Malignant transformation of canine oral papillomavirus (CPV1)-associated papillomas in dogs: An emerging concern?

Canine oral papillomavirus (CPV1, also known as COPV), the most common cause of non-neoplastic papillomas, has not been shown to cause squamous cell carcinomas (SCC). Furthermore, malignant transformation of benign papillomas to SCC has only been reported in a single group of dogs with severe combined immunodeficiency infected with CPV2. Here, we report a series of 7 dogs with benign CPV1-associated papillomas with histologic evidence of CPV1 causing malignant transformation to carcinoma in situ and ultimately SCC. Expression of p53 and p16 proteins in CPV1-infected cells within the benign papillomas and lesions that progressed into SCC also supported an association between papillomavirus and malignant transformation. Moreover, our retrospective analysis indicated that while there have been increased numbers of viral papillomas with malignant transformation, the number of annually diagnosed canine viral papillomas has remained constant over the past decade in our laboratory. We speculate that either an altered host immunity from increased usage of immunosuppressive drugs or changing environmental factors, e.g. increase exposure to UV radiation, may cause an increased oncogenic potential of this "low-risk" virus. This study aims to raise awareness of the malignant potential of CPV1 and to encourage further investigations into the cause of this suspected change in its oncogenic potential.

Genomic characterisation of canine papillomavirus type 17, a possible rare cause of canine oral squamous cell carcinoma.

Squamous cell carcinomas (SCCs) are the second most common cancer of the canine oral cavity resulting in significant morbidity and mortality. Recently a dog with multiple oral SCCs that contained a novel papillomavirus (PV) was reported. The aim of the present study was to determine the genome of this novel PV. To do this a short section of PV DNA was amplified from an oral SCC and 'back-to-back' primers were designed. Due to the circular nature of PV DNA, these primers were then used to amplify the remainder of the genome by inverse PCR. The PCR product was sequenced using next generation sequencing and the full genome of the PV, consisting of 8007 bp, was assembled and analysed. As this is the seventeenth PV identified from the domestic dog, the novel PV was designated Canis familiaris papillomavirus (CPV) type 17. Similar to other CPV types, the putative coding regions of CPV-17 were predicted to produce 5 early and 2 late proteins. Phylogenetic analysis of ORF L1 revealed greater than 70% similarity to CPV-2 and CPV-7 and we propose that CPV-17 also be classified as a Taupapillomavirus 1. While it appears CPV-17 is only rarely present in canine oral SCCs, evidence suggests that this PV could influence the development of oral SCCs in this species.

Molecular and immunohistochemical studies do not support a role for papillomaviruses in canine oral squamous cell carcinoma development.

Oral squamous cell carcinomas (OSCCs) are common neoplasms of dogs and are of unknown cause. Whereas papillomaviruses (PVs) are an established cause of human OSCCs, few studies have investigated canine OSCCs for a PV aetiology. In humans, a PV aetiology can be determined by detecting PV DNA and PV-induced increased p16(CDKN2A) protein (p16) within the OSCC. In this study, PCR, using four different primer sets and p16 immunohistochemistry, was used to evaluate 28 canine OSCCs for a possible PV aetiology. None of the primers amplified PV DNA from any of the OSCCs although four neoplasms contained intense p16 immunostaining. Intense p16 immunostaining would indicate a PV aetiology in a human OSCC but the absence of PV DNA suggests that the increase in p16 was not due to PV infection. Overall the results indicated that PVs are not a significant cause of canine OSCCs.

Canine papillomatosis: a retrospective study of 24 cases (2001-2011) and immunohistochemical characterization / Papilomatose em cães: estudo retrospectivo de 24 casos (2001-2011) e caracterização imuno-histoquímica

A retrospective study of 24 cases of papillomas in dogs was performed from January 2001 to March 2011. Additionally, immunohistochemistry (IHC) was used to characterize and evaluate the samples. We found that disease was observed more in mixed breed dogs, ages ranging from 6 months to 10 years (mean 3.1 years), and there was no gender predilection. The main lesion sites were the skin (75%), lips (16.7%), and eyelids (8.3%). Upon histological evaluation, we observed papillary exophytic proliferation of squamous epithelium and papillary endophytic proliferation (inverted) in 87.5% and 12.5% of cases, respectively. The tumors were characterized by spinous layer hyperplasia (87.5%) with koilocytes (70.8%) and intranuclear pale basophilic inclusions bodies (8.3%), prominent granular layer with large amounts of keratohyalin granules (95.8%), and hyperkeratosis in the stratum corneum (100%). Positive immunostaining for Papillomavirus was found in 83.3% of cases, which were distributed between the granular layer and the stratum corneum. These findings indicate the following: that papillomas in dogs are caused by Papillomavirus, the viral cytopathic effect induces epithelial lesions, viral particles are found inside the cell nuclei, and inclusions bodies are rare.

Foi realizado um estudo retrospectivo de 24 casos de papilomas em cães diagnosticados no período de janeiro 2001 a março de 2011, bem como a sua caracterização imuno-histoquímica (IHQ). Cães sem raça definida foram os mais afetados, a idade média foi de 3,1 anos, com variação de 6 meses a 10 anos e não houve predileção sexual. Quanto à localização das lesões, 75,0% estavam na pele, 16,7% no lábio e 8,3% em pálpebra. Na avaliação histológica havia proliferação papilar exofítica do epitélio escamoso em 87,5% e papilar endofítica (invertido) em 12,5%. O tumor era caracterizado por hiperplasia do estrato espinhoso (87,5%) com coilócitos (70,8%) e inclusões intranucleares basofílicas pálidas (8,3%); o estrato granular estava proeminente com grande quantidade de grânulos de querato-hialina (95,8%); e havia hiperqueratose do estrato córneo (100%). Na avaliação IHQ para Papillomavirus houve marcação nos estratos granuloso e córneo em 83,3%. Estes achados indicam que os papilomas em cães são causados por Papillomavirus, as lesões epiteliais são decorrentes do efeito citopático viral, as partículas virais estão no núcleo das células e corpúsculos de inclusão são raros.

Characterization of canine oral papillomavirus by histopathological and genetic analysis in Korea.

In August 2008, forty dogs out of 400 developed oral warts in a breeding farm in Korea. Canine oral papilloma infection is a common disease in dogs. However, there has been no report of an outbreak of canine oral papillomavirus (COPV) in a group of dogs or in dog breeding farms in Korea, and the genetic analysis of COPV in Korea has yet to be performed. This study diagnosed canine oral papilloma from the oral samples of these dogs based on histopathological examination and immunohistochemistry. Polymerase chain reaction was applied to amplify the corresponding products using preexisting primer sets for COPV and a universal human papillomavirus targeting L1 gene. Further genetic analysis of the major viral capsid gene L1 confirms the sequences of Korean COPV, which shows a close relationship to previously reported COPV. This study describes the histopathological and immunohistochemical characteristics of canine oral papilloma in a group of breeding dogs in Korea and discloses the complete L1 gene sequences of Korean COPV.

Ancient papillomavirus-host co-speciation in Felidae.

BACKGROUND: Estimating evolutionary rates for slowly evolving viruses such as papillomaviruses (PVs) is not possible using fossil calibrations directly or sequences sampled over a time-scale of decades. An ability to correlate their divergence with a host species, however, can provide a means to estimate evolutionary rates for these viruses accurately. To determine whether such an approach is feasible, we sequenced complete feline PV genomes, previously available only for the domestic cat (Felis domesticus, FdPV1), from four additional, globally distributed feline species: Lynx rufus PV type 1, Puma concolor PV type 1, Panthera leo persica PV type 1, and Uncia uncia PV type 1. RESULTS: The feline PVs all belong to the Lambdapapillomavirus genus, and contain an unusual second noncoding region between the early and late protein region, which is only present in members of this genus. Our maximum likelihood and Bayesian phylogenetic analyses demonstrate that the evolutionary relationships between feline PVs perfectly mirror those of their feline hosts, despite a complex and dynamic phylogeographic history. By applying host species divergence times, we provide the first precise estimates for the rate of evolution for each PV gene, with an overall evolutionary rate of 1.95 x 10(-8) (95% confidence interval 1.32 x 10(-8) to 2.47 x 10(-8)) nucleotide substitutions per site per year for the viral coding genome. CONCLUSION: Our work provides evidence for long-term virus-host co-speciation of feline PVs, indicating that viral diversity in slowly evolving viruses can be used to investigate host species evolution. These findings, however, should not be extrapolated to other viral lineages without prior confirmation of virus-host co-divergence.