Short Communication Increased Risk for Cervical Disease Progression of French Women Infected with the Human Papillomavirus Type 16 E6-350G Variant Martha Grodzki, 1 Guillaume Besson, 2 Christine Clavel, 2 Annie Arslan, 3 Silvia Franceschi, 3 Philippe Birembaut, 2 Massimo Tommasino, 3 and Ingeborg Zehbe 4 1 Johannes Gutenberg University, Medical Microbiology, Mainz, Germany; 2 Laboratoire Pol Bouin, Hopital Maison Blanche, CHU, Reims, France; 3 IARC, Lyon, France; and 4 Thunder Bay Regional Health Sciences Centre, Regional Cancer Care, Thunder Bay, Ontario, Canada Abstract To test the significance of human papillomavirus (HPV) type 16 and HPV16 E6 variants as risk factors for viral persistence and progression to high-grade lesion, we did a nested case-control study within a cohort study of >15,000 Caucasian French women. Three groups infected with high- risk HPV were compared: (a ) women with cleared infection (controls, n = 201), (b ) women with persistent infection (cases, n = 87), and (c ) women who progressed into high- grade lesion (cases, n = 58). Women with persistent HPV infection and those that progressed into high-grade lesions were likelier to harbor HPV16 than other high-risk HPV types [odds ratio (OR), 2.4; 95% confidence interval (95% CI), 1.3-4.3 and OR, 4.2; 95% CI, 2.2-8.1, respectively]. Notably, especially elevated ORs of persistence (3.0; 95% CI, 1.4-6.7) and progression (6.2; 95% CI, 2.7-14.3) were found among women who harbored the HPV16 350G variant. Thus, HPV type and HPV16 variant seem to be risk factors for viral persistence and progression of infections into high-grade cervical lesions. (Cancer Epidemiol Biomarkers Prev 2006; 15(4):820–2) A subgroup of human papillomaviruses (HPV), referred to as high-risk types, are the etiologic agents of cervical cancer (1). Among the high-risk types HPV16 is the most prevalent type in premalignant and malignant cervical lesions worldwide (2, 3). Persistence of viral infection has a key role in cervical cancer development (4, 5). HPV16 is likelier to persist (6) and to cause progression into cervical intraepithelial neoplasia (CIN) than other high-risk HPV types (7, 8). Nucleic acid sequencing of HPV16 genomes has revealed the existence of numerous natural variants that differ from the original European prototype sequence (9) up to 2% in the coding region and/or up to 5% in the noncoding region (10). HPV16 variants have been classified as five major phylogenetic clusters (or lineages): European, Asian, Asian American, African, and North American. For each lineage, several subclasses have been identified (summarized in ref. 11). Interestingly, three codon sites in the HPV16 E6 open reading frame coding for amino acids 10, 14, and 83 were shown to be under selective pressure (12). Variants at these residues lead to changes from arginine to glycine or isoleucine (R10G or R10I) at codon 10, from glutamine to histidine or aspartic acid (Q14H or Q14D) at codon 14, and from leucine to valine (L83V) at codon 83. Recent studies suggest that HPV16 E6 variants are involved in determining persistence of the viral infection and the development of cervical lesions (13-20). Some of these have shown that non-European HPV16 variants are more associated with disease progression than the European variants (21, 22). By using cross-sectional analyses of several European populations, we have previously shown that within the European lineage, the L83V E6 variant harboring a nucleotide substitution at position 350 (HPV16 350G) alone or in combination with other polymorphisms is more prevalent in cervical cancer than in CIN (23-25). This phenomenon may be explained by the fact that natural variants alter the immunogenic and/or carcinogenic properties of the virus. In this study, we have used a large French cohort to elucidate further whether HPV16 and HPV16-specific variants are predictors for persistence of infection and progression into high-grade lesion. We did a nested case-control study within a large cohort study of >15,000 Caucasian French women ages between 18 and 76 years (median = 34 years). They had been recruited between 1997 and 2003 among women who underwent their biennial or triennial routine screening in the Department of Obstetrics and Gynaecology at the CHU in Reims, France (26). The follow-up ranged between 12 to 72 months (median = 34 months; ref. 26). All women were informed of the aim of the program and gave their written consent. This study was approved by the local ethical committee. All cohort women were tested for the presence of the high-risk HPV types by the Hybrid Capture II assay (Digene, Gaithersburg, MD), and only women who were positive for high-risk HPV types and did not have high-grade lesions at baseline were eligible for the present study. Women were excluded if they had a cytologic abnormality and/or an untreated cervical lesion in the past 2 years, were pregnant, or had an HIV/AIDS diagnosis. To identify the high-risk type involved, HPV16 was tested by PCR using E6 specific primers, whereas other high-risk HPV types were tested by the Linear Blot Array 820 Cancer Epidemiol Biomarkers Prev 2006;15(4). April 2006 Received 11/8/05; revised 1/6/06; accepted 2/2/06. Grant support: Deutsche Krebshilfe, Bonn, Germany grant 70-3050-Ze (I. Zehbe); La Ligue Contre le Cancer (M. Tommasino); and Comite ´ du Rho ˆne et Comite ´ de la Dro ˆme, France (M. Tommasino). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Note: This work has been done in fulfillment of the requirements for the M.D. thesis of M. Grodzki. M. Grodzki and G. Birembaut contributed equally to this work. Requests for reprints: Ingeborg Zehbe, Thunder Bay Regional Health Sciences Centre, Regional Cancer Care Programme, 980 Oliver Road, Thunder Bay, Ontario P7B 6V4, Canada. Phone: 807-684-7256; Fax: 807-684-5806. E-mail: zehbei@tbh.net Copyright D 2006 American Association for Cancer Research. doi:10.1158/1055-9965.EPI-05-0864 Research. on January 31, 2016. © 2006 American Association for Cancer cebp.aacrjournals.org Downloaded from Research. on January 31, 2016. © 2006 American Association for Cancer cebp.aacrjournals.org Downloaded from Research. on January 31, 2016. © 2006 American Association for Cancer cebp.aacrjournals.org Downloaded from