M. S. Campo

Bovine papillomavirus and cancer

Bovine papillomavirus (BPV) induces papillomas of cutaneous or mucosal epithelia in cattle. The papillomas are benign tumours and generally regress, but occasionally persist and provide the focus for malignant transformation to squamous cell carcinoma, particularly in the presence of environmental cofactors. This has been experimentally demonstrated for BPV-2 and cancer of the urinary bladder, and BPV-4 and cancer of the upper alimentary canal in cattle feeding on bracken fern. In this review, several aspects of the biology of the virus are described including viral genome structure, regulation of transcription of the viral oncogenes, function of the viral oncoproteins, cooperation between virus and chemical cofactors in carcinogenesis, virus latency and prophylactic and therapeutic vaccination programmes.

Prophylactic and therapeutic vaccination against a mucosal papillomavirus

Papillomaviruses are ubiquitous DNA viruses affecting humans and animals and causing a variety of tumours of mucosal and cutaneous epithelia. Some of these lesions, particularly those affecting mucosal epithelia, can progress to squamous cell carcinomas. Prevention or cure of viral infection would ultimately lead to a decrease in the incidence of papillomavirus-associated cancers. Using recombinant proteins, we have developed prophylactic and therapeutic vaccines against bovine papillomavirus type 4, a mucosal papillomavirus implicated in cancer of the alimentary canal in cattle; similar possibilities exist for the human mucosal papillomaviruses.

Papillomavirus E5: the smallest oncoprotein with many functions

Papillomaviruses (PVs) are established agents of human and animal cancers. They infect cutaneous and mucous epithelia. High Risk (HR) Human PVs (HPVs) are consistently associated with cancer of the uterine cervix, but are also involved in the etiopathogenesis of other cancer types. The early oncoproteins of PVs: E5, E6 and E7 are known to contribute to tumour progression. While the oncogenic activities of E6 and E7 are well characterised, the role of E5 is still rather nebulous. The widespread causal association of PVs with cancer makes their study worthwhile not only in humans but also in animal model systems. The Bovine PV (BPV) system has been the most useful animal model in understanding the oncogenic potential of PVs due to the pivotal role of its E5 oncoprotein in cell transformation. This review will highlight the differences between HPV-16 E5 (16E5) and E5 from other PVs, primarily from BPV. It will discuss the targeting of E5 as a possible therapeutic agent.

Studies on vaccination against papillomaviruses: Prophylactic and therapeutic vaccination with recombinant structural proteins

The L1 and L2 proteins of BPV-2 have been produced in Escherichia coli as beta-galactosidase fusion proteins. The fusion proteins have been used to vaccinate calves both prophylactically and therapeutically. The L1 fusion protein prevented tumor formation when administered before challenge with BPV-2, while the L2 fusion protein was very effective in promoting tumor rejection, independently from whether it was administered before or after challenge. Animals vaccinated with L1, but not with L2, responded rapidly with production of serum neutralizing antibodies, showing that this peptide contains B-cell-specific epitopes. The massive infiltration of lymphocytes in the tumors of L2-vaccinated animals suggests that the peptide contains epitopes specific for T-cells. The two structural proteins of BPV-2 therefore interact with both efferent arms of the immune system, and this observation allows the choice between two different types of antiviral vaccination.

Latent papillomavirus infection in cattle

During a long term experiment designed to identify the contribution of bovine papillomavirus type 4 (BPV-4), environmental mutagens and immunosuppressants to the development of carcinomas of the upper alimentary tract of cattle, there was evidence of latent papillomavirus infection. Papillomatosis-free animals, when immunosuppressed either by feeding bracken fern or by azathioprine treatment, developed skin warts containing either BPV-1 or BPV-2. Skin warts appeared also in an immunocompetent animal at sites of damaged skin. It was concluded that the animals harboured latent papillomavirus which was reactivated by immunosuppression and/or physical trauma, causing skin warts. Papillomavirus DNA was also detected in lymphocytes of both experimental and control animals, suggesting that one of the sites of latency may be the circulating lymphocyte.

Human papillomavirus DNA in biopsies of oral tissues

The DNAs of human papillomavirus (HPV) types 4, 16 and 18 have been detected in biopsies of normal and malignant human oral mucosa by Southern blot hybridization and the polymerase chain reaction (PCR). By the former technique, HPV-4, HPV-16 and HPV-18 DNAs were detected in three separate carcinomas but were found in adjacent dysplastic and normal tissue by the PCR only. The PCR technique also allowed detection of HPV-16 and HPV-18 DNA in additional carcinomas and normal samples. The oral HPV-4 DNA was molecularly cloned and extensive restriction analysis and nucleotide sequencing showed identity with the prototype HPV-4 DNA. The HPV-18 DNA detected by Southern blot hybridization showed an altered restriction pattern in the E1 region of the viral genome; however direct nucleotide sequencing of PCR products from the E6 open reading frame showed no sequence alterations in either normal or malignant samples.

Sequence variants of bovine papillomavirus E5 detected in equine sarcoids

The equine sarcoid, one of the most common dermatological lesions in equids, is a benign, locally invasive dermal fibroblastic lesion. Previous studies have suggested an association with two bovine papilloma virus (BPV) types, BPV-1 and BPV-2. In the present study, we examined sarcoids from horses from two geographical areas, Switzerland and the UK, for the major transforming gene of BPV, E5. We detected BPV DNA for the E5 open reading frame and viral E5 RNA transcripts in most sarcoids. Sequence analysis of the E5 open reading frame of sarcoid-associated BPV detected several unique DNA sequence variants, three of which resulted in sarcoid specific amino acid sequence variations. It is unclear if these sequence variants contribute to the unique clinical presentation of the sarcoid. However, our work provides further evidence of the association between BPV and sarcoid development and the direct involvement of the virus in the pathogenesis of sarcoids.

HPV-16 E5 down-regulates expression of surface HLA class I and reduces recognition by CD8 T cells

HPV-16 is the major causes of cervical cancer. Persistence of infection is a necessary event for progression of the infection to cancer. Among other factors, virus persistence is due the viral proteins fighting the immune response. HPV-16 E5 down-regulates MHC/HLA class I, which is much reduced on the cell surface and accumulates in the Golgi apparatus in cells expressing E5. This effect is observed also in W12 cells, which mimic early cervical intraepithelial progression to cervical cancer. The functional effect of MHC I down-regulation on human CD8 T cells is not known, because of the need for HLA-matched, HPV-specific T cells that recognise E5 expressing-cells. Here we employ a heterologous cell/MHC I system which uses mouse cells expressing both E5 and HLA-A2, and A2-restricted CTLs; we show that the E5-induced reduction of HLA-A2 has a functional impact by reducing recognition of E5 expressing cells by HPV specific CD8+ T cells.

Vaccination of cattle with bovine papillomavirus type 4 L2 elicits the production of virus-neutralizing antibodies

Prophylactic vaccination of cattle with the N terminus (L2a, aa 11-200) of the minor capsid protein L2 completely prevented bovine papillomavirus type 4 (BPV-4) infection of the alimentary canal. To investigate the mechanisms underlying protection from viral infection, sera from vaccinated animals were analysed in neutralization assays both in the nude mouse xenograft system and in cattle. BPV-4 retained its infectivity when incubated with preimmune cattle sera, whereas, when incubated with immune sera from animals vaccinated with either whole L2 or its N terminus L2a, its infectivity was greatly reduced, indicating that the immune sera had neutralizing activity against the virus. This activity could be abrogated by absorbing the immune sera with L2 or L2a, thus indicating that virus neutralization was due to the presence in the immune sera of anti-L2 antibodies.

A novel bovine papillomavirus (BPV-6) causing true epithelial papillomas of the mammary gland skin: a member of a proposed new BPV subgroup

A papillomavirus has been isolated from frond epithelial papillomas of the bovine udder. It is clearly distinguishable from all other bovine papillomaviruses (BPVs) based on DNA sequence homology and antigenic properties and is thus characterised as a new entity, designated BPV-6. BPV-6 does not possess the interspecific papillomavirus antigen, its genomic DNA (7.2 kb) is smaller than, and does not show any sequence homology to BPV-1, BPV-2, or BPV-5, whereas it is approximately the same length as BPV-3 or BPV-4, with which it shares some sequence homology. The bovine papillomaviruses have been classified into two subgroups: subgroup A, composed of BPV-1, BPV-2, and BPV-5, all of which induce fibropapillomas, and subgroup B, composed of BPV-3, BPV-4, and BPV-6, all of which cause true epithelial papillomas.