One virus, one lesion—individual components of CIN lesions contain a specific HPV type

Abstract

In 20-40% of cervical intra-epithelial neoplasia (CIN) and in 4-8% of cervical carcinoma tissue specimens, multiple HPV genotypes have been detected. Whole tissue section (WTS) PCR does not determine how the individual types relate causally to complex and multiple CIN. Our objective was to determine whether laser capture micro-dissection (LCM) with HPV PCR genotyping (LCM-PCR) could accurately recover type-specific HPV DNA from epithelial cells in individual areas of CIN and normal epithelium, and whether one or more viruses are present in one lesion. For that, histologically selected samples of CIN and normal epithelium were isolated by LCM and analysed by the SPF(10) PCR/LiPA(25) (version 1) HPV genotyping system for 25 HPV genotypes. HPV genotypes detected in 756 areas of CIN (grade 1, 2 or 3) by LCM-PCR were compared with results obtained by WTS-PCR in 60 cases (74 biopsies). We showed that when a single HPV type is detected by WTS-PCR, that type was almost always (94%; 29/31) recovered by LCM-PCR from CIN. When multiple HPV types were present by WTS-PCR, their distribution within histological sections could be mapped by LCM-PCR. Association of a single HPV type with a discrete area of CIN was found for 93% (372/399) of LCM fragments analysed by PCR. We found colliding CIN lesions associated with separate HPV types and only 62% (61/99) of HPV types detected by WTS-PCR were found in CIN by LCM-PCR. Therefore, the LCM-PCR technique was found very accurate for high-resolution HPV genotyping and for assigning an individual HPV type to an area of CIN. At LCM level, in cervical biopsy sections with multiple HPV infections, the relation between HPV types and CIN lesions is often complex. Almost every HPV type found in CIN by LCM-PCR is associated with a biological separate independent CIN lesion-one virus, one lesion.

Biological activity of probable/possible high‐risk human papillomavirus types in cervical cancer

“…This question may be further clarified by separate analysis of different lesion regions isolated by laser microdissection as recently described for CIN lesions. 40 At present, the current data show that absence of transcription for majority of the types found as DNA co-infection (74%) helps to eliminate their causal role in tumor development in the face of multiple infections. These types may represent early, regressing or latent infections residing in adjacent normal mucosa, stroma or precancer lesions.…”

Frequency of twelve carcinogenic human papilloma virus types among women from the South Backa region, Vojvodina, Serbia

“…Some more recent research investigated the importance of multiple infections for carcinogenesis. Quint et al examined the distribution of multiple HPV infections in CIN lesions by the laser capture microdissection PCR method and concluded that every HPV type found in CIN is associated with a biologically separate and independent CIN lesion (38).…”

Multiple Human Papillomavirus Infections with High Viral Loads Are Associated with Cervical Lesions but Do Not Differentiate Grades of Cervical Abnormalities

“…In the present study, this is supported by the high variety of different HPV genotypes found with high viral loads in a single sample. We assume that these infections are biologically independent (34) and that multiple infections might be the result of an impaired immune system and/or frequent changes in sexual partners. While multiple high viral loads are significantly associated with cervical cancer precursors, it is very unlikely that multiple HPV types are contributing to the same lesion and should thus be considered true risk factors.…”