title: Linear viral load increase of a single HPV-type in women with multiple HPV infections predicts progression to cervical cancer.
abstract: Persistent high-risk human papillomavirus (HPV) infection is strongly associated with development of high-grade cervical intraepithelial neoplasia or cancer (CIN3+). In single type infections, serial type-specific viral-load measurements predict the natural history of the infection. In infections with multiple HPV-types, the individual type-specific viral-load profile could distinguish progressing HPV-infections from regressing infections. A case-cohort natural history study was established using samples from untreated women with multiple HPV-infections who developed CIN3+ (n = 57) or cleared infections (n = 88). Enriched cell pellet from liquid based cytology samples were subjected to a clinically validated real-time qPCR-assay (18 HPV-types). Using serial type-specific viral-load measurements (≥3) we calculated HPV-specific slopes and coefficient of determination (R(2) ) by linear regression. For each woman slopes and R(2) were used to calculate which HPV-induced processes were ongoing (progression, regression, serial transient, transient). In transient infections with multiple HPV-types, each single HPV-type generated similar increasing (0.27copies/cell/day) and decreasing (-0.27copies/cell/day) viral-load slopes. In CIN3+, at least one of the HPV-types had a clonal progressive course (R(2)  ≥ 0.85; 0.0025copies/cell/day). In selected CIN3+ cases (n = 6), immunostaining detecting type-specific HPV 16, 31, 33, 58 and 67 RNA showed an even staining in clonal populations (CIN3+), whereas in transient virion-producing infections the RNA-staining was less in the basal layer compared to the upper layer where cells were ready to desquamate and release newly-formed virions. RNA-hybridization patterns matched the calculated ongoing processes measured by R(2) and slope in serial type-specific viral-load measurements preceding the biopsy. In women with multiple HPV-types, serial type-specific viral-load measurements predict the natural history of the different HPV-types and elucidates HPV-genotype attribution.
authors.affiliation: Department of Clinical and Molecular Pathology, AML, Sonic Healthcare, Antwerp, Belgium.
authors.forename: Christophe E
authors.identifier: 0000-0003-3462-6557
authors.initials: CE
authors.lastname: Depuydt
authors.affiliation: Laboratory for Cell Biology and Histology, University of Antwerp, Antwerp, Belgium.
authors.forename: Sofie
authors.initials: S
authors.lastname: Thys
authors.affiliation: Department of Clinical and Molecular Pathology, AML, Sonic Healthcare, Antwerp, Belgium.|Intermediate Structure for Human Body Material, AML, Sonic Healthcare, Antwerp, Belgium.
authors.forename: Johan
authors.initials: J
authors.lastname: Beert
authors.affiliation: Department of Clinical and Molecular Pathology, AML, Sonic Healthcare, Antwerp, Belgium.
authors.forename: Jef
authors.initials: J
authors.lastname: Jonckheere
authors.affiliation: Department of Clinical and Molecular Pathology, AML, Sonic Healthcare, Antwerp, Belgium.
authors.forename: Geert
authors.initials: G
authors.lastname: Salembier
authors.affiliation: Department of Clinical and Molecular Pathology, AML, Sonic Healthcare, Antwerp, Belgium.|Laboratory for Cell Biology and Histology, University of Antwerp, Antwerp, Belgium.
authors.forename: Johannes J
authors.initials: JJ
authors.lastname: Bogers
mesh_terms: D000328:Adult; D060085:Coinfection; D018450:Disease Progression; D005260:Female; D006801:Humans; D027383:Papillomaviridae / Q000145:classification* / Q000502:physiology*; D030361:Papillomavirus Infections / Q000821:virology*; D012367:RNA, Viral / Q000235:genetics; D002583:Uterine Cervical Neoplasms / Q000821:virology*; D019562:Viral Load; D002578:Uterine Cervical Dysplasia / Q000821:virology*
chemical_list: D012367:RNA, Viral
keywords: cervical cancer screening; cervical intraepithelial neoplasia; clonal; liquid-based cytology leftover; real-time quantitative PCR; transient virion producing; viral doubling time