Papillomaviruses: a systematic review

Abstract In the last decades, a group of viruses has received great attention due to its relationship with cancer development and its wide distribution throughout the vertebrates: the papillomaviruses. In this article, we aim to review some of the most relevant reports concerning the use of bovines as an experimental model for studies related to papillomaviruses. Moreover, the obtained data contributes to the development of strategies against the clinical consequences of bovine papillomaviruses (BPV) that have led to drastic hazards to the herds. To overcome the problem, the vaccines that we have been developing involve recombinant DNA technology, aiming at prophylactic and therapeutic procedures. It is important to point out that these strategies can be used as models for innovative procedures against HPV, as this virus is the main causal agent of cervical cancer, the second most fatal cancer in women.

Caracterização molecular de DNA de Delta papillomavirus bovino (BPV1, 2 e 13) em sarcoides equinos / Molecular characterization of bovine Deltapapillomavirus (BPV1, 2, and 13) DNA in equine sarcoids

Sarcoides são tumores fibroblásticos, considerados os tumores de pele mais comuns em pele de equinos e que raramente apresentam regressão espontânea. Papilomavírus bovino (BPV) tipos 1 e 2 são relacionados com a patogenia do sarcoide e, provavelmente, o BPV tipo 13 (BPV13), recentemente descrito, também pode estar associado com a formação dessa lesão. Neste estudo, 20 amostras de lesões cutâneas, sendo 12 constituídas por tecidos frescos e 8 amostras de tecido fixado em formalina e embebido em parafina, provenientes de 15 cavalos foram utilizadas para a identificação do DNA de BPV. A análise histopatológica (HE) confirmou todas as lesões como sarcoide. Para a amplificação do DNA de papilomavírus (PV) foram realizadas três reações de PCR. Como triagem, os primers IFNR2/IDNT2 foram utilizados para amplificar um fragmento da ORF L1 do PV. O segundo par de primersutilizado é complementar a sequência dos genes E5 e L2 de BPVs 1, 2 e 13. O terceiro par de primers(FAP59/FAP64) utilizado tem o gene L1 como alvo. A primeira e a segunda PCRs permitiram amplificar produtos em todas as amostras avaliadas. Entretanto, na terceira reação, na qual foram utilizados os primers FAP, foi possível amplificar produtos com tamanho molecular esperado somente nas amostras constituídas por tecidos frescos. O sequenciamento de nucleotídeos e as análises filogenéticas realizadas nos fragmentos E5L2 resultaram na identificação de BPV1, 2 e 13 em 14 (70%), 2 (10%) e em 4 (20%) amostras de sarcoides, respectivamente. As amostras de sarcoides de um dos animais continha somente o DNA de BPV1. Entretanto, nas amostras provenientes do segundo cavalo foi possível identificar o DNA de três tipos de Deltapapillomavirus bovino (BPV1, 2 e 13) em lesões distintas. Este estudo ratifica a presença do DNA de BPV1, 2 e 13 em lesões de sarcoides em equinos, além de identificar três tipos de BPVs em um mesmo animal e descrever pela primeira vez no Brasil a presença de BPV1 e 2 nesse tipo de lesão.

Sarcoids are fibroblastic lesions, which are considered as the most common skin tumors of horses; spontaneous regression rarely occurs. The bovine papillomavirus (BPV) types 1 and 2 may be involved in the pathogenesis of sarcoids, and probably the recently described BPV type (BPV13) might be associated with the pathogenesis of this lesion. This study characterized the DNA of BPVs in sarcoids from 15 horses from Brazil by analyzing 20 cutaneous lesions (12 recently collected; 8 from formalin-fixed paraffin-embedded (FFPE) tissues). Histopathology confirmed the proliferative lesions as sarcoids. Three PCRs were performed to amplify papillomavirus (PV) DNA. For screening, the primers IFNR2/IDNT2 were used to amplify a fragment of the PV L1 ORF. The second primer set was complementary to a common sequence of the E5L2 genomic region of BPV1, 2, and 13. The third primer pair (FAP59/FAP64) targeted a fragment of the PVs L1 ORF. The screening and E5L2 PCRs yielded amplicons in all samples evaluated. The FAP amplicons identified BPV1, 2, and 13 only from fresh tissue samples. The phylogenetic analyses of E5L2 resulted in the identification of BPV1, 2, and 13 in 14 (70%), 2 (10%), and 4 (20%) sarcoids, respectively. Two horses demonstrated multiple lesions: the sarcoids of one of these contained only BPV1 DNA and those of the other contained three types of bovine Deltapapillomavirus (BPV1, 2, and 13). This study confirmed the presence of BPV1, 2, and 13 DNA in equine sarcoids. Moreover, these findings represent the first description of three types of BPV diagnosed in the same horse, as well as the first confirmation of BPV1 and 2 in horses from Brazil.

Teat papillomatosis associated with bovine papillomavirus types 6, 7, 9, and 10 in dairy cattle from Brazil

This study describes the clinical, histopathological, and virological characterization of teat papillomatosis from Brazilian dairy cattle herds. Four types of bovine papillomavirus were identified (BPV6, 7, 9, and 10); one of these (BPV7) is being detected for the first time in Brazilian cattle.

Short term results of balloon pulmonary valvuloplasty in children: a single centre study over a period of 42 months.

Objectives: To evaluate the efficacy and safety of balloon pulmonary valvuloplasty (BPV) in children. Design, setting and method: This is a retrospective review of 101 consecutive patients with pulmonary stenosis who underwent BPV at the Lady Ridgeway Hospital for Children over a period of 42 months. Four patients were excluded as they had complex cardiac problems. Data was obtained from Excel based cardiac catheterization reports in the Department of Paediatric Cardiology. Reduction of pressure gradient across pulmonary valve to less than 25 mmHg was defined as a successful procedure. Results: Ninety seven BPVs were performed in 94 patients. The mean age, bodyweight and height were 19 months, 9.5 kg and 80 cm respectively. Sex distribution was equal. The mean of the balloon to pulmonary annular ratio was 108% (range 50-150%). After the BPV, mean peak to peak pressure gradient across the pulmonary valve dropped from 70±29 mmHg to 24.7±16.8 mmHg. Overall success rate was 55.7%. Mortality was 3.1%. Major complications were haemopericardium (4.1%), bradycardia (2%) endocardial damage (1%) and pulmonary oedema (1%). The correlation between pulmonary annulus on echocardiography and angiography was significant (p<0.01).

Phylogenetic position of an uncharacterized Brazilian strain of bovine papillomavirus in the genus Xipapillomavirus based on sequencing of the L1 open reading frame

The use of PCR assays with degenerate primers has suggested the existence of numerous as yet uncharacterized bovine papillomaviruses (BPV). Despite the endemic nature of BPV infections, the identification of BPV types in Brazilian cattle is still only sporadic. However, in a recent analysis of a partial segment of the L1 gene, we observed notable diversity among the BPV types detected. The aim of this study was to determine the phylogenetic position of the previously identified wild strain BPV/BR-UEL2 detected in the state of Paraná in Brazil. Since previous analysis of the partial L1 sequence had shown that this strain was most closely related to BPV type 4, genus-specific primers were designed. Phylogenetic analysis using complete L1 ORF sequences revealed that BPV/BR-UEL2 was related to BPV types classified in the genus Xipapillomavirus and shared the highest L1 nucleotide sequence similarity with BPV type 4 (78 percent). This finding suggests that BPV/BR-UEL2 should be classified as a potential new type of BPV in the genus Xipapillomavirus.

A bovine teat papilloma specimen harboring Deltapapillomavirus (BPV-1) and Xipapillomavirus (BPV-6) representatives

The common occurrence of multiple papillomavirus infections has been shown in several studies involving the human host. However, investigations with the aim of identifying mixed papillomavirus infections in cattle have been conducted only recently. In the current work we describe a co-infection with two different bovine papillomavirus (BPV) types that was identified in a bovine teat papilloma. The skin wart was obtained from a cow belonging to a Brazilian beef herd. A PCR assay was carried out with the FAP primer pair, which amplifies a partial segment of the L1 gene (approximately 478 bp), and the amplicon was submitted to direct sequencing. Because nucleotide sequences with satisfactory quality scores were not obtained, the amplicon was cloned and further sequencing, involving ten selected clones, was performed. The sequence analysis of the cloned inserts revealed the presence of two different BPV types. BPV-1 (Deltapapillomavirus genus) was detected in six clones, while BPV-6 (Xipapillomavirus genus) was detected in four clones. This finding confirms the presence of BPV co-infection associated with cutaneous papillomatosis in cattle.

Em seres humanos, as infecções múltiplas pelo papilomavírus têm sido demonstradas em vários estudos. Em bovinos, somente recentemente foram conduzidas investigações com o objetivo de avaliar infecções mistas pelo papilomavírus. O presente trabalho teve como objetivo descrever a co-infecção por dois tipos de papilomavírus bovino (BPV) em um papiloma de teto. A amostra clínica foi obtida de uma vaca pertencente a um rebanho de corte localizado na região norte do estado do Paraná, Brasil. Inicialmente, a técnica de PCR foi realizada com o par de oligonucleotídeos iniciadores FAP, que amplificam um segmento do gene L1, sendo que o amplicon gerado foi submetido ao sequenciamento direto. Entretanto, como as sequências obtidas não apresentaram qualidade aceitável, o amplicon foi clonado e dez clones foram selecionados para um novo sequenciamento. A análise das sequências dos insertos revelou a presença de dois diferentes tipos de BPV. O BPV-1 (gênero Deltapapillomavirus) foi detectado em seis clones, enquanto o BPV-6 (gênero Xipapillomavirus) foi detectado em quatro clones. Esse resultado confirma a ocorrência da co-infecção pelo BPV associada a papilomas cutâneos em bovinos.

Multiple bovine papillomavirus infections associated with cutaneous papillomatosis in brazilian cattle herds

Cutaneous papillomatosis is a pathological condition commonly found in cattle and is characterized by the presence of benign proliferative tumors caused by bovine papillomavirus (BPV) infection. While multiple infections with human papillomavirus (HPV) are common in healthy and immunodeficient humans, studies with the aim of identifying mixed infections are still sporadic in veterinary medicine. The aim of this study is to describe the occurrence of multiple BPV infections in cattle affected by cutaneous papillomatosis. Fifteen skin warts were collected from at least two diverse anatomical regions of six bovines with papillomatosis belonging to three cattle herds from the Paraná state in Brazil. The BPV types present in the skin wart samples were determined by a PCR assay performed with the FAP primer pair for partial L1 gene amplification followed by direct sequencing or by cloning and sequencing of the inserts. Sequence analysis of the obtained amplicons allowed the identification of four characterized BPV types (BPV-1, -2, -6, and -8) and three previously described putative new BPV types (BPV/BR-UEL3, BPV/BR-UEL4, and BPV/BR-UEL5). Double infections were identified in four (A, B, D, and E) of the six animals included in this study. In this work, the strategy adopted to evaluate skin warts from diverse anatomical sites of the same animal allowed the identification of multiple infections with two or three different BPV types. The analysis of four animals belonging to a single cattle herd also showed the presence of six different viral types. These results clearly suggest that both multiple papillomaviral infection and a high viral diversity can be as frequent in cattle as in human beings.

A papilomatose cutânea é comumente observada nos rebanhos bovinos e caracterizada pela presença de tumores proliferativos benignos causados pela infecção pelo papilomavírus bovino (BPV). Enquanto a infecção múltipla pelo papilomavírus humano (HPV) é um achado comum tanto em seres humanos saudáveis quanto em pacientes com imunodeficiência, na medicina veterinária esses relatos ainda são escassos. O objetivo desse estudo foi descrever a ocorrência de infecções múltiplas pelo BPV em rebanhos afetados pela papilomatose cutânea. Quinze papilomas foram obtidos, de pelo menos duas regiões anatômicas diferentes, de seis bovinos com papilomatose e provenientes de três rebanhos de corte localizados no estado do Paraná, Brasil. Os tipos virais presentes nas lesões foram identificados por PCR, utilizando o par de oligonucleotídeos iniciadores FAP, seguidos de sequenciamento direto ou clonagem e novo sequenciamento dos insertos. A análise das sequências obtidas permitiu a identificação do BPV-1, -2, -6 e -8, além de supostos novos tipos (BPV/BR-UEL3, BPV/BR-UEL4, e BPV/BR-UEL5), descritos anteriormente. Infecções por dois tipos diferentes de BPV foram identificadas em quatro animais (A, B, D e E) dos seis incluídos nesse estudo. A estratégia adotada neste estudo permitiu a identificação de infecção múltipla por dois ou três diferentes tipos virais do BPV no mesmo animal. Além disso, a avaliação de quatro animais de um mesmo rebanho demonstrou a presença de seis tipos virais circulantes. Esses resultados sugerem que tanto as infecções múltiplas quanto a grande diversidade viral podem ser frequentes nos bovinos, assim como o observado nos humanos. O reconhecimento da multiplicidade e complexidade das infecções pelo BPV pode colaborar para o entendimento dos aspectos epidemiológicos, clínicos e imunológicos da papilomatose cutânea nos rebanhos bovinos.

Identification of the recently described new type of bovine papillomavirus (BPV-8) in a Brazilian beef cattle herd: [revision] / Identificação do novo tipo de papilomavírus bovino (BPV-8) recentemente descrito num rebanho bovino no Brasil: [revisão]

Bovine papillomavirus type 8 (BPV-8) was first detected and described in teat warts as well as in healthy teat skin from cattle raised in Japan. The entire viral genome was sequenced in 2007. Additionally, a variant of BPV-8, BPV-8-EB, was also identified from papillomatous lesions of a European bison in Slovakia. In Brazil, despite the relatively common occurrence of BPV infections, the identification and determination of viral types present in cattle is still sporadic. The aim of this study is to report the occurrence of the recently described BPV-8 in Brazil. The virus was identified in a skin warts obtained from a beef cattle herd located in Parana state, southern Brazil. The papilloma had a macular, non-verrucous gross aspect and was located on the dorsal thorax of a cow. Polymerase chain reaction (PCR) was performed using generic primers for partial amplification of L1 gene. The obtained amplicon (480bp) was cloned and two selected clones were sequenced. The nucleotide sequence was compared to existing papillomaviral genomic sequences, identifying the virus as BPV type 8. This study represents the first report of BPV-8 occurrence in Brazil, what suggests its presence among Brazilian cattle.

A primeira descrição do papilomavírus bovino tipo 8 (BPV-8) foi realizada em amostras de papilomas de teto e de pele saudável de tetos de bovinos no Japão. Em 2007, a seqüência genômica completa do BPV-8 foi determinada. Ainda em 2007, uma variante do BPV-8 (BPV-8-EB) foi identificada em lesões papilomatosas de um bisão europeu na Eslováquia. No Brasil, apesar da infecção pelo BPV ser comumente observada em bovinos, a determinação dos tipos virais associados com a infecção ainda é esporádica. Este estudo tem o objetivo de relatar a ocorrência do BPV-8 no país. A amostra clínica foi obtida em um rebanho de corte do estado do Paraná, região sul do Brasil. O papiloma cutâneo, de aspecto macular e não-verrucoso, estava localizado na região dorsal torácica do animal. A identificação do vírus foi realizada pela reação em cadeia da polimerase (PCR) utilizando primers genéricos para a amplificação parcial do gene L1. O produto amplificado, com aproximadamente 480 pb, foi clonado e os fragmentos presentes em dois clones foram seqüenciados. A comparação da seqüência de nucleotídeos com a de outros papilomavírus demonstrou 100 por cento de identidade com o BPV-8. Assim, esta é a primeira descrição da ocorrência do BPV-8 no Brasil, o que sugere a sua presença nos rebanhos bovinos brasileiros.

Bovine papillomavirus type 4 L1 gene transfection in a Drosophila S2 cell expression system: absence of L1 protein expression

The development of a bovine papillomavirus (BPV) vaccine is an outstanding challenge. BPV protein L1 gene transfection in the Drosophila melanogaster S2 cell expression system failed to produce L1 protein notwithstanding correct L1 gene insertion. Severe genetic inbalance in the host cell line, including cytogenetic alterations, may account for the lack of protein expression.

O desenvolvimento de uma vacina para papilomavirus bovino (BPV) consiste em grande desafio. A transfecção do gene codificante da proteína L1 de BPV em sistema de células S2 de Drosophila melanogaster não logrou sucesso, apesar da correta inserção da seqüência gênica em vetor apropriado.Graves alterações genéticas na linhagem celular S2, que incluem aberrações cromossômicas, provavelmente estão relacionadas à ausência da expressão da proteína desejada.

Viral DNA sequences in peripheral blood and vertical transmission of the virus: a discussion about BPV-1

This study has detected BPV-1 DNA sequences in wart, blood and plasma samples collected from animals affected by papillomatosis, suggesting viral presence inside the cell. We sellected an animal in which we could detect BPV-1 DNA sequences in wart, blood, placenta and amniotic liquid samples and her offspring which presented BPV-1 DNA sequences in blood sample collected immediately after birth. These results show a possible vertical transmission of BPV-1.

Neste estudo detectou-se DNA de BPV-1 em: verruga, sangue e plasma de animais afetados por papilomatose. Estes resultados trazem evidências sobre possível localização intracelular do BPV-1 no sangue. Avaliamos um animal afetado por papilomatose e positivo para BPV-1 em amostras de verruga, sangue, placenta e líquido amniótico e que teve sua cria recém-nascida também positiva para BPV-1 no sangue. Estes resultados indicam possível transmissão vertical do BPV-1.

Bovine papillomavirus type 2 in reproductive tract and gametes of slaughtered bovine females

Papillomaviruses are described selectively infecting epithelial tissues and are associated with many forms of cancer in different species. Considering the widespread dissemination of papillomatosis in livestock, interest is being centred on possible forms of viral transmission and respective mechanisms. In the present study, we report the detection of bovine papillomavirus (BPV) DNA sequences in female reproductive tract tissues, fluids and oocytes from slaughtered bovines not afflicted by cutaneous papillomatosis. BPV-2 DNA sequences were found in ovarian and uterine tissues as well as in oocytes, cumulus cells and uterine flushings. The presence of papillomavirus sequences in reproductive organ tissues and fluids shows that viral infection in organisms can be verified in others tissues, not only in epithelial ones. The present findings alert to the possibility of BPV transmission in embryo transfer programs and assisted fertilization procedures.

Os vírus do papiloma bovino, descritos como agentes infectantes específicos do epitélio, têm sido associados a diversas formas de câncer em diferentes espécies animais. Dada a intensa disseminação da papilomatose nos rebanhos, a investigação de diferentes formas de transmissão e seus respectivos mecanismos tem exigido especial atenção. No presente estudo, é relatada a detecção de seqüências genômicas do papilomavirus bovino (BPV) em ovócitos e tecidos do trato reprodutivo oriundos de fêmeas abatidas comercialmente, não apresentando papilomatose cutânea. A presença de DNA de BPV-2 em tecidos do trato reprodutivo, lavado uterino, ovócitos e células do cumulus traz evidências de que a infecção viral pode se desenvolver fora do tecido epitelial. Esses achados alertam para a possibilidade de transmissão do BPV através dos procedimentos de transferência de embriões e de fertilização in vitro.

Viral DNA sequences in peripheral blood and vertical transmission of the virus: a discussion about BPV-1

This study has detected BPV-1 DNA sequences in wart, blood and plasma samples collected from animals affected by papillomatosis, suggesting viral presence inside the cell. We sellected an animal in which we could detect BPV-1 DNA sequences in wart, blood, placenta and amniotic liquid samples and her offspring which presented BPV-1 DNA sequences in blood sample collected immediately after birth. These results show a possible vertical transmission of BPV-1.

Neste estudo detectou-se DNA de BPV-1 em: verruga, sangue e plasma de animais afetados por papilomatose. Estes resultados trazem evidências sobre possível localização intracelular do BPV-1 no sangue. Avaliamos um animal afetado por papilomatose e positivo para BPV-1 em amostras de verruga, sangue, placenta e líquido amniótico e que teve sua cria recém-nascida também positiva para BPV-1 no sangue. Estes resultados indicam possível transmissão vertical do BPV-1.

Bovine papillomavirus type 2 in reproductive tract and gametes of slaughtered bovine females

Papillomaviruses are described selectively infecting epithelial tissues and are associated with many forms of cancer in different species. Considering the widespread dissemination of papillomatosis in livestock, interest is being centred on possible forms of viral transmission and respective mechanisms. In the present study, we report the detection of bovine papillomavirus (BPV) DNA sequences in female reproductive tract tissues, fluids and oocytes from slaughtered bovines not afflicted by cutaneous papillomatosis. BPV-2 DNA sequences were found in ovarian and uterine tissues as well as in oocytes, cumulus cells and uterine flushings. The presence of papillomavirus sequences in reproductive organ tissues and fluids shows that viral infection in organisms can be verified in others tissues, not only in epithelial ones. The present findings alert to the possibility of BPV transmission in embryo transfer programs and assisted fertilization procedures.

Os vírus do papiloma bovino, descritos como agentes infectantes específicos do epitélio, têm sido associados a diversas formas de câncer em diferentes espécies animais. Dada a intensa disseminação da papilomatose nos rebanhos, a investigação de diferentes formas de transmissão e seus respectivos mecanismos tem exigido especial atenção. No presente estudo, é relatada a detecção de seqüências genômicas do papilomavirus bovino (BPV) em ovócitos e tecidos do trato reprodutivo oriundos de fêmeas abatidas comercialmente, não apresentando papilomatose cutânea. A presença de DNA de BPV-2 em tecidos do trato reprodutivo, lavado uterino, ovócitos e células do cumulus traz evidências de que a infecção viral pode se desenvolver fora do tecido epitelial. Esses achados alertam para a possibilidade de transmissão do BPV através dos procedimentos de transferência de embriões e de fertilização in vitro.

Medico-Surgical Cooperative Treatment of Pulmonary Atresia with Intact Ventricular Septum

PURPOSE: The actual clinical examples of co-appliance of catheter intervention with surgical procedures in the treatment of pulmonary atresia with an intact ventricular septum(PA/IVS) which we have experienced in our institution are here shown, and the anatomical and hemodynamical profiles between each method is compared. METHODS: Medical records of 33 patients with PA/IVS who underwent various treatment from January, 1995 to December, 2000 were reviewed for a retrograde study. RESULTS: In three out of 10 patients who underwent percutaneous balloon pulmonary valvotomy (PPV), residual pulmonary stenosis were observed in their out patient department(OPD) follow-ups, eventually necessitatig balloon pulmonary valvuloplasty(BPV). One out of three patients exhibited deterioration of tricuspid regurgitation after BPV, requiring surgical tricuspid annuloplasty(TAP). Two out of the seven patients who received primarily surgical right ventricle outlet tract(RVOT) repair without any systemic-pulmonary shunt or intervention needed additional intervention employing cardiac catheterization after operation. Two patients received interventional catheterization before surgical RVOT repair. In five out of 11 cases of Fontan type operation, coil embolization of collateral circulation was done before total cavo-pulmonary connection(TCPC), and in three cases, interventional catheterization was needed after TCPC. CONCLUSION: Both medical and surgical treatment modalities are widely used in management of PA/IVS patients, and recent results prove that medico-surgical cooperative treatment is essential.