Canine transmissible venereal tumor - From general to molecular characteristics: A review.

Cancer is a group of complex diseases resulting from the accumulation of genetic and epigenetic changes affecting control and activity of several genes, especially those involved in cell differentiation and growth processes, leading to an abnormal proliferation. When the disease reaches an advanced stage, cancer can lead to metastasis in other organs. Interestingly, recent studies have shown that some types of cancer spread not only through the body, but also can be transmitted among individuals. Therefore, these cancers are known as transmissible tumors. Among the three types of transmissible tumors that occur in nature, the canine transmissible venereal tumor (CTVT) is known as the oldest cancer in the world, since it was originated from a single individual 11 000 years ago. The disease has a worldwide distribution, and its occurrence has been documented since 1810. The CTVT presents three types of cytomorphological classification: lymphocytoid type, mixed type, and plasmacytoid type, the latter being chemoresistant due to overexpression of the ABCB1 gene, and consequently increase of the P-glycoprotein. More knowledge about the epidemiology and evolution of CTVT may help to elucidate the pathway and form of the global spread of the disease.

Epidemiological, clinical, and therapeutic aspects of canine transmissible venereal tumor in Rio de Janeiro, Brazil (2015-2020) / Aspectos epidemiológicos, clínicos e terapêuticos do tumor venéreo transmissível canino no Rio de Janeiro, Brasil (2015-2020)

Canine transmissible venereal tumors (TVT) have a high incidence in Brazil. This is partly due to the large population of stray dogs and the ineffectiveness of epidemiological control programs. This study aimed to describe the epidemiological data, clinical manifestations, and treatments used in dogs affected by TVT. Data were retrospectively collected from the 2015-2020 records of the Veterinary Hospital of the Federal Rural University of Rio de Janeiro. A total of 252 dogs were diagnosed with TVT during the study period. Of these, 81.3% were mixed-breed, 50.4% were males, and 88.9% were young or adult animals. The genital region only was affected in 77.3% of cases. Exclusively extragenital lesions were observed in 22.6% of cases. Among the animals seen, 40.1% received no treatment. Of those treated, 99.3% underwent a vincristine sulfate protocol, and in 77.2%, the treatment resulted in total remission of the neoplasm after 4 to 6 chemotherapy sessions. It was concluded that TVT is a neoplasm most often seen in mixed-breed dogs and located in the genital region, with hemorrhagic secretion being the main clinical sign reported by owners. Vincristine sulfate is currently the most used therapy, with high efficacy. However, despite the good prognosis, there was a high rate of non-adherence or abandonment of treatment, and this is an important factor to be considered and addressed by veterinarians.

O tumor venéreo transmissível (TVT) apresenta elevada incidência no Brasil, relacionada a elevada população de caninos errantes e a ineficácia dos programas de controle epidemiológicos. O objetivo do estudo foi descrever dados epidemiológicos, manifestação clínica e o tratamento empregado em cães acometidos pelo TVT no Hospital Veterinário da Universidade Federal Rural do Rio de Janeiro entre os anos de 2015 e 2020. Foram diagnosticados 252 cães com TVT durante o período do estudo, sendo 81,3% cães sem raça definida, 50,4% machos e 49,6% fêmeas, e com 88,9% animais jovens ou adultos. A região genital foi acometida em 77,3% dos casos. Lesões exclusivamente extragenitais foram observadas em 22,6% dos casos. Quanto ao tratamento, 40,1% dos cães não receberam tratamento. Entre os animais tratados, 99,3% utilizaram protocolo com sulfato de vincristina e em 77,2% o tratamento resultou em remissão total da neoplasia, com a realização de 4 a 6 sessões do quimioterápico. Conclui-se que o TVT é uma neoplasia frequentemente relacionada a cães sem raça definida, localizados na região genital, com secreção hemorrágica sendo o principal sinal clínico reportado pelos proprietários. O sulfato de vincristina é a terapia mais empregada, com alta eficácia. Entretanto, apesar de ser uma neoplasia com bom prognóstico, o alto índice de não adesão ou abandono ao tratamento representa um importante fator a ser considerado e trabalhado pelos médicos veterinários.

Diagnosis of transmissible venereal tumors in bitches - platelet indices are a remarkable marker?

Background: Hematological analyses are seen as more preferred laboratory analyses in canine transmissible venereal tumor studies. There is no information about the availability of platelets and their indices in routine practice in canine transmissible venereal tumor cases. Taking this as a starting point, this study analyzed the usefulness of platelet indices in dogs with transmissible venereal tumor in clinical laboratory diagnosis as well as examined the relationship between white blood cells, red blood cells, platelets (PLT), main platelet volume (MPV), platelet distribution width (PDW), plateletcrit (PCT), and the ratio of main platelet volume to platelets (MPV/PLT). Materials, Methods & Results: In the study, a total of 42 bitches of various breeds were used. Nineteen healthy bitches were used as a control group, and the others 23 with cTVT as a study group. Metastasis was not observed in any of the bitches involved in the study. History, clinical findings, and cytological examinations were evaluated for the diagnosis of cTVT. In animals with hemorrhagic discharge and neoplastic lesions, a vaginal cytological examination was performed. Typical TVT cells with large nuclei and intracytoplasmic vacuoles were observed in the vaginal cytological examinations, and the diagnosis of TVT was made. Healthy bitches (19) and those with TVT (23) were 39.16 5.37 months and 47.61 5.14 months old, respectively. From all animals, 2 mL blood samples were collected from V. cephalica to evaluate PIs in the complete blood count (CBC). Collected blood samples were analyzed using an automated hematology analyzer. As a result of the analysis, WBC, RBC, HGB, HCT, MCV, MCHC, RDW, PLT, MPV, PDW, PCT, and MPV/PLT data were obtained. Mild leukocytosis, an increase in PLT, and a decrease in MCV and MPV/PLT were determined in the study group compared to the control group. Cut-off values in CBC of bitches with TVT were determined as WBC: 13.35 (sensitivity: 78%; specificity: 90%); MCV: 67 (sensitivity: 57%; specificity: 95%); PLT: 315.50 (sensitivity: 65%; specificity: 74%); and MPV/PLT: 0.028 (sensitivity: 78%; specificity: 58%). In CBC analyses, a strong negative correlation between PLT and MPV/PLT was detected in both groups. Discussion: Canine transmissible venereal tumors are common in both stray and pet dogs. It is naturally transferred from animal to animal during mating by live tumor cells. This tumor can commonly affect the external genitalia and internal organs in some cases. It generally has the look of cauliflower, and its surface is ulcerated, inflammatory, hemorrhagic, and infectious. More preferred laboratory analyses are complete blood count and blood chemistry analysis in cTVT for to evaluate the success of treatments. Platelet indices have been investigated in many diseases such as endotoxemia, chronic enteropathy, mammary tumor, parvoviral enteritis, septic peritonitis, lymphoma, pyometra, visceral leishmaniasis, and babesiosis in dogs. There is no information available for either diagnostic or prognostic use of the PIs in canine TVT cases. Ultimately, in light of the presented study's results, platelet indices, especially PLT and the MPV/PLT ratio, seem to be notable laboratory markers in terms of easy accessibility and low-cost assessment techniques in canine transmissible venereal tumor cases. New data, however, should be established by a thorough follow-up study using a larger sample size and addressing its usefulness as a diagnostic or prognostic marker in canine transmissible venereal tumors.

In vitro chemosensitivity of a canine tumor venereal transmissible cancer cell line.

The canine transmissible venereal tumor (CTVT) is the most common malignity in dogs. Because there are reports that this tumor is resistant to vincristine sulfate, the chemotherapeutic options are scarce, and the development of new therapeutic approaches is necessary. In this study, we evaluated the cytotoxic activity of vincristine, doxorubicin, temozolomide, panobinostat, toceranib, gemcitabine, cisplatin, fluorouracil, cyclophosphamide, and methotrexate on a CTVT cell line, determining that all drugs decreased the viability in a dose-dependent manner. Furthermore, they inhibit cellular migration in a time- and drug-dependent manner, as evaluated by the wound healing assay. On the other hand, vincristine, panobinostat, gemcitabine, toceranib, cyclophosphamide, and methotrexate increased the percentage of cells in the subG1 phase, and doxorubicin, temozolomide, gemcitabine, toceranib, and methotrexate decreased the percentage of cells in the synthesis phase. To efficientize the use of vincristine, only toceranib increased the cytotoxic effect of vincristine in a synergistic manner. Our results confirm the use of vincristine as the gold standard for CTVT treatment as monotherapy and suggest the use of a combinatorial and sequential treatment with toceranib.

Could dysregulation of RASSF1 expression be a mechanism of tumorigenesis in CTVT? / A desregulação da expressão de RASSF1 pode ser um mecanismo da tumorigênese em TVTC?

Canine transmissible venereal tumor (CTVT) is the oldest known somatic cell lineage. It is a transmissible cancer that propagates naturally in dogs and reportedly contains gene mutations. RASSF1 participates in DNA damage repair, and its downregulation, results in tumor progression. Hence, RASSF1 is a tumor suppressor gene. Its expression was quantified in tumors from seventeen animals and three cell cultures derived from tumors. In general, RASSF1 was underexpressed in 65%, and absent in 35% of tumor samples. Cells from tumor tissue cultures showed decreased expression of RASSF1 in 67% and elevated expression in 33% of samples tested. The tumor tissues showed significantly lower levels of RASSF1 expression compared to cultured cells. Previously we reported that both the tumor microenvironment and the host immune system appear to influence the tumorigenesis and stage of CTVT. This is the first article to demonstrate the expression of RASSF1 in CTVT. Decreased RASSF1 possibly helps tumor progression.

O tumor venéreo transmissível canino (TVTC) é a linhagem de células somáticas mais antiga conhecida. É um câncer transmissível que se propaga naturalmente em cães e mutações genéticas já foram relatadas. O gene RASSF1 atua no reparo de danos ao DNA e presume-se que, quando suprimido ou com expressão gênica reduzida, o TVTC tende a progredir. A expressão do gene supressor de tumor, como RASSF1, foi quantificada em tecidos de dezessete animais e três culturas de células de tecidos tumorais. Em geral, o gene RASSF1 apresentou prevalência de subexpressão (65%) e ausência em 35% dos demais tecidos analisados. Células isoladas de culturas de tecidos tumorais também demonstraram 67% com expressão diminuída e 33% com expressão elevada, com diferença significativa entre os níveis de expressão gênica em amostras de tecido quando comparadas às culturas de células, com tecidos apresentando níveis mais baixos de expressão gênica em comparação com células. Anteriormente, relatamos que tanto o microambiente tumoral quanto o sistema imunológico do hospedeiro parecem influenciar a tumorigênese e o estágio do TVTC. Este é o primeiro artigo a demonstrar a expressão de RASSF1 no TVTC, possivelmente alterando sua tumorigênese e auxiliando no aumento da progressão tumoral.

Epidemiological study of canine transmissible venereal tumor (CTVT) in Brazil, 2000-2020.

Canine transmissible venereal tumor (CTVT) is a contagious neoplasm, mainly transmitted through coitus. This round cell mesenchymal tumor is common in Brazil, often located in the genitalia although extragenital presentations may also occur, such as cutaneous, oral, and nasal forms. The objective of this study was to perform an epidemiological analysis of CTVT from published data in the recent academic literature to systematically demonstrate the distribution of CTVT in Brazil, identify the frequency of this neoplasm and its main diagnostic tests, and characterize its main clinical manifestations in Brazil. For such purpose, it was analyzed the scientific publications with cases of CTVT in Brazil, in English or Portuguese, published between 2000-2020. The CTVT was identified in 19 Brazilian states plus the Federal District, totaling 3,622 cases across the national territory, with the largest number of cases recorded in the Southeast region. The cytological exam was the most used for the diagnosis of CTVT (89.2 %), followed by histopathological (37.8 %) and immunohistochemistry (13.5 %)1 . Predominant epidemiological aspects of CTVT identified in the study were: Mixed breed dogs (75.2 %), females (62.5 %), in adulthood (between 2 and 7 years) and dogs with free extra outdoor access (91.1 %). Genital presentation was the most frequent in the literature (86 %), followed by cutaneous (21.8 %), nasal (10 %), oral and lymph nodes presentations (10-5 %) and less frequent manifestations as ocular and anal/perianal (< 5 %). CTVT is a neoplasm widely distributed in Brazil, highly frequent and with several forms of clinical presentation, which can be underdiagnosed if there is no adequate knowledge of this tumor and its epidemiological characteristics. The extragenital manifestations of the neoplasm need further studies for its better characterization and more precise definition of its frequencies.

Comparison between hematological and biochemical changes caused by conventional and metronomic chemotherapies in the treatment of canine transmissible venereal tumor

ABSTRACT: This study aimed to report the hematological and biochemical changes caused by conventional and metronomic chemotherapies, using vincristine sulfate to treat canine Transmissible Venereal Tumor (TVT). Twelve dogs were selected, six of them for the group receiving conventional chemotherapy (G1), and six dogs for the group receiving metronomic chemotherapy (G2). The G1 group received vincristine sulfate once a week at the dose of 0.75mg/m² until the tumor had disappeared with treatment, and the G2 group was treated with vincristine sulfate three times a week at the dose of 0.25mg/m2 until the tumor had disappeared. Before and after chemotherapy treatment, hematological and biochemical blood tests were performed to evaluate the main alterations caused by both chemotherapeutic models. Dogs undergoing conventional chemotherapy had significant leukocyte changes (p 0.05), causing neutropenia and leukopenia. In dogs undergoing metronomic chemotherapy, leukocytes remained within the reference range. Half of the dogs in group G1 had normochromic, normocytic anemia. The only biochemical alteration observed was the increase of urea in group G2. Thus, metronomic chemotherapy for the treatment of TVT with vincristine sulfate proved to be an excellent method for treatment, with fewer adverse effects, especially in maintaining the leukogram of dogs within normal range and reducing the number of anemia in animals during treatment.

RESUMO: Esta pesquisa teve como objetivo relatar as alterações hematológicas e bioquímicas causadas pelo tratamento quimioterápico convencional e pela quimioterapia metronômica, utilizando-se sulfato de vincristina para o tratamento do tumor venéreo transmissível canino(TVTC). Foram selecionados 12 cães, sendo seis para o grupo que recebeu quimioterapia convencional (G1) e seis cães para o grupo que recebeu quimioterapia metronômica (G2). O grupo G1 recebeu sulfato de vincristina, uma vez por semana, na dose de 0,75mg/m2, até o desaparecimento do tumor e o grupo G2 foi tratado com sulfato de vincristina, três vezes por semana, na dose de 0,25mg/m2, até o desaparecimento do tumor. Antes e após o tratamento quimioterápico foram realizados exames hematológicos e bioquímicos sanguíneos para avaliação das principais alterações causadas pelos dois modelos quimioterápicos. Os cães submetidos à quimioterapia convencional tiveram alterações leucocitárias significativas (p 0,05), causando uma leucopenia por neutropenia enquanto nos cães, submetidos à quimioterapia metronômica, os leucócitos mantiveram-se dentro do intervalo de referência. A metade dos cães do grupo G1 tiveram uma anemia do tipo normocítica normocrômica. A única alteração bioquímica observada foi o aumento da ureia no grupo G2. Desta forma, a quimioterapia metronômica para o tratamento do TVT com sulfato de vincristina, demonstrou ser um excelente método para a cura do animal, com menores reduções de efeitos adversos, sobretudo, na manutenção do leucograma dos cães e na redução de animais com anemia.

Comparison between hematological and biochemical changes caused by conventional and metronomic chemotherapies in the treatment of canine transmissible venereal tumor / Comparação entre as alterações hematológicas e bioquímicas ocasionadas pela quimioterapia convencional e quimioterapia metronômica no tratamento do tumor venéreo transmissível canino

This study aimed to report the hematological and biochemical changes caused by conventional and metronomic chemotherapies, using vincristine sulfate to treat canine Transmissible Venereal Tumor (TVT). Twelve dogs were selected, six of them for the group receiving conventional chemotherapy (G1), and six dogs for the group receiving metronomic chemotherapy (G2). The G1 group received vincristine sulfate once a week at the dose of 0.75mg/m² until the tumor had disappeared with treatment, and the G2 group was treated with vincristine sulfate three times a week at the dose of 0.25mg/m2 until the tumor had disappeared. Before and after chemotherapy treatment, hematological and biochemical blood tests were performed to evaluate the main alterations caused by both chemotherapeutic models. Dogs undergoing conventional chemotherapy had significant leukocyte changes (p<0.05), causing neutropenia and leukopenia. In dogs undergoing metronomic chemotherapy, leukocytes remained within the reference range. Half of the dogs in group G1 had normochromic, normocytic anemia. The only biochemical alteration observed was the increase of urea in group G2. Thus, metronomic chemotherapy for the treatment of TVT with vincristine sulfate proved to be an excellent method for treatment, with fewer adverse effects, especially in maintaining the leukogram of dogs within normal range and reducing the number of anemia in animals during treatment.(AU)

Esta pesquisa teve como objetivo relatar as alterações hematológicas e bioquímicas causadas pelo tratamento quimioterápico convencional e pela quimioterapia metronômica, utilizando-se sulfato de vincristina para o tratamento do tumor venéreo transmissível canino(TVTC). Foram selecionados 12 cães, sendo seis para o grupo que recebeu quimioterapia convencional (G1) e seis cães para o grupo que recebeu quimioterapia metronômica (G2). O grupo G1 recebeu sulfato de vincristina, uma vez por semana, na dose de 0,75mg/m2, até o desaparecimento do tumor e o grupo G2 foi tratado com sulfato de vincristina, três vezes por semana, na dose de 0,25mg/m2, até o desaparecimento do tumor. Antes e após o tratamento quimioterápico foram realizados exames hematológicos e bioquímicos sanguíneos para avaliação das principais alterações causadas pelos dois modelos quimioterápicos. Os cães submetidos à quimioterapia convencional tiveram alterações leucocitárias significativas (p<0,05), causando uma leucopenia por neutropenia enquanto nos cães, submetidos à quimioterapia metronômica, os leucócitos mantiveram-se dentro do intervalo de referência. A metade dos cães do grupo G1 tiveram uma anemia do tipo normocítica normocrômica. A única alteração bioquímica observada foi o aumento da ureia no grupo G2. Desta forma, a quimioterapia metronômica para o tratamento do TVT com sulfato de vincristina, demonstrou ser um excelente método para a cura do animal, com menores reduções de efeitos adversos, sobretudo, na manutenção do leucograma dos cães e na redução de animais com anemia.(AU)

Comparison between hematological and biochemical changes caused by conventional and metronomic chemotherapies in the treatment of canine transmissible venereal tumor / Comparação entre as alterações hematológicas e bioquímicas ocasionadas pela quimioterapia convencional e quimioterapia metronômica no tratamento do tumor venéreo transmissível canino

This study aimed to report the hematological and biochemical changes caused by conventional and metronomic chemotherapies, using vincristine sulfate to treat canine Transmissible Venereal Tumor (TVT). Twelve dogs were selected, six of them for the group receiving conventional chemotherapy (G1), and six dogs for the group receiving metronomic chemotherapy (G2). The G1 group received vincristine sulfate once a week at the dose of 0.75mg/m² until the tumor had disappeared with treatment, and the G2 group was treated with vincristine sulfate three times a week at the dose of 0.25mg/m2 until the tumor had disappeared. Before and after chemotherapy treatment, hematological and biochemical blood tests were performed to evaluate the main alterations caused by both chemotherapeutic models. Dogs undergoing conventional chemotherapy had significant leukocyte changes (p<0.05), causing neutropenia and leukopenia. In dogs undergoing metronomic chemotherapy, leukocytes remained within the reference range. Half of the dogs in group G1 had normochromic, normocytic anemia. The only biochemical alteration observed was the increase of urea in group G2. Thus, metronomic chemotherapy for the treatment of TVT with vincristine sulfate proved to be an excellent method for treatment, with fewer adverse effects, especially in maintaining the leukogram of dogs within normal range and reducing the number of anemia in animals during treatment.(AU)

Esta pesquisa teve como objetivo relatar as alterações hematológicas e bioquímicas causadas pelo tratamento quimioterápico convencional e pela quimioterapia metronômica, utilizando-se sulfato de vincristina para o tratamento do tumor venéreo transmissível canino(TVTC). Foram selecionados 12 cães, sendo seis para o grupo que recebeu quimioterapia convencional (G1) e seis cães para o grupo que recebeu quimioterapia metronômica (G2). O grupo G1 recebeu sulfato de vincristina, uma vez por semana, na dose de 0,75mg/m2, até o desaparecimento do tumor e o grupo G2 foi tratado com sulfato de vincristina, três vezes por semana, na dose de 0,25mg/m2, até o desaparecimento do tumor. Antes e após o tratamento quimioterápico foram realizados exames hematológicos e bioquímicos sanguíneos para avaliação das principais alterações causadas pelos dois modelos quimioterápicos. Os cães submetidos à quimioterapia convencional tiveram alterações leucocitárias significativas (p<0,05), causando uma leucopenia por neutropenia enquanto nos cães, submetidos à quimioterapia metronômica, os leucócitos mantiveram-se dentro do intervalo de referência. A metade dos cães do grupo G1 tiveram uma anemia do tipo normocítica normocrômica. A única alteração bioquímica observada foi o aumento da ureia no grupo G2. Desta forma, a quimioterapia metronômica para o tratamento do TVT com sulfato de vincristina, demonstrou ser um excelente método para a cura do animal, com menores reduções de efeitos adversos, sobretudo, na manutenção do leucograma dos cães e na redução de animais com anemia.(AU)

Ocorrência de tumor venéreo transmissível em cães atendidos no Hovet - PA UFRA / Occurrence of transmissible venereum tumor in dogs served at hove t- PA UFRA

The objective was to verify the occurrence of the transmissible venereal tumor in canines attended at (UFRA), Campus Belém / Pará, from March 2016 to September 2019. The information was obtained from the SISVET® program (Veterinary System). A total of 3,714 records were analyzed regarding the animals served and sent to the animal reproduction sector. Of this total, 155 were from dogs diagnosed with transmissible venereal tumor and 8 negative cases, 102 females and 53 males. The data were tabulated in an electronic spreadsheet, submitted to statistical analysis using the SAS® Universty Edition software regarding the frequency of occurrences, the influence of the age group, sex, race, through the Chi-Square test with 5% significance. Of the 155 cases of TVT diagnosed, 102 (65.81%) were in females and 53 (34.19%) males, with prevalence in SRD (72.86%) and in adults (54.19%). As for the site of involvement, 32.26% were in vulva and 20% in the body of the penis. Regarding the neighborhoods, there was a predominance in Guamá with 25 cases (15.59%). Thus, it is concluded that TVT was more frequent in SRD females and in adult dogs. Vulva 50 (32.26%), vagina 46 (29.68%) and penis 31 (20%) were the most affected sites.

Ocorrência de tumor venéreo transmissível em cães atendidos no Hovet - PA UFRA / Occurrence of transmissible venereum tumor in dogs served at hove t- PA UFRA

The objective was to verify the occurrence of the transmissible venereal tumor in canines attended at (UFRA), Campus Belém / Pará, from March 2016 to September 2019. The information was obtained from the SISVET® program (Veterinary System). A total of 3,714 records were analyzed regarding the animals served and sent to the animal reproduction sector. Of this total, 155 were from dogs diagnosed with transmissible venereal tumor and 8 negative cases, 102 females and 53 males. The data were tabulated in an electronic spreadsheet, submitted to statistical analysis using the SAS® Universty Edition software regarding the frequency of occurrences, the influence of the age group, sex, race, through the Chi-Square test with 5% significance. Of the 155 cases of TVT diagnosed, 102 (65.81%) were in females and 53 (34.19%) males, with prevalence in SRD (72.86%) and in adults (54.19%). As for the site of involvement, 32.26% were in vulva and 20% in the body of the penis. Regarding the neighborhoods, there was a predominance in Guamá with 25 cases (15.59%). Thus, it is concluded that TVT was more frequent in SRD females and in adult dogs. Vulva 50 (32.26%), vagina 46 (29.68%) and penis 31 (20%) were the most affected sites.(AU)

Presence of Papillomavirus DNA sequences in the canine transmissible venereal tumor (CTVT).

BACKGROUND: The canine transmissible venereal tumor (CTVT) or Sticker's sarcoma is a neoplastic disease affecting dogs. This disease is presented as a tumoral mass in the genital organs of both, male and female individuals. Up to date, there is no clear evidence indicating a viral agent as the causative mediator for CTVT development. PURPOSE: The present work aims to analyze 21 samples from canines with CTVT for molecular identification of Papillomavirus DNA sequences. In addition, microbiological analysis, cytologic and histopathologic evaluations were also performed. RESULTS: All patients showed no biochemical and microbiological alterations. Molecular analysis demonstrated the viral DNA presence in the samples using different primer sets. The MY primers amplified a 450 bp band in seven out of 21 samples (33%). The PVF and Fap64 primer set, targeting the L1 sequence of Canine Papillomavirus (CPV), showed positivity in 16 out of 21 samples (76%). CONCLUSION: These results support the possible causative association between CPV and CTVT; nevertheless, additional studies are required to uphold such statement. This work presents evidence indicating that a viral agent might be involved in the pathogenesis of CTVT and set the bases for a better understanding of the CTVT pathobiology.

Multiple metastases of a transmissible venereal tumor in a dog

Background: Transmissible venereal tumor (TVT) is a common contagious neoplasm in dogs that spreads through coitus.Extra-genital presentations of this tumor are frequent and usually develop through implantation of neoplastic cells onexposed mucosae. TVT metastasis is rare, and when it happens its usually affecting regional lymph nodes and adjacentcutaneous tissue.Case: A female mixed breed dog, with estimated age of 7 to 11 months old, was rescued from the streets and taken to aveterinary clinic in the city of Porto Alegre, RS. The animal had multiple nodules on its body, vulva, ocular mucosa, andgingiva, along with signs of malnutrition and apathy. Cytological examination of the nodules and vulva was done andyielded a cytologic picture compatible with TVT. Weakly treatment with 0,3mg/m² vincristine sulphate was used untilclinical cure was noted. Approximately two weeks after clinical cure, the dog showed a blue colored eye and was referredfor ophthalmological, where it was diagnosed with vision loss due to glaucoma secondary to a neoplasm. The eye wasthen removed and sent for histopathological evaluation. Histopathology of the eye was compatible with TVT diagnosis.One month after enucleation the animal display dispenia, pain, aggressiveness and epistaxis. The animal was euthanizedand submitted for post-mortem evaluation. At necropsy there was a well-defined grayish-white, nodule near the thalamus.Similar nodules were also found on the lung, and anterior chamber of the eye. Histologically, all the nodules were compatible with TVT. Immunohistochemical examination was done, with the neoplastic cells being positive for vimentin andnegative for cytokeratin, CD79a, CD3 and CD117. Based on the post-mortem examination and clinical history, diagnosisof TVT...(AU)

Multiple metastases of a transmissible venereal tumor in a dog

Background: Transmissible venereal tumor (TVT) is a common contagious neoplasm in dogs that spreads through coitus.Extra-genital presentations of this tumor are frequent and usually develop through implantation of neoplastic cells onexposed mucosae. TVT metastasis is rare, and when it happens it’s usually affecting regional lymph nodes and adjacentcutaneous tissue.Case: A female mixed breed dog, with estimated age of 7 to 11 months old, was rescued from the streets and taken to aveterinary clinic in the city of Porto Alegre, RS. The animal had multiple nodules on its body, vulva, ocular mucosa, andgingiva, along with signs of malnutrition and apathy. Cytological examination of the nodules and vulva was done andyielded a cytologic picture compatible with TVT. Weakly treatment with 0,3mg/m² vincristine sulphate was used untilclinical cure was noted. Approximately two weeks after clinical cure, the dog showed a blue colored eye and was referredfor ophthalmological, where it was diagnosed with vision loss due to glaucoma secondary to a neoplasm. The eye wasthen removed and sent for histopathological evaluation. Histopathology of the eye was compatible with TVT diagnosis.One month after enucleation the animal display dispenia, pain, aggressiveness and epistaxis. The animal was euthanizedand submitted for post-mortem evaluation. At necropsy there was a well-defined grayish-white, nodule near the thalamus.Similar nodules were also found on the lung, and anterior chamber of the eye. Histologically, all the nodules were compatible with TVT. Immunohistochemical examination was done, with the neoplastic cells being positive for vimentin andnegative for cytokeratin, CD79a, CD3 and CD117. Based on the post-mortem examination and clinical history, diagnosisof TVT...

Immunogenic potential of three transmissible venereal tumor cell lysates to prime canine-dendritic cells for cancer immunotherapy.

Whole tumor cell lysates consist of a mixture of tumor antigens and danger associated molecular patterns (DAMPs) that can be used for dendritic cell maturation and consequently for the activation of a polyclonal T cell-specific tumor response. We evaluated the in vitro efficacy of three different preparations of canine transmissible venereal tumor (TVT) cell lysates: hypochlorous acid-whole tumor cell lysates (HOCl-L), heat shock-whole tumor cell lysates (HS-L), and freeze-thaw cycles-whole tumor cell lysates (FT-L) for the maturation of canine-derived dendritic cells. Our results showed calreticulin, HSP70, and HSP90 release in the three tumor lysates preparations (HOCl-L, HS-L, and FT-L); however, HMGB1 was detected only in HOCl-L and FT-L. Additionally, the uptake by HOCl-L pulsed dendritic cell (DC) increased compared to HS-L and FT-L pulsed DC; and dendritic cell maturation was confirmed by the appropriate cell surface markers (CD11c, CD80, CD83, and MHCII). Furthermore, dendritic cells pulsed with HOCl-L, HS-L or FT-L were cultured with canine lymphocytes. There was an increase of Th1-type cytokines (IL-12, TNF-α, and IFN-γ), in all the tumor cell lysates co-cultures, this correlates with T lymphocyte activation and cytotoxic response. Our data confirm that TVT cell lysates can induce functional canine-DC and that HOCl-L is the most effective one. This preparation of TVT cell lysates with HOCl is an attractive approach that allows the recognition of neoantigens as potential tumor targets and DC priming and therefore could be used for cancer immunotherapy against TVT.

Immunohistochemical, lectin histochemical and ultrastructural studies of canine transmissible venereal tumor in Brazil / Estudos imuno-histoquímico, lectino-histoquímico e ultraestrutural do Tumor Venéreo Transmissível Canino no Brasil

Canine transmissible venereal tumor (CTVT) is a naturally occurring contagious round-cell neoplasia, with poorly understood origin and transmission. This study aims to further investigate the tumor nature through immunohistochemistry, lectin histochemistry and transmission electron microscopy (TEM) analysis, and to provide support for diagnostic and differential diagnoses of CTVT. Immunohistochemistry was performed in 10 genital and six exclusively extragenital tumors, which were previously diagnosed by citology and histopathology. CTVT samples were incubated with biotinylated antibodies to specific membrane and cytoplasmic antigens (anti-lysozyme, anti-macrophage, anti-vimentin, anti-CD18, monoclonal anti-CD117, monoclonal anti-CD3, polyclonal anti-CD117, polyclonal CD3 and anti-CD79a), followed by the avidin-biotin-peroxidase complex technique. The lectins Con A, DBA, SBA, PNA, UEA-1, WGA, sWGA, GSL, JSA, PSA, PHA-L, PHA-E and RCA were additionally tested in four genital CTVTs and TEM was performed in eight genital tumors. The anti-vimentin antibody revealed strong immunoreactivity to neoplastic cells in all the assessed samples (16/16). The polyclonal anti-CD3 antibodies showed moderate to strong immunoreactivity in fourteen (14/16) and the polyclonal anti-CD117 in fifteen cases (15/16). There was no immunoreactivity to anti-lysozyme, anti-macrophage, anti-CD18, monoclonal anti-CD117, monoclonal anti-CD3 and anti-CD79a antibodies. At lectin histochemistry, it was observed strong staining of tumor cells to Con-A, PHA-L and RCA. There was no histopathological and immunoreactivity differences between genital and extragenital CTVTs. These findings do not support the hypothesis of histiocytic origin of CTVT. In contrast, the lectin histochemical results were similar to cells from lymphoid/myeloid origin.(AU)

O Tumor Venéreo Transmissível Canino (CTVT) é uma neoplasia de células células redondas, contagiosa, com origem e transmissão ainda mal compreendidas. Com a finalidade de aprofundar a investigação sobre a natureza (origem) do TVTC, bem como fornecer subsídios para o estabelecimento do diagnóstico e diagnóstico diferencial, realizaram-se avaliações imuno-histoquímica, lectino-histoquímica e ultraestrutural de TVTC(s). A avaliação imuno-histoquímica foi feita em 10 TVTCs genitais e em 6 exclusivamente extragenitais previamente diagnosticados através de citologia e da histopatologia. Os TVTCs foram testados para reagentes específicos de antígenos de membrana e citoplasmáticos (anti-lisozima, anti-macrófago, anti-vimentina, anti-CD18, anti-CD3, anti-CD79, anti-CD117) com utilização da técnica complexo avidina-biotina-peroxidase. Adicionalmente, foram utilizadas as lectinas Con A, DBA, SBA, PNA, UEA-1, WGA, sWGA, GSL, SJA, PSA, PHA-L, PHA-E e RCA em quatro TVTCs genitais. Microscopia eletrônica foi realizada em oito TVTC genitais. Em 100% dos tumores testados (16/16) com anticorpo anti-vimentina (mono e policlonal) houve forte imuno-reatividade. Não houve reatividade para os anticorpos anti-lisozima, anti-macrófago, anti-CD18, anti-CD3, anti-CD79a e anti-CD117 quando empregamos anticorpos monoclonais, entretanto, com a utilização de anticorpos policlonais verificou-se marcação dos tumores com os anticorpos anti-CD3 e anti-CD117. Na avaliação lectino-histoquímica foi verificada forte marcação das células tumorais com Con-A, PHA-L e RCA. Não houve diferença histopatológica e de imuno-reatividade entre os TVTCs genitais e extragenitais. Estes achados não corroboram com a hipótese da origem histiocítica do CTVT (ausência de reatividade dos anticorpos anti-lisozima, anti-macrófago e anti-CD18), entretanto, os resultados da avaliação lectino-histoquímica foram em parte similares aos obtidos quando células de origem linfóide/mielóide (ConA, PHA-L e RCA) foram analisadas (Gimeno et al. 1995).(AU)

Immunohistochemical, lectin histochemical and ultrastructural studies of canine transmissible venereal tumor in Brazil / Estudos imuno-histoquímico, lectino-histoquímico e ultraestrutural do Tumor Venéreo Transmissível Canino no Brasil

Canine transmissible venereal tumor (CTVT) is a naturally occurring contagious round-cell neoplasia, with poorly understood origin and transmission. This study aims to further investigate the tumor nature through immunohistochemistry, lectin histochemistry and transmission electron microscopy (TEM) analysis, and to provide support for diagnostic and differential diagnoses of CTVT. Immunohistochemistry was performed in 10 genital and six exclusively extragenital tumors, which were previously diagnosed by citology and histopathology. CTVT samples were incubated with biotinylated antibodies to specific membrane and cytoplasmic antigens (anti-lysozyme, anti-macrophage, anti-vimentin, anti-CD18, monoclonal anti-CD117, monoclonal anti-CD3, polyclonal anti-CD117, polyclonal CD3 and anti-CD79a), followed by the avidin-biotin-peroxidase complex technique. The lectins Con A, DBA, SBA, PNA, UEA-1, WGA, sWGA, GSL, JSA, PSA, PHA-L, PHA-E and RCA were additionally tested in four genital CTVTs and TEM was performed in eight genital tumors. The anti-vimentin antibody revealed strong immunoreactivity to neoplastic cells in all the assessed samples (16/16). The polyclonal anti-CD3 antibodies showed moderate to strong immunoreactivity in fourteen (14/16) and the polyclonal anti-CD117 in fifteen cases (15/16). There was no immunoreactivity to anti-lysozyme, anti-macrophage, anti-CD18, monoclonal anti-CD117, monoclonal anti-CD3 and anti-CD79a antibodies. At lectin histochemistry, it was observed strong staining of tumor cells to Con-A, PHA-L and RCA. There was no histopathological and immunoreactivity differences between genital and extragenital CTVTs. These findings do not support the hypothesis of histiocytic origin of CTVT. In contrast, the lectin histochemical results were similar to cells from lymphoid/myeloid origin.(AU)

O Tumor Venéreo Transmissível Canino (CTVT) é uma neoplasia de células células redondas, contagiosa, com origem e transmissão ainda mal compreendidas. Com a finalidade de aprofundar a investigação sobre a natureza (origem) do TVTC, bem como fornecer subsídios para o estabelecimento do diagnóstico e diagnóstico diferencial, realizaram-se avaliações imuno-histoquímica, lectino-histoquímica e ultraestrutural de TVTC(s). A avaliação imuno-histoquímica foi feita em 10 TVTCs genitais e em 6 exclusivamente extragenitais previamente diagnosticados através de citologia e da histopatologia. Os TVTCs foram testados para reagentes específicos de antígenos de membrana e citoplasmáticos (anti-lisozima, anti-macrófago, anti-vimentina, anti-CD18, anti-CD3, anti-CD79, anti-CD117) com utilização da técnica complexo avidina-biotina-peroxidase. Adicionalmente, foram utilizadas as lectinas Con A, DBA, SBA, PNA, UEA-1, WGA, sWGA, GSL, SJA, PSA, PHA-L, PHA-E e RCA em quatro TVTCs genitais. Microscopia eletrônica foi realizada em oito TVTC genitais. Em 100% dos tumores testados (16/16) com anticorpo anti-vimentina (mono e policlonal) houve forte imuno-reatividade. Não houve reatividade para os anticorpos anti-lisozima, anti-macrófago, anti-CD18, anti-CD3, anti-CD79a e anti-CD117 quando empregamos anticorpos monoclonais, entretanto, com a utilização de anticorpos policlonais verificou-se marcação dos tumores com os anticorpos anti-CD3 e anti-CD117. Na avaliação lectino-histoquímica foi verificada forte marcação das células tumorais com Con-A, PHA-L e RCA. Não houve diferença histopatológica e de imuno-reatividade entre os TVTCs genitais e extragenitais. Estes achados não corroboram com a hipótese da origem histiocítica do CTVT (ausência de reatividade dos anticorpos anti-lisozima, anti-macrófago e anti-CD18), entretanto, os resultados da avaliação lectino-histoquímica foram em parte similares aos obtidos quando células de origem linfóide/mielóide (ConA, PHA-L e RCA) foram analisadas (Gimeno et al. 1995).(AU)

Immunohistochemical, lectin histochemical and ultrastructural studies of canine transmissible venereal tumor in Brazil

ABSTRACT: Canine transmissible venereal tumor (CTVT) is a naturally occurring contagious round-cell neoplasia, with poorly understood origin and transmission. This study aims to further investigate the tumor nature through immunohistochemistry, lectin histochemistry and transmission electron microscopy (TEM) analysis, and to provide support for diagnostic and differential diagnoses of CTVT. Immunohistochemistry was performed in 10 genital and six exclusively extragenital tumors, which were previously diagnosed by citology and histopathology. CTVT samples were incubated with biotinylated antibodies to specific membrane and cytoplasmic antigens (anti-lysozyme, anti-macrophage, anti-vimentin, anti-CD18, monoclonal anti-CD117, monoclonal anti-CD3, polyclonal anti-CD117, polyclonal CD3 and anti-CD79a), followed by the avidin-biotin-peroxidase complex technique. The lectins Con A, DBA, SBA, PNA, UEA-1, WGA, sWGA, GSL, JSA, PSA, PHA-L, PHA-E and RCA were additionally tested in four genital CTVTs and TEM was performed in eight genital tumors. The anti-vimentin antibody revealed strong immunoreactivity to neoplastic cells in all the assessed samples (16/16). The polyclonal anti-CD3 antibodies showed moderate to strong immunoreactivity in fourteen (14/16) and the polyclonal anti-CD117 in fifteen cases (15/16). There was no immunoreactivity to anti-lysozyme, anti-macrophage, anti-CD18, monoclonal anti-CD117, monoclonal anti-CD3 and anti-CD79a antibodies. At lectin histochemistry, it was observed strong staining of tumor cells to Con-A, PHA-L and RCA. There was no histopathological and immunoreactivity differences between genital and extragenital CTVTs. These findings do not support the hypothesis of histiocytic origin of CTVT. In contrast, the lectin histochemical results were similar to cells from lymphoid/myeloid origin.

RESUMO: O Tumor Venéreo Transmissível Canino (CTVT) é uma neoplasia de células células redondas, contagiosa, com origem e transmissão ainda mal compreendidas. Com a finalidade de aprofundar a investigação sobre a natureza (origem) do TVTC, bem como fornecer subsídios para o estabelecimento do diagnóstico e diagnóstico diferencial, realizaram-se avaliações imuno-histoquímica, lectino-histoquímica e ultraestrutural de TVTC(s). A avaliação imuno-histoquímica foi feita em 10 TVTCs genitais e em 6 exclusivamente extragenitais previamente diagnosticados através de citologia e da histopatologia. Os TVTCs foram testados para reagentes específicos de antígenos de membrana e citoplasmáticos (anti-lisozima, anti-macrófago, anti-vimentina, anti-CD18, anti-CD3, anti-CD79, anti-CD117) com utilização da técnica complexo avidina-biotina-peroxidase. Adicionalmente, foram utilizadas as lectinas Con A, DBA, SBA, PNA, UEA-1, WGA, sWGA, GSL, SJA, PSA, PHA-L, PHA-E e RCA em quatro TVTCs genitais. Microscopia eletrônica foi realizada em oito TVTC genitais. Em 100% dos tumores testados (16/16) com anticorpo anti-vimentina (mono e policlonal) houve forte imuno-reatividade. Não houve reatividade para os anticorpos anti-lisozima, anti-macrófago, anti-CD18, anti-CD3, anti-CD79a e anti-CD117 quando empregamos anticorpos monoclonais, entretanto, com a utilização de anticorpos policlonais verificou-se marcação dos tumores com os anticorpos anti-CD3 e anti-CD117. Na avaliação lectino-histoquímica foi verificada forte marcação das células tumorais com Con-A, PHA-L e RCA. Não houve diferença histopatológica e de imuno-reatividade entre os TVTCs genitais e extragenitais. Estes achados não corroboram com a hipótese da origem histiocítica do CTVT (ausência de reatividade dos anticorpos anti-lisozima, anti-macrófago e anti-CD18), entretanto, os resultados da avaliação lectino-histoquímica foram em parte similares aos obtidos quando células de origem linfóide/mielóide (ConA, PHA-L e RCA) foram analisadas (Gimeno et al. 1995).

Clinicopathological evaluation of disseminated metastases of transmissible venereal tumor in a spayed bitch

Background: Although transmissible venereal tumor (TVT, transmissible venereal sarcoma, Stickers sarcoma) that affects dogs and other canids can be seen in many countries, it especially emerges in the countries which homeless dog population is very high. Female dogs are more susceptible than males. Transmissible venereal tumor is usually transmitted to genital organs during coitus and occasionally by social behavior such as sniffing and licking. The tumor is generally observed in the posterior part of the vagina. The tumor usually appears in various sizes, in the appearance of cauliflower, red and fragile. Metastases are rarely reported in cases with TVT. Metastases have been detected in lung, liver, tonsils, skin, lymph nodes, muscles, spleen. The diagnosis of transmissible venereal tumor is achived by considering the history of the animal, gross lesions, cytological examination and histopathology. Chemotherapy is frequently used in the treatment of TVT. In addition, radiotherapy, cryosurgery, surgical incision and immunotherapy are rarely applied for treatment. Chemical agents such as doxorubicin, vincristine sulfate, cyclophosphamide, methotrexate are preferred for chemotherapy.Case: Metastases to all mammary lobes, cervix uteri, neck, skin, gluteal muscles, the oropharyngeal region, and primary vaginal mass were described in spayed bitch, a 10-year old and mixed breed. The clinical examination manifested, fragile and hemorrhagic masses which resemble cauliflower in the vagina, neck, and inguinal region. Furthermore firm and multilobular masses in all mammary lobes, oropharyngeal region, and gluteal muscles of right leg were detected. Firstly, vaginal cytology was performed in order to confirm...(AU)

Clinicopathological evaluation of disseminated metastases of transmissible venereal tumor in a spayed bitch

Background: Although transmissible venereal tumor (TVT, transmissible venereal sarcoma, Sticker’s sarcoma) that affects dogs and other canids can be seen in many countries, it especially emerges in the countries which homeless dog population is very high. Female dogs are more susceptible than males. Transmissible venereal tumor is usually transmitted to genital organs during coitus and occasionally by social behavior such as sniffing and licking. The tumor is generally observed in the posterior part of the vagina. The tumor usually appears in various sizes, in the appearance of cauliflower, red and fragile. Metastases are rarely reported in cases with TVT. Metastases have been detected in lung, liver, tonsils, skin, lymph nodes, muscles, spleen. The diagnosis of transmissible venereal tumor is achived by considering the history of the animal, gross lesions, cytological examination and histopathology. Chemotherapy is frequently used in the treatment of TVT. In addition, radiotherapy, cryosurgery, surgical incision and immunotherapy are rarely applied for treatment. Chemical agents such as doxorubicin, vincristine sulfate, cyclophosphamide, methotrexate are preferred for chemotherapy.Case: Metastases to all mammary lobes, cervix uteri, neck, skin, gluteal muscles, the oropharyngeal region, and primary vaginal mass were described in spayed bitch, a 10-year old and mixed breed. The clinical examination manifested, fragile and hemorrhagic masses which resemble cauliflower in the vagina, neck, and inguinal region. Furthermore firm and multilobular masses in all mammary lobes, oropharyngeal region, and gluteal muscles of right leg were detected. Firstly, vaginal cytology was performed in order to confirm...