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.
Laboratoire de Virologie Moléculaire, Centre de Recherche du Centre Hospitalier de l'Université de Montréal, Montréal, Québec, Canada.
Naoufel
N
Azizi
Laboratoire de Virologie Moléculaire, Centre de Recherche du Centre Hospitalier de l'Université de Montréal, Montréal, Québec, Canada.
Jessica
J
Brazete
Department of Epidemiology, Biostatistics and Occupational Health Medicine, McGill University, Montreal, Québec, Canada.
Catherine
C
Hankins
Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, BC, Canada.
Deborah
D
Money
Laboratoire de Virologie Moléculaire, Centre de Recherche du Centre Hospitalier de l'Université de Montréal, Montréal, Québec, Canada.
Julie
J
Fontaine
Department of Epidemiology, Biostatistics and Occupational Health Medicine, McGill University, Montreal, Québec, Canada.
Anita
A
Koushik
Department of Medicine, Sunny Brook Health Science Centre, University of Toronto, Toronto, Ontario, Canada.
Anita
A
Rachlis
Department of Epidemiology, Biostatistics and Occupational Health Medicine, McGill University, Montreal, Québec, Canada.
Karina
K
Pourreaux
Department of Pathology and Obstetrics & Gynecology, The Sir Mortimer B. Davis-Jewish General Hospital and McGill University, Québec, Canada.
Alex
A
Ferenczy
Departments of Oncology, Division of Epidemiology, McGill University, Montreal, Québec, Canada.
Eduardo
E
Franco
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.
François Coutlée For The Canadian Women's Hiv Study Group
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