4,425,883 copies/μg of DNA) than in women with normal smears (median, 54,576), controlling for age ( P = 0.006).
Human papillomavirus (HPV) type 52 DNA was detected in cervicovaginal lavage samples from 91 (12.4%) of 732 human immunodeficiency virus (HIV)–seropositive women and 23 (7.1%) of 323 HIV-seronegative women (P=.0004). HIV infection was an independent predictor for HPV-52 infection when controlling for age and sexual activity (odds ratio, 2.21; 95% confidence interval, 1.30–3.75: P=.003). We describe the genomic polymorphism of 114 HPV-52 isolates. Long control region (LCR) mutations defined 27 HPV-52 variants. Nearly 32% of HPV-52 isolates carried deletions in the LCR. E6 and E7 mutations defined 17 and 9 variants, respectively. Five nonsynonymous E6 mutations were clustered from amino acids 92 to 94, near the putative p53 binding area. White women were more frequently infected by the prototype strain than were women of African descent (P=.0001). The genetic diversity of HPV-52 should facilitate the investigation of the role of genomic variations in cervical disease
Integrated human papillomavirus type 16 (HPV-16) viral loads are currently estimated by quantification with real-time PCR of HPV-16 E6 (RT-E6 and HPV-16 PG) 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 (median=3.23, range=1.92-3.49) or Asian-American (median=3.78, range=1.47-37) isolates were greater than those obtained with European isolates (median=1.02, range=0.64-1.80; P<0.02 for each comparison). The distribution of HPV-16 E2 copies measured in 139 samples with RT-E2-2 (median=6150) and RT-E2-1 (median=8960) 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)=2.17, 95 % confidence interval (CI)=1.11-4.23], episomal (OR=2.14, 95 % CI=1.09-4.19), but not for HPV-16 integrated viral load (OR=1.71, 95 % CI=0.90-3.26), 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, P=0.24) or CIN-1 (5 of 14, P=0.50). HPV-16 E2 polymorphism was a significant factor that influenced measures of HPV-16 integrated viral load.
Analysis of self-collected swab samples from the genital tract could improve accrual and retention of women in studies of human papillomavirus (HPV) infection and precancerous cervical lesions. Self-collected vaginal swab specimens and physician-collected cervical swab specimens were compared for detection and typing of HPV DNA in 158 HIV-seropositive women.Paired samples were collected for 157 participants. Beta-globin was not detected in 6 (3.3%) physician-collected specimens and 8 (4.3%) self-obtained specimens collected from 11 women, leaving 146 paired samples suitable for PCR analysis. HPV DNA was amplified with the HPV primers PGMY09 and PGMY11 and typed using the line blot assay.HPV DNA was detected more frequently in self-collected samples (95 [65.1%] of 146), compared with physician-collected samples (78 [53.4%] of 146) (P = .04). Self-collected samples contained a greater number of types (mean +/- SD, 1.60 +/- 1.80 types; 95% confidence interval [CI], 1.31-1.90), compared with physician-collected samples (mean +/- SD, 1.25 +/- 1.66 types; 95% CI, 0.98-1.52) (P = .04). A good agreement between sampling methods was achieved for detection of any HPV DNA (kappa = 0.73; 95% CI, 0.58-0.89), high-risk types (kappa = 0.84; 95% CI, 0.68-0.99), and low-risk types (kappa = 0.71; 95% CI, 0.67-0.75). Agreement between sampling methods for detection of HPV DNA was found for 24 (88.8%) of 27 follow-up samples collected from a total of 20 women. A comparison of samples collected at consecutive visits revealed agreements for detection of any HPV DNA, detection of high-risk HPV, and HPV typing results between visits of 88.9% (24 of 27 samples), 81.5% (22 of 27), and 55.5% (15 of 27), respectively, for physician-collected samples, and 96.3% (26 of 27 samples), 92.6% (25 of 27), and 55.5% (15 of 27), respectively, for self-collected samples.Analysis of self-collected vaginal swab samples improved the detection rate of HPV, suggesting that such samples might be of greater value than physician-obtained samples in studies of HPV transmission.
HIV-seropositive women are at increased risk for human papillomavirus (HPV) infection, which causes high-grade squamous intraepithelial lesions (HSILs). HPV-52 is a frequent HPV type in Canadian HIV-seropositive women. Because variations of the capsid gene, designated the L1 gene, could elicit immune responses that result in different efficiencies in eliminating HPV, we described HPV-52 polymorphism and assessed whether it was associated with HPV-52 persistence in 114 women at risk or infected by HIV. Nonsynonymous variations were more frequent in the 5 putative hypervariable regions (exposed loops of L1 protein) (10 [3.2%] variations over 311 nucleotides) than in nonvariable regions (4 [0.3%] variations over 1278 nucleotides; P < 0.0001). Synonymous variations were distributed evenly between hypervariable regions (10 [3.2%] variations over 311 nucleotides) and nonvariable regions (46 [3.6%] variations over 1278 nucleotides; P = 0.88). Nonprototype (nonreference) L1 variants were detected more frequently in women of African descent (24 [60.0%] of 40 women) than in white women (23 [37.1%] of 62 women, odds ratio = 2.54, 95% confidence interval: 1.11 to 5.81; P = 0.03). In contrast to previous findings that polymorphism in the long control region (LCR) was associated with HPV-52 persistence, L1 capsid variations were not associated with persistence (P = 0.45). L1 variations are unlikely to predispose to HPV-52 persistence and thus do not help to identify women at greater risk for HSILs.
