Immunocytochemical characterization of primary cell culture in canine transmissible venereal tumor

Abstract

RESUMO.-[Caracterização imunocitoquímica de culturas primárias de tumor venéreo transmissível canino.]Os anticorpos anti-vimentina, anti-lisozima, anti-alfa 1 antitripsina, anti-CD3 e anti-CD79α foram empregados para a caracterização de culturas primárias de tumor venéreo transmissível canino (TVT). Amostras para cultura primá-ria e imuno-histoquímica foram coletadas de oito cães com diagnóstico clínico e citológico de TVT. Para validar o resultado inmunocitoquímico nas culturas de TVT foi realizada a contagem de cromossomos. Para a análise estatística o teste de Mann-Whitney foi empregado a um nível de significância de p<0.05. As culturas e os tecidos de TVT apresentaram intensa reatividade para vimentina, moderada a leve para Lisozima, moderada para alfa-antitripsina e não houve marcação para CD3 e CD79α. Immunochemistry with anti-vimentin, anti-lysozyme, anti-alpha 1 antitrypsin, anti-CD3 and anti-CD79α antibodies has been used for characterization of primary cell culture in the transmissible venereal tumor (TVT). Samples for primary cell culture and immunohistochemistry assays were taken from eight dogs with cytological and clinical diagnosis of TVT. To validate the immunochemical results in the primary cell culture of TVT, a chromosome count was performed. For the statistical analysis, the Mann-Whitney test with p<0.05 was used. TVT tissues and culture cells showed intense anti-vimentin immunoreactivity, lightly to moderate immunoreactivity for anti-lysozyme, and mild for anti-alpha-antitrypsin. No marking was achieved for CD3 and CD79α. All culture cells showed chromosomes variable number of 56 to 68. This is the first report on the use of immunocytochemical characterization in cell culture of TVT. Significant statistic difference between immunochemistry in tissue and culture cell was not established, what suggests that the use of this technique may provide greater certainty for the confirmation of tumors in the primary culture. This fact is particularly important because in vitro culture of tumor tissues has been increasingly used to provide quick access to drug efficacy and presents relevant information to identify potential response to anticancer medicine; so it is possible to understand the behavior of the tumor. Immunocytochemical characterization of primary cell culture in transmissible venereal tumor o uso da immunocitoquímica para a caracterização de culturas de TVT. Assim, e devido ao fato de se observar semelhança entre a imunomarcação em células e tecidos, sugere-se que o uso desta técnica possa auxiliar na confirmação de culturas primárias do tumor, fato muito importante porque a utilização da cultura do tumor pode permitir o acesso a informação relevante sobre resposta potencial a um tratamento e conhecimento do comportamento biológico do tumor.TERMOS DE INDEXAÇÃO: Tumor venéreo transmissível canino, alfa 1 antitripsina, citogenética, lisozima, vimentina.

Evaluation of a canine transmissible venereal tumour cell line with tumour immunity capacity but without tumorigenic property

“…In addition, the primary CTVT cell line was inoculated intravaginally to test its tumorigenic potential in the present study, which was methodologically unlike Chien et al . (9) who implanted the cells subcutaneously into the backs of the dogs. However, no tumour growth was observed over a one-year period of evaluation, which was a result consistent with observations made by Adams et al .…”

Does the tumour microenvironment alter tumorigenesis and clinical response in transmissible venereal tumour in dogs?

“…The samples were stored in saline and phosphate solution (PBS) pH 7.4 and in RNA Later (Qiagen, Venlo, Limburg, the Netherlands) in the Department of Pathology, Faculty of Veterinary Medicine and Animal Science—UNESP ‐ Botucatu until processing of the material. To verify that tissues and culture cells coming from as pertaining to CTVT, samples were subjected to immunocytochemistry, as well as analyses of chromosome numbers in the Animal Genetics Laboratory of the Institute of Biosciences UNESP—Botucatu …”

Do alterations in gene expressions influence tumorigenesis in the transmissible venereal tumor in dogs?

“…Tissue samples and insulations of CTVT cultures had the number of chromosomes analyzed at the Animal Genetics Laboratory of the Biosciences Institute (FLÓREZ et al, 2016).…”

“…Insulations of CTVT cultures from the tumor collections (Table 1) were performed according to the protocol described by HSIAO et al (2008) and FLÓREZ et al (2016). For this purpose, tumor fragments were placed in PBS pH 7.4 and taken to the In vitro Fertilization and Cell Culture Laboratory of the Animal Reproduction and Veterinary Radiology Department of Veterinary Hospital.…”

“…The Canine transmissible venereal tumor (CTVT) arouses great interest in oncology researchers due to several characteristics, among them its controversial origin (MUKARATIRWA et al, 2003;FLÓREZ et al, 2016;DUZANSKI et al, 2017), questionable spontaneous regression RESUMO: O tumor venéreo transmissível canino (TVTC) se trata de uma neoplasia transmissível, que se propaga naturalmente entre os cães pela transferência halogênica de células tumorais, principalmente, durante o coito. É a mais antiga linhagem tumoral conhecida na natureza e relatos sobre mutações gênicas vêm sendo ampliadas.…”