Feline Papillomavirus Research Articles

Classification and clinical significance of Papillomavirus infection in domestic cats

During the last decades, the infection with papillomavirus in domestic cats gained interest from the veterinary community due to its significant impact on the companion's animal's health. Therefore, in this review, we aim to present a concise classification of feline papillomaviruses and their clinical relevance in domestic felines. Initially, the different types of papillomaviruses affecting domestic cats are described. Here, we emphasize the molecular diversity and transmission ways to better understand each virus type and its clinical implications. Furthermore, we explore the clinical importance of papillomavirus infections, analyzing their various manifestations such as skin or oral lesions. We outline the signs and symptoms of these infections, shedding light on the oncogenic mechanisms used by the virus. The knowledge gained from this analysis holds the potential to refine veterinary medical practices, enabling the effective management of this condition and ultimately enhancing the overall quality of life for our feline companions.

Feline SCCs of the Head and Neck Display Partial Epithelial-Mesenchymal Transition and Harbor Stem Cell-like Cancer Cells.

Squamous cell carcinoma of the head and neck (HNSCC) is a malignant cancer disease in humans and animals. There is ample evidence that the high plasticity of cancer cells, i.e., their ability to switch from an epithelial to a mesenchymal, endothelial, and stem cell-like phenotype, chiefly contributes to progression, metastasis, and multidrug resistance of human HNSCCs. In feline HNSCC, the field of cancer cell plasticity is still unexplored. In this study, fourteen feline HNSCCs with a known feline papillomavirus (FPV) infection status were subjected to histopathological grading and subsequent screening for expression of epithelial, mesenchymal, and stem cell markers by immunohistochemistry (IHC) and immunofluorescence staining (IF). Irrespective of the FPV infection status, all tumors except one corresponded to high-grade, invasive lesions and concurrently expressed epithelial (keratins, E-cadherin, β-catenin) and mesenchymal (vimentin, N-cadherin, CD146) proteins. This finding is indicative for partial epithelial-mesenchymal transition (pEMT) events in the lesions, as similarly described for human HNSCCs. IF double staining revealed the presence of CD44/CD271 double-positive cells notably within the tumors' invasive fronts that likely correspond to cancer stem cells. Taken together, the obtained findings suggest that feline HNSCCs closely resemble their human counterparts with respect to tumor cell plasticity.

Canine and feline papillomaviruses: an update.

Papillomaviruses are small viruses able to cause disease not only in mammalians, but also in birds and reptiles. In recent years, a rising number of papillomaviruses have been identified in dogs and cats, totaling 24 canine papillomavirus (CPV) and six feline papillomavirus (FcaPV). The canine and feline papillomaviruses (CPVs and FcaPVs, respectively) are responsible for multiple lesions in these domestic species but the potential pathological relevance of some recently identified types remains to be determined. CPVs are associated with oral papillomatosis, cutaneous papillomas and viral pigmented plaques, and have been rarely associated with the development of oral and cutaneous squamous cell carcinomas in their canine hosts. FcaPVs are associated with oral papillomas, viral plaques, and Bowenoid in situ carcinomas. The present review provides readers with the more recent advances on dog and cat papillomavirus research, bringing an update on this field to both veterinary practitioners and the virology community at large.

Feline Oral Squamous Cell Carcinoma: A Critical Review of Etiologic Factors.

Simple SummaryFeline oral squamous cell carcinoma (FOSCC) is the most common oral neoplasia in cats. This malignant tumor is locally invasive, has a high mortality rate, and its etiology is not yet known. A critical review about the potential etiologic factors of FOSCC was performed, considering publications between 2000 and 2022. The initial search resulted in 553 publications, with only 26 of these being included in the review. Sixteen studies were related to viral etiology and nine related to environmental factors such as exposure to tobacco smoke, ectoparasitic products, and the presence of oral comorbidities. When evaluated, feline papillomavirus was detected in 16.2% of samples of FOSCC. In the three studies focused on exposure to tobacco smoke, 35.2% (30/85) of cats with FOSCC had a history of this exposure. Among 485 cats with FOSCC, 6.4% had dental and oral pathology. The present study demonstrates that the available evidence on the etiology of FOSCC is still limited, however, there has been increasing interest in this topic.Feline oral squamous cell carcinoma (FOSCC) is the most common oral neoplasia in cats. This malignant tumor is locally invasive, has a high mortality rate, and its etiology is not yet known. In humans, head and neck squamous cell carcinoma is associated with tobacco smoke, alcohol consumption, and human papillomavirus infection. Herein, a critical review about the potential etiologic factors of FOSCC was performed, considering publications between 2000 and 2022, aiming to synthesize all available scientific evidence regarding this issue. Recommendations of the PRISMA statement and the Cochrane Collaboration were followed and the PubMed database searched by using the MeSH terms MeSH terms “oral”, “mouth”, “lingual”, “labial”, “gingiva”, “carcinoma”, “squamous”, and “feline”. The selection process for eligible studies was based on specific inclusion and exclusion criteria and the quality of the studies assessed. The initial search resulted in 553 publications, with only 26 of these being included in the review. Sixteen studies were related to viral etiology and nine related to environmental factors such as exposure to tobacco smoke, ectoparasitic products, and the presence of oral comorbidities. When evaluated, feline papillomavirus was detected in 16.2% of samples of FOSCC. In the three studies focused on exposure to tobacco smoke, 35.2% (30/85) of cats with FOSCC had a history of this exposure. The consumption of canned food and the use of deworming collars were associated, in only one publication, with a risk of neoplasia increased by 4.7 and 5.3 times, respectively. Among 485 cats with FOSCC, 6.4% had dental and oral pathology (i.e., periodontal disease or feline chronic gingivostomatitis). The present study demonstrates that the available evidence on the etiology of FOSCC is still limited, however, there has been an increasing interest on this topic. To better understand the role of the possible etiological factors of this aggressive disease, and model for its human counterpart, large, prospective multi-institutional studies are needed.

Sublingual papillomas of cheetahs in southern Africa.

Nine distinct papillomaviruses (Lambdapapillomavirus) have been described in domestic and nondomestic cats, but not in cheetahs. These viruses have been associated with cutaneous papillomas or plaques, bowenoid in situ carcinomas, feline cutaneous squamous cell carcinomas (SCC), feline sarcoids, and oral (often sublingual) papillomas. Fourteen cheetahs from the AfriCat foundation (Namibia) and one from the Ann van Dyk Cheetah center (South Africa) presented with sublingual lesions reminiscent of sublingual papillomas. Two animals were biopsied and the histopathology revealed benign proliferative epithelial lesions with prominent thickening of the overlying squamous epithelium. Throughout the squamous epithelial layers were cells with nuclear enlargement, irregularity of the nuclear membranes and cell contours, focal hyperchromasia of the nuclei, and perinuclear halos, reminiscent of a virus-associated process as seen in papillomavirus infections. Thirteen more cheetahs were sampled and the tissue snap frozen for molecular characterization. Amplification and sequencing of the papillomavirus L1, E6, E7, and E1 gene regions was achieved with modified primers. Maximum likelihood phylogenetic analyses revealed all 15 cheetah papilloma samples were 99.99% genetically similar and closely related to, but genetically distinct from any known felinepapillomaviruses. All cheetahs were FIV and FeLV negative. The results suggest the samples identified in this study can be considered a previously undescribed or novel feline papillomavirus and the authors propose "Acinonyx jubatus papillomavirus type 1" (AjPV-1), within the Lambdapapillomavirus 1 genus (Family: Papillomaviridae).

Genome Sequence of Feline Papillomavirus Strain P20 Assembled from Metagenomic Data from the Skin of a House Cat Owner.

ABSTRACTA feline papillomavirus genome was assembled from metagenomic sequencing data collected from the skin of a house cat owner. The circular genome of strain P20 is 8,069 bp in length, has a GC content of 54.38%, and displays genome organization typical of feline papillomaviruses. The genome exhibits approximately 75% sequence similarity to other feline papillomavirus genomes.

Cutaneous papilloma associated with a novel papillomavirus sequence in a cat.

A 5-mm diameter mass developed on the nasal planum of a 4.5-y-old castrated male domestic shorthaired cat; the mass was raised ~2 mm above the surrounding skin. Histology revealed focal thickening of the epidermis with marked orthokeratosis. Many of the epidermal cells within the mass had prominent papillomavirus-induced changes. A diagnosis of a viral papilloma was made, and a DNA sequence from a novel papillomavirus type was amplified from the lesion. Although the sequence was most similar to other feline papillomavirus types, the low level of similarity was suggestive of a novel papillomavirus genus. There has been no recurrence of the mass or development of additional lesions in the 6 mo since the mass was removed. This is the third cutaneous papilloma reported in a cat; a putative feline papillomavirus type has not been identified previously within these lesions, to our knowledge. Our findings expand the range of lesions associated with papillomaviruses in cats and increase the number of papillomavirus types that infect cats.

PSIII-12 Identification and Classification of Novel Animal Associated Viruses from the Human Skin Virome of Pet Owners and Agricultural Workers

Abstract Viruses are the most abundant biological entity on earth. Despite this, viral diversity in current reference databases is poorly described. With limited number of studies investigating environmental viromes, our understanding of the virosphere composition and diversity is limited. Improvement in metagenomic sequencing methodologies has allowed for identification of novel genomic sequences, especially for new and emerging viral taxa, and improvement in existing ambiguous viral phylogenetic groupings. Here we describe a viral discovery-based study that evaluated viral metagenomes collected from the hands and scalps of 42 human subjects (n = 642) for identification of environmentally contracted DNA viruses. Due to direct interaction between humans and animals either through owning pets or through vocation, it is hypothesized that surveillance of the human skin virome will contain animal associated viruses contracted through environmental contact that can then potentially become zoonotic pathogens. To investigate this, viral genome assemblies from the metagenomes were bioinformatically predicted using model and reference-based tools. A gene and protein sequence variation and binning approach was then employed to resolve viral dark matter. Evaluation of animal associated viruses resulted in the identification of a feline papillomavirus from a subject who reportedly owned a cat. Phylogenetic analysis suggests this isolate is related to feline papillomavirus type 2 sharing approximately 75% sequence synteny thus making it a novel papillomavirus isolate. Additionally, a new grouping of porcine associated Cressdnaviricota viruses were identified through protein sequence similarity network clustering of conserved Rep genes and all other NCBI reported Rep gene sequences. This grouping contained both novel viral genomes collected from the human skin virome dataset and unclassified Cressdnaviruses all collected from porcine samples obtained from NCBI. Further phylogenetic analysis showed similarity to this novel cluster to that of porcine circovirus (PCV). This discovery and phylogenetic based investigation of novel viral genome assemblies demonstrates that animal associated viruses can be identified from the skin of humans and thus studying human viromes can provide information into the spread and emergence of novel viruses.

Feline cystadenomatosis affecting the ears and skin of 57 cats (2011-2019).

This study aimed to understand epidemiological factors associated with feline cystadenomatosis, including signalment and papillomavirus PCR status. Cystadenomatosis is an uncommon condition primarily involving the ceruminous and apocrine skin and ear glands. This was a retrospective case series. Clinical records from 2011 to 2019 from a tertiary referral hospital in Boston, MA, USA were screened for cases, and case data were re-evaluated and analyzed. The total patient pool contained 65,385 individual cats, of which 797 were referred to the dermatology service. Medical records and biopsy specimens were reviewed; the information collected included signalment, clinical signs, physical examination and diagnostic tests, comorbidities and histopathologic findings. PCR was performed on biopsy specimens to test for papillomavirus DNA. The cystadenomatosis population consisted of 57 cases (7.1% of total cases referred to the dermatology service) with 105 affected ears. Twenty-seven cases (48 ears) were confirmed via histopathology; four cats (7%) exhibited clinically cystic lesions on the periocular, periorbital and perianal regions; only one cat did not have pinnal lesions. Domestic shorthair cats were most often affected. Relative risk for cystadenomatosis was 2.24 times higher in male cats. In 48 cats (84.2%), ears were bilaterally affected. Seven cats (12.3%) had malignant neoplasia, which included: inflamed adenocarcinoma (n = 5); mast cell tumor (n = 1); or squamous cell carcinoma (n = 1). PCR testing on biopsy specimens from 24 cats revealed feline papillomavirus type 2 DNA in only four cats. Cystadenomatosis was more prevalent in senior non-purebred cats, over-represented in male cats and did not appear to be associated with papillomavirus, feline infectious peritonitis, feline immunodeficiency virus/feline leukemia virus status or other identifiable illnesses. Further studies are needed to investigate the causes of cystadenomatosis.

Identification of A Novel Papillomavirus Associated with Squamous Cell Carcinoma in A Domestic Cat.

Papillomaviruses infect the skin and mucosal surfaces of diverse animal hosts with consequences ranging from asymptomatic colonization to highly malignant epithelial cancers. Increasing evidence suggests a role for papillomaviruses in the most common cutaneous malignancy of domestic cats, squamous cell carcinoma (SCC). Using total DNA sequencing we identified a novel feline papillomavirus in a nasal biopsy taken from a cat presenting with both nasal cavity lymphoma and recurrent squamous cell carcinoma affecting the nasal planum. We designate this novel virus as Felis catus papillomavirus 6 (FcaPV6). The complete FcaPV6 7453 bp genome was similar to those of other feline papillomaviruses and phylogenetic analysis revealed that it was most closely related to FcaPV3, although was distinct enough to represent a new viral type. Classification of FcaPV6 in a new genus alongside FcaPVs 3, 4 and 5 is supported. Archived excisional biopsy of the SCC, taken 20 months prior to presentation, was intensely positive on p16 immunostaining. FcaPV6, amplified using virus-specific, but not consensus, PCR, was the only papillomavirus detected in DNA extracted from the SCC. Conversely, renal lymphoma, sampled at necropsy two months after presentation, tested negative on FcaPV6-specific PCR. In sum, using metagenomics we demonstrate the presence of a novel feline papillomavirus in association with cutaneous squamous cell carcinoma.

A virome sequencing approach to feline oral squamous cell carcinoma to evaluate viral causative factors

Feline oral squamous cell carcinoma (FOSCC) may be the best naturally-occurring model of human head and neck squamous cell carcinoma (HNSCC). HNSCC can be broadly divided into human papillomavirus (HPV)-negative cancers and HPV-positive cancers where HPV is the causative agent. Previous studies in FOSCC have used both species-specific and species-nonspecific PCR primers that may be insensitive to the detection of PVs and other viruses that may be divergent from known sequences. ViroCap is a targeted capture and next generation sequencing tool that was designed to identify all known vertebrate DNA and RNA viruses. In this study we used a metagenomic approach using ViroCap for DNA viruses in 20 FOSCC, 9 normal feline oral mucosal, and 8 suspected PV positive control samples. We tested the hypothesis that viruses would be enriched in FOSCC compared to normal oral mucosa. The virome of the FOSCC and normal feline oral mucosa consisted of feline foamy virus in 7/20 and 2/9 (35% and 22%), feline torque teno virus in 2/20 and 0/9 (10% and 0%), alphaherpesvirus in 2/10 and 0/9 (10% and 0%), FIV (0% and 22%), Epstein-Barr virus in 1/20 and 0/9 (5% and 0%) and feline papillomavirus in 1/20 and 0/9 samples (5% and 0% respectively). Felis catus papillomavirus-3 was found in 1 of 20 FOSCC samples. A virus was not associated consistently with FOSCC. If PVs have a role in FOSCC it is at most a supplementary or uncommon role. FOSCC appears most closely related to HPV-negative HNSCC. Future research on FOSCC should focus on identifying genetic and environmental causes.

Small Animal Review

Introduction: Productive feline papilloma virus infection is a cause of squamous cell carcinoma in UV-protected sites.

An Update on Canine, Feline and Bovine Papillomaviruses.

Over recent years, a growing number of papillomaviruses have been identified, which cause a wide range of lesions in domestic and wild animals. Papillomavirus-induced lesions may have a great impact on animal health, and some diseases observed in farm animals are associated with significant economic losses. This concise review brings together recent advancements on animal papillomavirus research, providing the scientific community and veterinary practitioners with an update on this rapidly evolving field. Among others, bovine, canine and feline papillomaviruses (BPV, CPV and FcaPV) are most extensively discussed, in view of the recent discovery of new viral types and their worldwide importance for animal health. Feline papillomaviruses 2 is an emerging, highly prevalent pathogen in domestic cats, associated with a subset of malignant skin lesions. Aspects related to cross-species infection by BPV and its environmental co-factors are also addressed. Animal papillomaviruses are also fascinating models for studying molecular and cell biology and have recently inspired some major breakthroughs. Overall, it is clear that additional, international and systematic efforts are needed to clarify which lesions are caused by which viral types and to develop experimental models for studying animal papillomavirus.

P16, pRb, and p53 in Feline Oral Squamous Cell Carcinoma.

Feline oral squamous cell carcinoma (FOSCC) is a highly aggressive head and neck cancer in cats, but the molecular pathogenesis of this cancer is still uncertain. In this study, p16, p53, and pRb proteins were detected and quantified by immunohistochemistry in forty-three FOSCC primary tumors and three FOSCC xenografts. p16 mRNA levels were also measured in three FOSCC cell lines (SCCF1, F2, and F3), which were consistent with their p16 immunoreactivity. Feline SCCF1 cells had very high levels of p16 protein and mRNA (55-fold greater) compared to SCCF2 and F3. A partial feline p16 cDNA sequence was amplified and sequenced. The average age of cats with FOSCC with high p16 immunoreactivity was significantly lower than the average age in the low p16 group. Eighteen of 43 (42%) FOSCCs had low p16 intensity, while 6/43 (14%) had high p16 immunoreactivity. Feline papillomavirus L1 (major capsid) DNA was not detected in the SCC cell lines or the FOSCCs with high p16 immunostaining. Five of 6 (83%) of the high p16 FOSCC had low p53, but only 1/6 (17%) had low pRb immunoreactivity. In summary, the staining pattern of p16, p53, and pRb in FOSCC was different from human head and neck squamous cell carcinoma and feline cutaneous squamous cell carcinoma. The majority of FOSCCs have low p16 immunostaining intensity, therefore, inactivation of CDKN2A is suspected to play a role in the pathogenesis of FOSCC. A subset of FOSCCs had increased p16 protein, which supports an alternate pathogenesis of cancer in these cats.

Feline Papillomavirusinfektion bei einer Katze mit Bowen-like disease und kutanem Plattenepithelkarzinom

A cutaneous mass in the neck was excised in a 13-year-old cat. Histopathological examination of the resected tissue revealed a multicentric squamous cell carcinoma in situ resembling Bowen's disease of man. The tumor showed a multifocal transformation to an infiltrative squamous cell carcinoma. Histological and immunohistological findings excluded actinic keratosis and feline viral plaques and allowed a classification as an irregular non-hyperkeratotic type of multicentric squamous cell carcinoma in situ. As a possible causative agent feline papillomavirus type 2 was detected using nested PCR in formalin-fixed material.

Molecular characterization of the L1 gene of papillomaviruses in epithelial lesions of cats and comparative analysis with corresponding gene sequences of human and feline papillomaviruses

To characterize the L1 gene of papillomaviruses detected in epithelial lesions of cats and to determine the relationship between those L1 gene nucleotide sequences and known L1 gene sequences of human and feline papillomaviruses. 10 tissue samples of epithelial lesions from 8 cats. DNA was extracted from tissue samples. Primers were designed to amplify the L1 gene of papillomaviruses. Amplicons of DNA were sequenced; nucleotide sequences were compared with known L1 gene nucleotide sequences of papillomaviruses and used for phylogenetic analysis. Tissue samples were obtained from lesions (diagnosed as dysplasia [n=1], squamous cell carcinoma in situ [3], or squamous cell carcinoma [6]) of the skin (9) and oral mucosa [1]. Two amplicons had 99% homology with the L1 gene nucleotide sequence of human papillomavirus type 38b subtype FA125. Another amplicon had 84% homology with the L1 gene nucleotide sequence of human papillomavirus type 80 and was considered to be a new type of papillomavirus. Phylogenetic tree analysis revealed that these 3 papillomaviruses were grouped into 2 clades that were not similar to the clades of Felis domesticus papillomavirus type 1 or F domesticus papillomavirus type 2 (FdPV2). The remaining 7 amplicons had 98% to 100% homology with the L1 gene nucleotide sequence of FdPV2. Phylogenetic tree analysis revealed that those 7 papillomaviruses were grouped nto a single clade with FdPV2. Results support the likelihood of transmission of papillomaviruses between humans and cats.

Detection of two different papillomaviruses within a feline cutaneous squamous cell carcinoma: Case report and review of the literature

CASE HISTORY A 15-year-old, brown-and-white cat was presented to a veterinary clinic with an ulcerated, reddened 1-cm diameter lesion on the nasal planum. CLINICAL FINDINGS AND DIAGNOSIS: Histology of a biopsy sample confirmed the lesion was a squamous cell carcinoma (SCC). PCR amplified DNA sequences from two different papillomaviruses. One sequence was from FdPV 2, which has previously been amplified from feline cutaneous SCC. However, the other sequence has not previously been reported, suggesting a novel feline papillomavirus. CLINICAL RELEVANCE: There is evidence that papillomaviruses promote the development of SCC on sun-exposed skin in humans. This is the first report in a cat of a papillomavirus other than FdPV2 and the first time that multiple papillomaviruses have been detected within a single neoplasm in this species. Whether the papillomaviruses influenced the development and behaviour of this SCC is currently uncertain, but this case provides additional evidence of the association between papillomaviruses and feline cutaneous SCC. If papillomaviruses are found to influence the development of SCC this may allow novel strategies to prevent these common neoplasms in cats.

Ancient papillomavirus-host co-speciation in Felidae

The evolutionary rate of feline papillomaviruses is inferred from the phylogenetic analysis of their hosts, providing evidence for long-term virus-host co-speciation

Detection of novel papillomaviruslike sequences in paraffin-embedded specimens of invasive and in situ squamous cell carcinomas from cats

To detect and partially characterize papillomavirus (PV) DNA in squamous cell carcinoma (SCC) tumor specimens from cats. 54 formalin-fixed paraffinembedded skin biopsy specimens were examined. Specimens originated from Bowenoid in situ SCC (BISC; n = 21), invasive SCC (22), and skin affected by miscellaneous nonneoplastic conditions (11). Samples from each tissue block underwent DNA extraction after deparaffinization, and PCR assays were performed. Two sets of primers derived from PV E1 were used. The first set of primers was designed for the narrow-range PCR assay and was able to generate amplification products of feline PV (FePV), canine oral PV, or closely related PVs. The second set of primers was selected for the broad-range PCR assay because of its ability to amplify DNA from 64 human PVs. Sequence analysis of each amplified DNA was performed. 1 of the 21 specimens of BISC was positive for PV DNA on the basis of narrow-range PCR assay results, whereas all the other specimens (BISC, invasive SCC, and controls) had negative results for PV DNA. In contrast, 5 of 21 BISC specimens and 4 of 22 invasive SCC specimens were positive for PV DNA on the basis of broad-range PCR assay results. Sequence analysis revealed that only 1 specimen was infected by a virus closely related to classic FePV. In the 8 other specimens positive for PV DNA, DNA of unknown PVs was uncovered. Bowenoid in situ SCC and invasive SCC of cats may be associated with PVs of genetic diversity.

Detection of novel papillomaviruses in canine mucosal, cutaneous and in situ squamous cell carcinomas

Papillomavirus (PV) DNA is frequently uncovered in samples of human skin squamous cell carcinomas (SCC). However, the role of these viruses in the development of such cancers in canine species remains controversial. While approximately 100 human PVs are known, only one single canine oral PV (COPV) has been identified and studied extensively. Therefore, we applied a narrow-range polymerase chain reaction (PCR) suitable for the detection of classical canine and feline PVs, as well as a broad-range PCR, which has been used for the detection of various novel PVs in humans, in order to analyse 42 paraffin-embedded samples, representing three different forms of canine SCCs. Ten samples of skin tissues with various non-neoplastic conditions served as controls. While none of the negative controls reacted positively, PV DNA was discovered in 21% of the tested SCC samples. Interestingly, the classical COPV was amplified from only one sample, while the other positive cases were associated with a variety of thus far unknown PVs. This study suggests that a fraction of canine SCC is infected with PVs and that a genetic variety of canine PVs exists. Therefore, these results will facilitate the future study of the role of PVs in the development of canine skin cancers.