RESUMEN
BACKGROUND: The ocular side effects of cancer chemotherapeutic drugs are relatively uncommon. Nonetheless, the ocular system has a potentially high sensitivity to toxic substances. This study proposed a framework to assess the effect of vincristine chemotherapy on intraocular pressure, tear protein, and oxidative stress in canines with transmissible venereal tumor (TVT). METHODS: The study group comprised 10 dogs with TVT, whose diagnosis was based on cytology, and all dogs were treated with vincristine for 4 weeks. Each animal was given a complete ophthalmic examination, followed by a standard Schirmer tear test. Before and 20 min after administering vincristine, intraocular pressure (IOP) was measured in the eyes with a noncontact tonometer. At any of the times mentioned, tear samples were collected using the Schirmer test procedure and were subjected to protein analysis-oxidative stress index (OSI), total antioxidant capacity (TAC), total oxidant status (TOS), nitric oxide (NO), and malondialdehyde (MDA) were determined, and standard statistical analysis was applied. RESULTS: No significant differences were found in protein in tears, but mean Pre and Postinjection IOP revealed a significant decrease in the eyes each week. Also, results indicated significant differences in oxidative stress markers: increased OSI, NO, and MDA, and reduced TAC. CONCLUSION: The importance of an increase in oxidative stress levels in the tears of vincristine-treated patients should be taken seriously, as it appears to play a role in the pathogenesis of eye disease. Therefore, during the treatment weeks prior to prescribing vincristine, eye diseases should be evaluated and considered.
Asunto(s)
Oftalmopatías , Tumores Venéreos Veterinarios , Humanos , Animales , Perros , Vincristina/efectos adversos , Presión Intraocular , Tumores Venéreos Veterinarios/tratamiento farmacológico , Tumores Venéreos Veterinarios/metabolismo , Tumores Venéreos Veterinarios/patología , Oftalmopatías/metabolismo , Oftalmopatías/veterinaria , Lágrimas/metabolismo , Estrés OxidativoRESUMEN
Canine transmissible venereal tumor (CTVT) is a tumor that commonly occurs in genital and extragenital sites of both genders. Long interspersed nuclear elements (LINE-1) retrotransposon has a pivotal role in allogenic transfection among uncontrolled dog populations. This study aimed to perform pathomorphological, immunohistochemical, and in situ polymerase chain reaction (PCR) evaluation of CTVT (n = 18) in transfected dogs during chemotherapy. Immunohistochemically, tumor phases were investigated by using specific markers (CD3, CD4, CD8, CD79, and transforming growth factor beta [TGF-ß]), and investigated an amplified specific sequence of TVT LINE-1 retrotransposon by in situ PCR. Polyhedral-shaped neoplastic cells that had large, round, hypo/hyperchromatic nuclei and eosinophilic cytoplasm were detected. All marker results were positive, especially in the early weeks of recovery. CD4 and TGF-ß markers were conspicuously positive at the initial stage. In situ PCR LINE-1 sequence was initially positive in only four cases. It is believed that the CD and TGF-ß markers provide phase identification at tumor initiation and during chemotherapy. It is thought that presence of T and B lymphocytes, which have roles in cellular and humoral immunity, is needed so that regression of the tumor is possible.
Asunto(s)
Biomarcadores de Tumor/análisis , Enfermedades de los Perros/diagnóstico , Elementos de Nucleótido Esparcido Largo , Tumores Venéreos Veterinarios/diagnóstico , Animales , Antígenos de Diferenciación de Linfocitos T/análisis , Antígenos CD79/análisis , Enfermedades de los Perros/metabolismo , Enfermedades de los Perros/terapia , Perros , Femenino , Masculino , Transfección/veterinaria , Tumores Venéreos Veterinarios/metabolismo , Tumores Venéreos Veterinarios/terapiaRESUMEN
Matrix metalloproteases (MMPs) are endogenous proteases that are responsible for degradation of extracellular matrix (ECM) proteins and cell surface antigens. The breakdown of ECM participates in the local invasion and distant metastases of malignant tumors. Canine transmissible venereal tumor (CTVT) is a naturally occurring contagious round cell neoplasm of dogs that affects mainly the external genitalia of both sexes. CTVT generally is a locally invasive tumor, but distant metastases also are common in puppies and immunocompromised dogs. We investigated the immune expressions and activities of MMP-2 and MMP-9 in CTVT. The presence of these enzymes in tumor cells and tissue homogenates was demonstrated by immunohistochemistry and western blotting. We used gelatin substrate zymography to evaluate the activities of MMP-2 and MMP-9 enzymes in tumor homogenates. We found that tumor cells expressed both MMP-2 and MMP-9. Electrophoretic bands corresponding to MMP-9 and MMP-2 were identified in immunoblots and clear bands that corresponded to the active forms of MMP-2 and MMP-9 also were detected in gelatin zymograms. Our study is the first detailed documentation of MMPs in CTVT.
Asunto(s)
Enfermedades de los Perros/inmunología , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Tumores Venéreos Veterinarios/metabolismo , Animales , Enfermedades de los Perros/patología , Perros , Femenino , Inmunohistoquímica/métodos , Masculino , Inhibidor Tisular de Metaloproteinasa-2/metabolismoRESUMEN
Mouse Double Minute-2 (MDM2) is an ubiquitin ligase which is overexpressed or its promoter polymorphism has been reported in different tumours. The objective of this study was to examine the MDM2 protein expression and its promoter polymorphism in some canine tumours. Twenty specimens were collected from 20 dogs with 15 mammary gland carcinomas, 3 lymphomas, 1 transmissible venereal tumour and 1 trichoblastoma. Samples were analysed immunohistochemically using human antibody against MDM2 protein. PCR and DNA sequencing were carried out to identify MDM2 promoter polymorphism. MDM2 gene was expressed in 13 of 20 samples including 11 mammary carcinomas, 1 lymphoma and 1 trichoblastoma. We found 94% homology between canine and human sequences. Four mutations including G169C, A177G, G291T and A177G were identified in different types of breast carcinomas. An extra p53 response element was found in a mixed mammary carcinoma.
Asunto(s)
Enfermedades de los Perros/metabolismo , Regulación Neoplásica de la Expresión Génica/fisiología , Polimorfismo Genético , Regiones Promotoras Genéticas/genética , Proteínas Proto-Oncogénicas c-mdm2/metabolismo , Animales , Carcinoma/genética , Carcinoma/metabolismo , Carcinoma/veterinaria , Enfermedades de los Perros/genética , Perros , Linfoma/genética , Linfoma/metabolismo , Linfoma/veterinaria , Proteínas Proto-Oncogénicas c-mdm2/genética , Tumores Venéreos Veterinarios/genética , Tumores Venéreos Veterinarios/metabolismoRESUMEN
BACKGROUND: Chemokines play multiple roles in the development and progression in a variety of tumors. Chemokine (C-X-C motif) ligand 7 (CXCL7) has been found associated with pro-inflammatory responses, but its role in cancer growth remains unclear. Our previous study showed that R phase tumor infiltrating lymphocytes (TILs) produced large amounts of interleukin (IL)-6 which antagonized transforming growth factor (TGF)-ß derived from CTVT to diminish the immune-suppressive microenvironment. Now we intend to determine the expression pattern of CXCL7 and the role of IL-6/TGF-ß in CXCL7 induction during spontaneous progressive (P) and regressive (R) phases in canine transmissible venereal tumor (CTVT). RESULTS: We have demonstrated that CXCL7 expressed at high level in P phase and down-regulated in R phase by western blot and real-time PCR. This suggested that CXCL7 expression was negatively correlated with the tumor growth. Co-culturing TILs with CTVT cells was found to reduce CXCL7 expression, while adding IL-6 blocking antibody reversed it. Moreover, in P phase CTVT, while IL-1ß and TGF-ß had no obvious effect on CXCL7 expression, IL-6 was found significantly to reduce CXCL7 expression in a dose-dependent manner. The mRNA expression results of CXCL7 receptor, CXCR2, further confirmed the effects of IL-6 concentration on the CXCL7 expression. CONCLUSION: CXCL7 overexpression might be associated with the progressive growth of CTVT. The results shown here also suggest the role of CXCL7 in cancer development and the potential as the anti-cancer therapeutic target.
Asunto(s)
Quimiocinas CXC/metabolismo , Enfermedades de los Perros/metabolismo , Regulación Neoplásica de la Expresión Génica/fisiología , Tumores Venéreos Veterinarios/metabolismo , Animales , Células Cultivadas , Quimiocinas CXC/genética , ADN Complementario/genética , ADN Complementario/metabolismo , Perros , Interleucina-6/genética , Interleucina-6/metabolismo , Masculino , ARN Mensajero/genética , ARN Mensajero/metabolismo , Tumores Venéreos Veterinarios/genéticaRESUMEN
Cancer is generally defined as uncontrollable growth of cells caused by genetic aberrations and/or environmental factors. Yet contagious cancers also occur. The recent emergence of a contagious cancer in Tasmanian devils has reignited interest in transmissible cancers. Two naturally occurring transmissible cancers are known: devil facial tumour disease and canine transmissible venereal tumour. Both cancers evolved once and have then been transmitted from one individual to another as clonal cell lines. The dog cancer is ancient; having evolved more than 6,000 years ago, while the devil disease was first seen in 1996. In this review I will compare and contrast the two diseases focusing on the life histories of the clonal cell lines, their evolutionary trajectories and the mechanisms by which they have achieved immune tolerance. A greater understanding of these contagious cancers will provide unique insights into the role of the immune system in shaping tumour evolution and may uncover novel approaches for treating human cancer.
Asunto(s)
Neoplasias/inmunología , Animales , Perros , Neoplasias Faciales/inmunología , Neoplasias Faciales/metabolismo , Humanos , Complejo Mayor de Histocompatibilidad/genética , Complejo Mayor de Histocompatibilidad/fisiología , Marsupiales , Neoplasias/metabolismo , Tasmania , Tumores Venéreos Veterinarios/inmunología , Tumores Venéreos Veterinarios/metabolismoRESUMEN
Canine transmissible venereal tumor (CTVT) is an infectious cell line circulating in many feral dog populations. It originated once, about 10,000 years ago. Phylogenetic analyses of mitochondrial sequences from dogs, wolves, and a geographically diverse collection of CTVT samples indicate that the cancer has periodically acquired mitochondria from its host. We suggest that this may be because the cancer's own mitochondria have a tendency to degenerate, due to high mutation rates and relaxed selection, resulting in host mitochondria being more fit.
Asunto(s)
Enfermedades de los Perros/genética , Genoma Mitocondrial , Mitocondrias , Tumores Venéreos Veterinarios/genética , Tumores Venéreos Veterinarios/metabolismo , Animales , Coyotes/genética , Enfermedades de los Perros/metabolismo , Enfermedades de los Perros/patología , Perros/genética , Transferencia de Gen Horizontal , Mitocondrias/genética , Mitocondrias/metabolismo , Filogenia , Polimorfismo Genético , Selección Genética , Análisis de Secuencia de ADN , Tumores Venéreos Veterinarios/patología , Lobos/genéticaRESUMEN
Many tumors down-regulate major histocompatibility complex (MHC) antigen expression to evade host immune surveillance. However, there are very few in vivo models to study MHC antigen expression during tumor spontaneous regression. In addition, the roles of transforming growth factor betal (TGF-beta1), interferon gamma (IFN-gamma), and interleukin (IL)-6 in modulating MHC antigen expression are ill understood. We previously reported that tumor infiltrating lymphocyte (TIL)-derived IL-6 inhibits TGF-beta1 and restores natural killing (NK) activity. Using an in vivo canine-transmissible venereal tumor (CTVT) tumor model, we presently assessed IL-6 and TGF-beta involvement associated with the MHC antigen expression that is commonly suppressed in cancers. IL-6, IFN-gamma, and TGF-beta1, closely interacted with each other and modulated MHC antigen expression. In the presence of tumor-derived TGF-beta1, host IFN-gamma from TIL was not active and, therefore, there was low expression of MHC antigen during tumor progression. TGF-beta1-neutralizing antibody restored IFN-gamma-activated MHC antigen expression on tumor cells. The addition of exogenous IL-6 that has potent anti-TGF-beta1 activity restored IFN-gamma activity and promoted MHC antigen expression. IFN-gamma and IL-6 in combination acted synergistically to enhance the expression of MHC antigen. Thus, the three cytokines, IL-6, TGF-beta1, and IFN-gamma, closely interacted to modulate the MHC antigen expression. Furthermore, transcription factors, including STAT-1, STAT-3, IRF-1, NF-kappaB, and CREB, were significantly elevated after IL-6 and IFN-gamma treatment. We conclude that the host IL-6 derived from TIL works in combination with host IFN-gamma to enhance MHC molecule expression formerly inhibited by TGF-beta1, driving the tumor toward regression. It is suggested that the treatment of cancer cells that constitutively secrete TGF-beta1 should incorporate anti-TGF-beta activity. The findings in this in vivo tumor regression model have potential applications in cancer immunotherapy.
Asunto(s)
Enfermedades de los Perros/inmunología , Antígenos de Histocompatibilidad Clase II/biosíntesis , Antígenos de Histocompatibilidad Clase I/biosíntesis , Interferón gamma/inmunología , Interleucina-6/inmunología , Regresión Neoplásica Espontánea , Factor de Crecimiento Transformador beta1/metabolismo , Tumores Venéreos Veterinarios/inmunología , Animales , Enfermedades de los Perros/metabolismo , Enfermedades de los Perros/patología , Perros , Femenino , Células Asesinas Naturales/inmunología , Linfocitos Infiltrantes de Tumor/inmunología , Masculino , Unión Proteica , Tumores Venéreos Veterinarios/metabolismo , Tumores Venéreos Veterinarios/patologíaRESUMEN
Mast cells are immune cells that are involved mainly in type 1 hypersensitivity reactions, and they have been implicated in tumour angiogenesis. In this study we assessed the presence of mast cell numbers and microvessel density during the progression and regression stages of natural spontaneous canine transmissible venereal tumours (CTVT). Mast cells were demonstrated by histochemical staining with toluidine blue, alcian blue and safranin O. Microvessel counts were demonstrated by immunohistochemical labelling with an antibody against the endothelial cell marker factor VIII. Mitotic cells, apoptotic cells and tumour infiltrating lymphocytes were counted from haematoxylin-eosin-stained sections. Tumour fibrosis was evaluated on Masson's trichome-stained sections. The results showed that progressing tumours had significantly higher mast cell counts and microvessel counts at the invasive edges of the tumours than did regressing tumours. In both the progressing and regressing tumours, microvessel counts were significantly positively correlated with mast cell counts. Regressing tumours had significantly higher mast cell counts of the whole tumour than progressing tumours. The results also showed that progressing tumours had significantly higher mitotic rate than regressing tumours, and fibrosis and apoptosis were significantly higher in regressing tumours than progressing tumours. There were no significant differences between the biochemical and haematological values of dogs with progressing and regressing tumours. These results suggests that mast cells play a role in CTVT progression probably by promoting vascularization at the invasion front during the progression phase, and that mast cell count could be used as one of the histological factors to indicate growth stage of CTVT.
Asunto(s)
Enfermedades de los Perros/metabolismo , Enfermedades de los Perros/patología , Mastocitos/citología , Invasividad Neoplásica/patología , Regresión Neoplásica Espontánea/patología , Tumores Venéreos Veterinarios/metabolismo , Tumores Venéreos Veterinarios/patología , Animales , Apoptosis , Recuento de Células/veterinaria , Perros , Femenino , Inmunohistoquímica/métodos , Inmunohistoquímica/veterinaria , Linfocitos Infiltrantes de Tumor/patología , Masculino , Índice Mitótico/veterinaria , Estadificación de Neoplasias , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Neovascularización Patológica/veterinariaRESUMEN
One of the most frequent canine neoplasms is the transmissible venereal tumor (TVT), which affects the male and the female genital tract. The objective of this study was to determine (immunohistochemically) estrogen receptor (ER-alpha) expression in vaginal tissue of healthy bitches and in the vaginal and neoplastic tissues of TVT-affected bitches. Fifty-eight bitches were divided into two groups: tumor group (TVT) and control group (healthy). Canine estrous cycle stages were determined by means of exfoliative vaginal cytology, hormone assays, and macroscopic appearance of ovaries. Samples from vaginal and neoplastic tissues were obtained under general anesthesia, fixed in 10% buffered formaldehyde, embedded in paraffin, and sectioned. Anestrus, proestrus and estrus control females had higher ER-alpha expression than diestrus bitches. Within the tumor group, diestrus bitches had significantly higher ER-alpha expression. Although some samples had expression in the endothelium of blood vessels, no ER-alpha expression was observed in neoplastic tissues. In conclusion, vaginal tissue of tumor and control bitches, under different distinct steroid influences, had different ER-alpha expression, whereas ER-alpha expression was not present in neoplastic tissues.
Asunto(s)
Enfermedades de los Perros/metabolismo , Perros/metabolismo , Estradiol/sangre , Receptor alfa de Estrógeno/metabolismo , Progesterona/sangre , Neoplasias Vaginales/veterinaria , Tumores Venéreos Veterinarios/metabolismo , Animales , Biopsia , Enfermedades de los Perros/sangre , Enfermedades de los Perros/patología , Enfermedades de los Perros/cirugía , Perros/sangre , Receptor alfa de Estrógeno/sangre , Ciclo Estral , Femenino , Inmunohistoquímica/veterinaria , Estadísticas no Paramétricas , Neoplasias Vaginales/sangre , Neoplasias Vaginales/patología , Neoplasias Vaginales/cirugía , Tumores Venéreos Veterinarios/sangre , Tumores Venéreos Veterinarios/patología , Tumores Venéreos Veterinarios/cirugíaRESUMEN
Stromal cells and extracellular matrix (ECM) components are important for tumour cell behaviour. Little is known about the role of stromal cells and ECM components in the progression and regression of spontaneous canine transmissible venereal tumour (CTVT). In this study, the stromal cell type was determined by immunohistochemical labelling with antibodies to desmin, vimentin and alpha-smooth muscle actin (alpha-SMA) during the progressive and regressive stages of spontaneous CTVT. The distribution of ECM components tenascin-C, chondroitin sulphate and versican were determined immunohistochemically, and hyaluronan distribution was determined using a biotinylated protein complex with specific affinity for hyaluronan. Stromal cells of tumours in both the progressive and regressive stage were positive for vimentin and negative for desmin. The number of stromal cells expressing alpha-SMA was significantly higher (P=0.001) in regressing tumours, than progressing tumours. These results suggest that the modulation of stromal cells that occurs during the regression of CTVT is similar to that occurring during wound healing. Tenascin-C was weakly expressed in the stroma of tumours in the progressive stage and in regions of the regressing tumours with tumour infiltrating lymphocytes (TILs), but intensely expressed in the stroma of tumours in late regressive stage. In addition, tenascin-C was also expressed in the cytoplasm of some tumour cells in the late regressive stage. A strong stromal tenascin-C intensity was significantly associated with regressing tumours (P=0.001). Strong stromal hyaluronan intensity and a high proportion of hyaluronan-positive tumour cells were significantly associated with progressing tumours (P=0.001). This suggests that hyaluronan is involved in the growth of the tumour. There was no significant difference in the expression of chondroitin sulphate and versican in progressing and regressing tumours.
Asunto(s)
Enfermedades de los Perros/metabolismo , Enfermedades de los Perros/patología , Tumores Venéreos Veterinarios/metabolismo , Tumores Venéreos Veterinarios/patología , Actinas/metabolismo , Animales , Proteoglicanos Tipo Condroitín Sulfato/metabolismo , Sulfatos de Condroitina/metabolismo , Desmina/metabolismo , Perros , Matriz Extracelular/metabolismo , Matriz Extracelular/patología , Femenino , Ácido Hialurónico/metabolismo , Inmunohistoquímica , Lectinas Tipo C , Linfocitos Infiltrantes de Tumor/patología , Masculino , Regresión Neoplásica Espontánea/patología , Regresión Neoplásica Espontánea/fisiopatología , Células del Estroma/metabolismo , Células del Estroma/patología , Tenascina/metabolismo , Versicanos , Vimentina/metabolismoRESUMEN
IL-6 is a multifunctional cytokine that regulates cell growth, differentiation, and cell survival. Many tumor cells produce TGF-beta1, which allows them to evade CTL-mediated immune responses. IL-6 antagonizes TGF-beta1 inhibition of CD3 cell activation. However, whether IL-6 restores NK activity, which also is suppressed by TGF-beta1, is not known. We used canine transmissible venereal tumor (CTVT), which produces TGF-beta1, as a model to determine whether IL-6 restores lymphokine-activated killer (LAK) activity. During the progression phase, CTVT cells stop expressing MHC molecules. During the regression phase, the number of surface MHC molecules increases dramatically on about one-third of tumor cells. Tumor cells that stop expressing MHC should be targeted by NK cells. In this study, we found that TGF-beta1 secreted by CTVT cells suppressed LAK cytotoxicity. Interestingly, tumor-infiltrating lymphocytes (TIL) isolated from regressing CTVT secrete high concentrations of IL-6 and antagonize the anti-LAK activity of tumor cell TGF-beta1. TIL also produce IL-6 during progression phase, but the concentration is too low to block the anti-LAK activity of TGF-beta1. There is probably a threshold concentration of IL-6 needed to reverse TGF-beta1-inhibited LAK activity. In addition, in the absence of TGF-beta1, IL-6 derived from TIL does not promote the activity of LAK. This new mechanism, in which TIL manufacture high concentrations of IL-6 to block tumor TGF-beta1 anti-LAK activity, has potential applications in cancer immunotherapy and tumor prognosis.
Asunto(s)
Citotoxicidad Inmunológica/inmunología , Interleucina-6/metabolismo , Células Asesinas Activadas por Linfocinas/inmunología , Linfocitos Infiltrantes de Tumor/inmunología , Proteínas de Neoplasias/antagonistas & inhibidores , Factor de Crecimiento Transformador beta/antagonistas & inhibidores , Tumores Venéreos Veterinarios/inmunología , Tumores Venéreos Veterinarios/terapia , Adyuvantes Inmunológicos/metabolismo , Adyuvantes Inmunológicos/fisiología , Animales , Sistema Libre de Células/inmunología , Sistema Libre de Células/metabolismo , Técnicas de Cocultivo , Progresión de la Enfermedad , Perros , Femenino , Interleucina-6/biosíntesis , Interleucina-6/farmacología , Interleucina-6/fisiología , Líquido Intracelular/inmunología , Líquido Intracelular/metabolismo , Células Asesinas Activadas por Linfocinas/metabolismo , Subgrupos Linfocitarios/patología , Linfocitos Infiltrantes de Tumor/metabolismo , Linfocitos Infiltrantes de Tumor/patología , Masculino , Monocitos/patología , Regresión Neoplásica Espontánea/inmunología , ARN Mensajero/biosíntesis , Factores Supresores Inmunológicos/antagonistas & inhibidores , Factores Supresores Inmunológicos/metabolismo , Factores Supresores Inmunológicos/fisiología , Factor de Crecimiento Transformador beta/biosíntesis , Factor de Crecimiento Transformador beta/genética , Factor de Crecimiento Transformador beta/fisiología , Factor de Crecimiento Transformador beta1 , Células Tumorales Cultivadas , Tumores Venéreos Veterinarios/metabolismoRESUMEN
Canine transmissible venereal tumor (CTVT) can be allo-transplanted across major histocompatibility complex barriers. The expression of MHC molecules is usually low in the progression (P) stage and then greatly increases during tumor regression (R). We investigated the effects of tumor infiltrating lymphocytes (TIL) on the expression of MHC molecules of CTVT cells. Isolated, viable CTVT cells were inoculated at each of 12 sites (1 x 10(8) CTVT cells per site) on the back of six, mixed-breed dogs. Tumor masses were collected every 2-3 weeks and prepared for histopathologic, immunocytochemistry, flow cytometry and immunoblotting studies. The level of MHC expression on tumor cells from different stages of growth was measured. Initially, expression of MHC I and II molecules in P phase CTVT was low. Twelve weeks post-inoculation (PI), expression increased dramatically and it continued to increase during R phase. Tumor growth slowed after 12 weeks PI and tumors entered R phase around 17 weeks PI. We hypothesize that CTVT evades host immunosurveillance and grows progressively for 12 weeks, when it becomes vulnerable and subject to the host's anti-tumor immune responses. We further demonstrated that R phase, but not P phase, TIL were closely associated with the over-expression of MHC I and II molecules by CTVT cells. The number and proportion of TIL were higher in R phase tumors. Supernatants, from R phase co-cultures (CTVT+TIL) and TIL only, promoted MHC I and II expression on P phase CTVT cells. After culturing alone for 1 month, expression of MHC classes I and II molecules in R phase CTVT cells decreased to the level of P phase CTVT cells. However, the above-mentioned supernatants restored their expression of MHC I and II molecules. In contrast, supernatants from P phase TIL or CTVT cells increased expression slightly or had no effect. Therefore, TIL, not CTVT cells, produce the effective substance (s) to promote the expression of MHC molecules by the tumor cells. Heat treated supernatant was unable to promote the expression of MHC I and II molecules by CTVT cells. In conclusion, TIL isolated from R phase CTVT secreted a heat-sensitive, soluble substance(s) that triggered over-expression of MHC I and II after 12 weeks PI. This caused the tumor to enter R phase and helped stop CTVT growth. Our findings will facilitate the understanding and further investigation of the mechanisms that initiate host immune surveillance against tumors.
Asunto(s)
Enfermedades de los Perros/inmunología , Regulación de la Expresión Génica/inmunología , Antígenos de Histocompatibilidad Clase II/inmunología , Antígenos de Histocompatibilidad Clase I/inmunología , Linfocitos Infiltrantes de Tumor/inmunología , Tumores Venéreos Veterinarios/inmunología , Animales , Western Blotting/veterinaria , Enfermedades de los Perros/metabolismo , Perros , Femenino , Citometría de Flujo/veterinaria , Antígenos de Histocompatibilidad Clase I/biosíntesis , Antígenos de Histocompatibilidad Clase II/biosíntesis , Inmunohistoquímica/veterinaria , Cinética , Prueba de Cultivo Mixto de Linfocitos/veterinaria , Linfocitos Infiltrantes de Tumor/metabolismo , Masculino , Tumores Venéreos Veterinarios/metabolismoRESUMEN
The collective immunohistochemical expression of human lysozyme, human alpha-1-antitrypsin, human CD3 antigen, calf vimentin, human keratins, human lambda light chains,canine immunoglobulins IgG, IgM, and bovine protein S-100 has been analyzed on formalin-fixed, paraffin-embedded tissue sections of 25 spontaneous canine transmissible venereal tumors (CTVT) from both genital and extragenital locations using the avidin-biotin-peroxidase complex technique. Lysozyme immunoreactivity was detected in 10/25 CTVT, alpha-1-antitrypsin in 14/25 CTVT, and vimentin in 25/25 CTVT. All CTVT cells were negative to keratins 5 + 8 of the Moll catalogue (RCK-102), S-100 protein, lambda light-chain immunoglobulins, IgG, IgM, and CD3 antigen. The intratumoral T-and B-lymphocyte infiltrate was differentiated using CD3 antigen, lambda light-chain immunoglobulins, IgG, and IgM, and this technique could be useful to evaluate the regressive or progressive growth stage of venereal tumors. Our findings support the hypothesis of a histiocytic immunophenotype for CTVT, and these staining techniques could be used in the differential diagnosis with lymphomas.
Asunto(s)
Enfermedades de los Perros/patología , Tumores Venéreos Veterinarios/química , Tumores Venéreos Veterinarios/patología , Animales , Complejo CD3/análisis , Complejo CD3/metabolismo , Bovinos , Diagnóstico Diferencial , Enfermedades de los Perros/metabolismo , Perros , Femenino , Humanos , Inmunoglobulina G/análisis , Inmunoglobulina G/metabolismo , Inmunoglobulina M/análisis , Inmunoglobulina M/metabolismo , Inmunohistoquímica/métodos , Queratinas/análisis , Queratinas/metabolismo , Masculino , Muramidasa/análisis , Muramidasa/metabolismo , Proteínas S100/análisis , Proteínas S100/metabolismo , Tumores Venéreos Veterinarios/metabolismo , Vimentina/análisis , Vimentina/metabolismo , alfa 1-Antitripsina/análisis , alfa 1-Antitripsina/metabolismoRESUMEN
Transmissible venereal tumours (Sticker) of 28 dogs were investigated retrospectively. Data of infection, clinical development, pathological and histological findings were evaluated. The majority of tumours was investigated with regard to their ability to produce cytokeratin in their cytoplasm. With the exception of one dog all animals had been in an African or Southern European country some weeks before the tumour was recognized for the first time. Multiple growth was a common feature of the transmissible venereal tumour; they showed low tendency to infiltrate the surrounding tissues. No case of spontaneous regression was recorded. Metastases to the regional lymph nodes and reinfection were observed in one dog each, relapses in six dogs. In no case the tumour was the immediate cause of death. In the cytoplasm of the transmissible venereal tumours no cytokeratin could be detected.