Microarray analysis of canine papillomavirus lesions

Papillomaviren verursachen eine breite Palette von Krankheitsbildern bei Mensch und Tier, welche von asymptomatischen Infektionen, gutartigen Warzen, pigmentierten Plaques, in situ Karzinomen bis zu invasiven Plattenepithelkarzinomen reichen. Bei Hunden sind die gesamten genomischen Sequenzen von dreizehn verschiedenen Papillomaviren beschrieben, die mit unterschiedlichen klinischen Erscheinungsbildern verbunden sind. Allerdings können weder die phylogenetische Klassifizierung eines Hundepapillomavirus (CPV) noch das Vorhandensein von unterschiedlichen Genregionen genau den Verlauf einer Infektion auf individueller Ebene vorhersagen. In der vorliegenden Studie führten wir eine Microarray-Analyse von CPV assoziierten Läsionen (CPV1, CPV3, CPV5) durch. Die Expression von sechsundneunzig Genen war signifikant mindestens 4-fach rauf oder runter reguliert. Weiter haben wir die RNA Levels von KLK8, MCM5, ORC1 und PAX6, die als Biomarker in der Humanmedizin diskutiert werden in Papillomavirus Läsionen untersucht. Wir konnten durch qRT-PCR die erhöhte Expression dieser Gene bestätigen. Papillomaviruses induce a wide range of clinical conditions in humans and animals, ranging from asymptomatic infection, benign warts, pigmented plaques to in situ carcinoma and invasive squamous cell carcinomas. In dogs, entire genomic sequences of thirteen different papillomaviruses are described so far and that appear to be associated with different clinical presentations. However, neither the phylogenetic classification of a canine papillomavirus (CPV) nor the presence of distinct motifs on the genome can unanimously predict the fate of an infection on the individual level. In the present study we performed a microarray analysis of CPV associated lesions (CPV1, CPV3, CPV5). Ninety-six genes were at least 4 fold up or down-regulated on a significant level. We further investigated in CPV lesions, the RNA levels of KLK8, MCM5, ORC1, and PAX6, which are discussed as biomarkers in the human medicine. We could confirm the up regulation of these genes by qRT PCR

Novel snake papillomavirus does not cluster with other non-mammalian papillomaviruses

Papillomaviruses (PVs) are associated with the development of neoplasias and have been found in several different species, most of them in humans and other mammals. We identified, cloned and sequenced PV DNA from pigmented papilloma-like lesions of a diamond python (Morelia spilota spilota). This represents the first complete PV genome discovered in a Squamata host (MsPV1). It consists of 7048 nt and contains the characteristic open reading (ORF) frames E6, E7, E1, E2, L1 and L2. The L1 ORF sequence showed the highest percentage of sequence identities to human PV5 (57.9%) and Caribbean manatee (Trichechus manatus) PV1 (55.4%), thus, establishing a new clade. According to phylogenetic analysis, the MsPV1 genome clusters with PVs of mammalian rather than sauropsid hosts

Complete canine papillomavirus life cycle in pigmented lesions

Canine papillomaviruses (CPVs) have been identified in various benign and malignant neoplastic skin disorders. The most frequent manifestations of CPV infections are classical warts and pigmented plaques. Although the etiology of canine oral papillomatosis is well established, knowledge about CPVs role in the development of pigmented plaques remains vague. Indeed, as CPV DNA may frequently be found on clinically healthy canine skin, its mere detection in lesions cannot be regarded as a sufficient indicator of causality. Whether CPVs are actually active in pigmented plaques, a requirement for any conceivable involvement, is consequently an open question. To enquire such viral activity, two distinct clinical cases of canine pigmented lesions were evaluated in greater detail. The histological findings in the two cases supported the clinical diagnosis of pigmented viral plaques. Sequencing of amplified DNA from these lesions revealed the genomes of two novel CPV types, i.e. CPV9 and CPV14, both putatively belonging to the genus Chi. Furthermore, transcription and splicing of corresponding CPV mRNA could be shown by RT-PCR in the respective lesions. Finally, viral particles were detected by electron microscopy in homogenates as well as in nuclei of keratinocytes in pigmented lesions. In conclusion, the results link clinical signs of pigmented plaques to histological changes, the presence of CPV specific DNA, viral gene transcription, and the presence of viral particles in and from the lesions. Thus, the findings outline the entire replicative cycle of CPVs in pigmented plaques, which might help understanding the relationship between these viruses and the associated disorders