MULTIFOCAL VULVAR INTRAEPITHELIAL NEOPLASIA GRADE-III AND MULTICENTRIC LOWER GENITAL-TRACT NEOPLASIA IS ASSOCIATED WITH TRANSCRIPTIONALLY ACTIVE HUMAN PAPILLOMAVIRUS

Background. The incidence of vulvar intraepithelial neoplasia Grade II I (VIN III) is increasing and is diagnosed at a younger age than previ ously. VIN III is often multifocal and frequently coexists with multic entric dysplastic lesions in the cervix and vagina. Warty-type VIN III more often has been found to contain human papillomavirus (HPV) DNA t han basaloid-type VIN III. The authors performed HPV DNA polymerase ch ain reaction (PCR) analysis in 48 VIN III biopsies and reverse transcr iptase (RT)-PCR in 8 HPV-16 DNA-positive multifocal VIN III biopsies t o detect E6/E7 transcripts. Methods. Human papillomavirus DNA detectio n and histologic analysis were performed on alternating slides of para ffin embedded biopsies. Polymerase chain reaction was performed with c onsensus primers, and HPV typing was performed by direct sequencing. T otal RNA was isolated from frozen biopsies by centrifuging a guanidini um thiocyanate (GTC) lysate through a cesium chloride (CsCl) cushion. The RT reaction was performed using a 3' primer, located just downstre am of the E7 stop codon, and the PCR reaction was performed using the same 3' primer and a 5' primer located just downstream of the E6 start codon. Results. The mean age of the 48 patients was 37.7 years. Eight y-one percent had multifocal VIN III. Sixty-six percent had multicentr ic neoplasia. Forty-six percent of the biopsies were warty-type, 17% b asaloid-type, 35% mixed-type and 2% differentiated-type. Ninety-two pe rcent were HPV-positive and 83% contained HPV-16 DNA. Human papillomav irus DNA was more often present in multifocal VIN III lesions than in unifocal VIN III lesions and also more often in VIN III lesions coexis ting with other dysplastic multicentric lesions than in unicentric VIN III lesions. Warty-type VIN III more often contained koilocytes than basaloid-type VIN III. A correlation between different morphologic for ms of VIN III and the presence of HPV DNA was not found. Both types of VIN III often coexist in one lesion. In all the RT-PCRs, a 593-base-p air fragment was detected, corresponding to the expected length of the major E6-E7 mRNA. Conclusions. The observed high prevalence of trans criptionally active HPV DNA associated with multifocal and multicentri c dysplasia suggests a role of HPV in the pathogenesis of these lesion s. A positive correlation between different morphologic forms of VIN I II and the presence of HPV DNA was not found.