Molecular epidemiology of HPV in cutaneous and mucosal epithelia from individuals with different immunological status

ELISA M. BOLATTI; DIEGO CHOUHY; PABLO E. CASAL; EMMA J. STELLA; ADRIANA A. GIRI

Rosario

Workshop; First ICGEB Workshop Human Papillomavirus; 2014

Human Papillomaviruses (HPVs) are DNA tumour viruses infecting mucosal and cutaneous epithelia. More than 190 HPV types have been described and divided into five genera (alpha, beta, gamma, nu, and mu) according to their phylogenetic relationships. Mucosal HPV infection is a well-established cause of cervical cancer and is a relevant factor in other anogenital and head and neck cancers. On the other hand, the clinical implications of the cutaneous HPV types are still unknown and it has been proposed that immunosuppression (UV + immunosuppressive treatment) may contribute to HPV infection. In order to explore the diversity of mucosal and cutaneous HPVs from individuals with different immunological status, we designed a transversal descriptive study. Cervical cells samples were obtained from 117 women with cervical pathological diagnosis and 11 immusuppressed women (HIV positive). All samples were analyzed with the L1HPVPCR assay using the MY consensus primers and by hanging droplet PCR with the CUT primer system. HPV DNA was present in 101/128 samples (79%), 75 (74%) of them contained at least one carcinogenic HPV type, being HPV-16 (alpha-9) the most prevalent type. L1HPVPCR assay and CUT primers system had a general concordance of 58 % (74/128). HPV types were determined by hybridization to type-specific probes (L1HPVPCR) or by direct sequencing (CUT system). The same HPV type was found in 39 women using both test, leading to a type-specific concordance of 38% (39/101). Skin samples were collected from the foreheads of 100 immunocompetent (IC) and 64 immunosuppressed (IS) individuals and analyzed by hanging droplet PCR with the FAP primer system. HPV DNA was more frequently found in the IC group (65% IC vs. 56% IS). In both populations, most of HPV types and putative types identified belonged to the genera beta- and gamma-PV, being HPV-20 (beta-1) the most prevalent type. Time of sun exposure and age were significantly higher in IC HPV-infected subjects, compared to non HPV-infected subjects. Using hanging droplet long PCR assay, the full-length genomes of two gamma-PV putative types were amplified. Novel putative types EP1 and FA91 showed 85% and 73% of sequence identity in L1 ORF to the HPV-173 (gamma-1) and HPV-148 (gamma-12), respectively. These findings expand our knowledge about diversity, evolution and medical implications of the family Papillomaviridae.