Neroli A. Thomson

The use of quantitative PCR to detect Felis catus papillomavirus type 2 DNA from a high proportion of queens and their kittens

Squamous cell carcinomas are common feline skin cancers that have been associated with infection with Felis catus papillomavirus type 2 (FcaPV-2). Currently, little is known about the epidemiology of FcaPV-2 infection. The aim of this study was to develop a real-time PCR assay to quantify FcaPV-2 DNA in plucked hairs and skin swabs from 11 healthy breeding queens and their kittens. Samples were taken prior to kittening and then 2, 7 and 28 days after kittening to determine the age at which the kittens were first exposed to the virus. FcaPV-2 DNA was amplified from all of the queens and from 91% of the kittens at 2 days of age. There was a wide range in the quantity of FcaPV-2 DNA detected, from 1 to 92,520 copies per swab, and from 0.01 to 234 copies per copy of reference gene DNA in the hair plucks. The quantity of FcaPV-2 DNA detected in samples collected from the kittens was strongly correlated to that of their respective queens and the mean viral DNA load was similar for cats within a household but varied significantly between households. This is the first time that quantitative PCR has been used to detect FcaPV-2 DNA and the results suggest that the virus is ubiquitous but there is a wide variation of viral DNA loads. Kittens appear to be exposed to FcaPV-2 early in life, presumably from direct contact with their queen. These results are important when determining if FcaPV-2 infection of cats is preventable.

Papillomaviruses in dogs and cats

Papillomaviruses (PVs) cause disease in both dogs and cats. In dogs, PVs are thought to cause oral papillomatosis, cutaneous papillomas and canine viral pigmented plaques, whereas PVs have been rarely associated with the development of oral and cutaneous squamous cell carcinomas in this species. In cats, PVs are currently thought to cause oral papillomas, feline viral plaques, Bowenoid in situ carcinomas and feline sarcoids. Furthermore, there is increasing evidence that PVs may also be a cause of cutaneous squamous cell carcinomas and basal cell carcinomas in cats. These diseases are discussed in this review. Additionally, there is a brief overview of PV biology, including how these viruses cause disease. Diagnostic techniques and possible methods to prevent PV infection are also discussed.

Detection of DNA sequences from a novel papillomavirus in a feline basal cell carcinoma

BACKGROUND Basal cell carcinomas (BCCs) are uncommon feline skin neoplasms of uncertain cause. CASE A 14-year-old Abyssinian cat developed a soft dermal nodule on the dorsal thorax. This mass grew slowly over a six month period before being surgically excised. METHODS AND RESULTS Histology revealed a BCC. Additionally, changes suggestive of an early Bowenoid in situ carcinoma (BISC) were present in the overlying epidermis. Both the BCC and the BISC contained papillomavirus-induced cell changes and prominent basophilic intracytoplasmic bodies. PCR using consensus primers and primers specific for Felis catus papillomavirus types 2 and 3 (FcaPV-2 and -3) was used to amplify papillomaviral DNA. The same papillomaviral DNA sequence was present in the BCC and the BISC. This sequence was most similar to FcaPV-3, but with just 70.5% similarity, was from a novel papillomavirus type. No recurrence or further masses developed. CONCLUSIONS This case is unusual due to the presence of a large dermal BCC associated with minimal BISC changes in the overlying epidermis. Additionally, papillomavirus-induced cell changes have not been described previously in a BCC. Furthermore, both the BCC and the BISC contained sequences from a novel papillomavirus type. These observations suggest that the development of some BCCs could be influenced by papillomavirus infection. The novel papillomavirus type detected is the third papillomavirus type to be associated with skin cancer in cats.

Bilateral pre-auricular papillary squamous cell carcinomas associated with papillomavirus infection in a domestic cat

BACKGROUND Cutaneous papillary squamous cell carcinomas (SCCs) are extremely rare in humans and have not been reported in any nonhuman species. In humans, oral papillary SCCs are often caused by papillomavirus infection and have a more favourable prognosis than other SCC subtypes. CASE A 10-year-old ginger and white domestic short hair cat had a 12 month history of symmetrical, roughly circular, exophytic 2 cm diameter masses in both pre-auricular regions. Surgical excision was performed, although with only narrow margins. METHODS AND RESULTS Histology of both masses revealed a proliferation of neoplastic keratinocytes arranged in numerous filiform projections that were supported by fibrovascular stalks. Although the cells were confined to the epidermis predominantly, nests of neoplastic cells were visible within the superficial dermis. The neoplastic cells demonstrated significant atypia with a variable nuclear:cytoplasmic ratio and a high mitotic index. A papillary subtype SCC was diagnosed. Felis catus papillomavirus type 2 (FcaPV-2) was the only papillomavirus detected in the masses and FcaPV-2 E6/E7 gene expression and p16CDKN2A protein immunostaining were detected. Six months after surgery neither recurrence nor further masses had developed. CONCLUSIONS This is the first cutaneous papillary SCC reported in a nonhuman species. Papillary SCCs may be a rare manifestation of FcaPV-2 infection in cats. The unusual location of the SCCs suggests that both papillomavirus infection and ultraviolet light exposure could have contributed to neoplasia development. Evidence from this single case suggests that papillary SCCs may have a more favourable prognosis than conventional SCCs in cats.

Multiple viral plaques with sebaceous differentiation associated with an unclassified papillomavirus type in a cat

Abstract CASE HISTORY AND CLINICAL FINDINGS A 15-year-old neutered male domestic short-haired cat was presented due to multiple 0.5–2 cm-diameter crusting plaques in the left preauricular region, over the bridge of nose, and in the right periocular region. The plaques did not appear to cause discomfort. HISTOPATHOLOGICAL FINDINGS Biopsy samples of four plaques were examined histologically. Three plaques consisted of well-demarcated foci of mild epidermal hyperplasia overlying markedly hyperplastic sebaceous glands. Approximately 60% of the hyperplastic cells contained a large cytoplasmic vacuole that ranged from being clear to containing prominent grey-blue fibrillar material. The fourth plaque was composed solely of epidermal hyperplasia, consistent with previous descriptions of feline viral plaques. MOLECULAR BIOLOGY Papillomavirus DNA was amplified from all four plaques using PCR. A single DNA sequence was amplified from the plaques with sebaceous differentiation. This sequence was identical to the FdPV-MY sequence previously suggested to be from a putative unclassified papillomavirus type. Felis catus papillomavirus type 2 sequences were amplified from the plaque typical of feline viral plaques. Immunohistochemistry to detect p16CDKN2A protein (p16) showed marked immunostaining throughout the hyperplastic epidermis and adnexal structures within the plaques with sebaceous differentiation. DIAGNOSIS Multiple feline viral plaques with variable sebaceous differentiation. CLINICAL RELEVANCE Feline viral plaques with sebaceous differentiation have not been previously reported in cats. The presence of unique cell changes within these lesions, the detection of an unclassified papillomavirus type, and the p16 immunostaining within these plaques suggest that they may have been caused by the papillomavirus that contains the FdPV-MY sequence.

Anal fibropapillomas containing bovine papillomavirus type 2 DNA in two groups of heifers

Abstract CASE HISTORY Anal warts were observed in heifers in two unrelated groups of animals. Heifers in one group developed visible warts 4 months after manual rectal examination and heifers in the other group developed warts 5 months after examination using a hand-held rectal ultrasound probe. CLINICAL FINDINGS Large exophytic proliferative anal masses were observed in 5/15 (33%) heifers in one group and 13/149 (9%) heifers in the second group. Heifers in the second group were also noted to have similar masses on the underside of the tail at sites previously used for venepuncture and some of the heifers had skin warts. Despite the large size of the anal masses, none of the heifers showed clinical signs of systemic illness. HISTOPATHOLOGICAL FINDINGS An anal mass was removed from one heifer in each of the two groups. Sections from both masses showed hyperplastic epithelium covering a proliferation of well-differentiated fibroblasts consistent with fibropapillomas. Small numbers of cells within the epidermis had clear cytoplasm with clumped keratohyalin granules. MOLECULAR BIOLOGY Bovine papillomavirus (BPV) type 2 DNA was amplified from both fibropapillomas by PCR. DIAGNOSIS Multiple anal fibropapillomas associated with BPV-2. CLINICAL RELEVANCE Bovine anal fibropapillomas have only been reported in heifers that have undergone rectal examination, and infection of anal microabrasions in an immunologically naïve animal appears to be associated with disease development. The source and method of spread of BPV-2 within these groups could not be determined. However spread of BPV-2 within the groups by the veterinarian performing rectal examinations may have been most likely. While these fibropapillomas had a dramatic appearance, like fibropapillomas elsewhere on the body, they did not have any significant effect on the health of the affected heifers. As these lesions can be diagnosed by clinical examination and self-resolve without treatment, it is important that veterinarians are aware of this rare manifestation of papillomavirus infection of cattle.

Identification of Felis catus papillomavirus 3 in skin neoplasms from four cats

Bowenoid in situ carcinomas (BISCs) are papillomavirus (PV)-induced skin neoplasms that are thought to be caused by Felis catus papillomavirus (FcaPV) 2. As BISCs are typically multiple and can become extensive, they can be difficult to treat. Herein we describe 4 cats that developed skin neoplasms that contained FcaPV-3 DNA. One cat developed multiple basal cell carcinomas (BCCs), 1 a BISC with unusual extension into hair follicles, and 2 developed a single typical-appearing BISC. All neoplasms contained prominent PV-induced cell changes and intense p16CDKN2a protein immunostaining. Results from these 4 cats provide evidence that FcaPV-3 could cause a proportion of feline skin cancers, albeit less frequently than FcaPV-2. Excision of the typical BISCs and the BCCs appeared curative. Although the cat with the unusual BISC was euthanized because of the large size of the lesion, evidence from these 4 cats suggests that skin neoplasms that contain FcaPV-3 DNA may have a less aggressive clinical behavior than those associated with FcaPV-2. A consistent feature of the neoplasms in all 4 cats was the presence of prominent basophilic intracytoplasmic inclusion bodies; these inclusions have not been reported in lesions caused by FcaPV-2, to our knowledge, and their detection may allow differentiation between the different PV types and could therefore be a useful prognostic feature.

Felis catus papillomavirus type 2 DNA loads on kittens are transient and do not reflect their susceptibility to infection

Objectives Felis catus papillomavirus type 2 (FcaPV-2) commonly infects the skin of domestic cats, and mounting evidence suggests that the virus could be involved in a subset of feline skin cancers. The reason why some cats develop FcaPV-2-induced disease and others do not is currently unknown. However, it has been shown that kittens in different litters have markedly different FcaPV-2 DNA loads and the aim of this study was to determine whether these differences could be due to inherent differences in susceptibility to infection. Such differences could potentially explain why only a small proportion of cats develop FcaPV-2-associated skin disease. Methods Repeated skin swabs were taken to measure FcaPV-2 DNA loads in queens and kittens in a research colony. The kittens either stayed in their original litters or were moved between litters; eventually, all of the kittens were housed together. A subset of samples was also analysed for FcaPV-2 mRNA. Results While there were initially large differences in FcaPV-2 DNA loads between litters of kittens, these differences disappeared when the kittens were moved between litters or housed together. Importantly, the viral DNA loads changed too rapidly to be due to the acquisition or clearance of infection. In contrast, the differences in viral DNA loads between the different queens were sustained throughout the experiment. FcaPV-2 mRNA was also detected in samples from 1- to 8-day-old kittens. Conclusions and relevance The results suggest that the FcaPV-2 DNA load in a swab sample from an individual kitten largely reflects the overall level of FcaPV-2 shedding in the group of in-contact cats, rather than the infection status of the individual kitten. Therefore, there was no evidence for inherent differences in susceptibility to infection. However, the finding of FcaPV-2 mRNA suggests that at least some kittens do become infected with FcaPV-2 early in life.