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.
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.
Azizi
Naoufel
N
Laboratoire de Virologie Moléculaire, Centre de Recherche du Centre Hospitalier de l'Université de Montréal, Montréal, Québec, Canada.
Brazete
Jessica
J
Laboratoire de Virologie Moléculaire, Centre de Recherche du Centre Hospitalier de l'Université de Montréal, Montréal, Québec, Canada.
Hankins
Catherine
C
Department of Epidemiology, Biostatistics and Occupational Health Medicine, McGill University, Montreal, Québec, Canada.
Money
Deborah
D
Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, BC, Canada.
Fontaine
Julie
J
Laboratoire de Virologie Moléculaire, Centre de Recherche du Centre Hospitalier de l'Université de Montréal, Montréal, Québec, Canada.
Koushik
Anita
A
Department of Epidemiology, Biostatistics and Occupational Health Medicine, McGill University, Montreal, Québec, Canada.
Rachlis
Anita
A
Department of Medicine, Sunny Brook Health Science Centre, University of Toronto, Toronto, Ontario, Canada.
Pourreaux
Karina
K
Department of Epidemiology, Biostatistics and Occupational Health Medicine, McGill University, Montreal, Québec, Canada.
Ferenczy
Alex
A
Department of Pathology and Obstetrics & Gynecology, The Sir Mortimer B. Davis-Jewish General Hospital and McGill University, Québec, Canada.
Franco
Eduardo
E
Departments of Oncology, Division of Epidemiology, McGill University, Montreal, Québec, Canada.
François Coutlée For The Canadian Women's Hiv Study Group
Département de Microbiologie et Immunologie, Université de Montréal, Montréal, Québec, Canada.|Departments of Oncology, Division of Epidemiology, McGill University, Montreal, Québec, Canada.|Laboratoire de Virologie Moléculaire, Centre de Recherche du Centre Hospitalier de l'Université de Montréal, Montréal, Québec, Canada.
D004279:DNA, Viral / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D005260:Female; D005843:Geography; D015658:HIV Infections / Q000150:complications; D052162:Human papillomavirus 16 / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D010957:Plasmids; D011110:Polymorphism, Genetic; D011533:Proviruses / Q000235:genetics; D044364:Vaginal Douching; D016662:Virus Integration; D002578:Uterine Cervical Dysplasia / Q000821:virology*
D004279:DNA, Viral; D004268:DNA-Binding Proteins; C054855:E2 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral