Canine Seminomas Research Articles

Histological Typing and Morphological Characterization of Canine Seminomas

The aim of this study was to evaluate seminomas based on cell morphology and histological structures and classify them into subtypes. Thirty canine seminomas were stained with hematoxylin-eosin (HE) and Periodic Acid Schiff (PAS). PAS-positive cases were classified as Classical seminoma (CS) (n:5), while PAS-negative cases were categorized as Spermatocytic seminoma (SS) (n:23). Cases exhibiting both positive and negative staining were evaluated as Spermatocytic/Classical seminoma (n:2). Carcinoma in situ were observed in the cases with Classical seminoma (n:3). The cell morphology and mitotic indices of each case were given in detail. Canine seminomas were classified into three subtypes: intratubular (n:12), diffuse (n:1), and intratubular/diffuse (n:10). As a result, it was thought that SS and CS types could be observed together, and it was observed that the intratubular/diffuse subtype is a third type characterized by its transformation from intratubular to diffuse form.

Testicular tumors in 190 dogs: clinical, macroscopic and histopathological aspects

ABSTRACT: This study aimed to characterize the prevalence and clinical, macroscopic and histopathological aspects of dogs affected by testicular tumors based on biopsy specimens from the Laboratório de Patologia Veterinária of the Universidade Federal de Santa Maria (LPV-UFSM) over 19 years. Parameters regarding the age, size, and breed of the affected dogs were also established. Of all dogs with some type of neoplasm submitted to histopathological analysis at the LPV over these 19 years (n=1,900), 213 (11.2%) had at least one testicular neoplasm. The tissues of 190 dogs (with 220 neoplasms) were available for histological reassessment. The dogs in this study had different types of testicular tumors with relatively similar frequencies. In descending order, the most frequent testicular neoplasms were seminomas (88/220), Leydig (interstitial) cell tumor (LCT; 64/220), Sertoli cell tumor (SCT; 61/220), and mixed germ cell-sex cord stromal tumor (MGSCT) (07/220). Among the dogs of defined breed (119 cases), large breeds had the largest number of cases (50/119), followed by small (47/119) and medium-sized (22/119) breeds. The ages of dogs affected by testicular tumors ranged from 10 months to 18 years. Increased testicular volume was the most common clinical manifestation. Eleven dogs presented information about clinical signs suggestive of hyperestrogenism syndrome (feminization). In seminomas, the diffuse pattern predominated over the intratubular pattern. Two sites (luminal and basal compartments) suggestive of the onset of neoplastic transformations in germ cells were observed in intratubular seminomas. They corroborate the hypothesis that canine seminomas possibly have pathogenesis similar to that observed in human spermatocytic seminomas. The SCTs and LCTs presented high cell morphology variation. SCTs had neoplastic cells organized in five different histological arrangements. As for LCT, solid-diffuse and cystic-vascular histological patterns were the most commonly observed. Through this study, it was possible to establish some of the leading clinical, macroscopic, and histopathological aspects of testicular neoplasms diagnosed over 19 years in the area covered by the LPV-UFSM.

Expression of Protein Gene Product 9.5 and Sal-like Protein 4 in Canine Seminomas

The aim of the present study was to characterize canine classical seminoma (SE) and spermatocytic seminoma (SS) by immunohistochemical expression of gonocytic and spermatogonial cellular markers (c-Kit, placental alkaline phosphatase [PLAP], protein gene product 9.5 [PGP9.5]and Sal-like protein 4 [Sall4]) and histochemically by the periodic acid-Schiff (PAS) reaction. Twenty-five cases of SE and 23 cases of SS were investigated. Two cases of dysgerminoma were also examined. c-Kit was expressed on the cell membrane of 13 of 25 cases of SE (52%) and four of 23 cases of SS (16%). This marker was not expressed in dysgerminoma. PLAP immunoreactivity was observed in the cytoplasm of neoplastic cells of six of 25 cases of SE (24%). PLAP was not expressed in cases of SS and dysgerminoma. All samples of SE, SS and dysgerminoma showed cytoplasmic expression of PGP9.5 and nuclear immunoreactivity for Sall4. There was fine granular cytoplasmic PAS staining in neoplastic cells in five of 25 cases of SE (20%), while all samples of SS and dysgerminoma cases were PAS negative. These findings suggest that it is not possible to differentiate canine SE and SS using these markers. This may be because canine SS may be derived from spermatogonia that can differentiate to spermatocytesand also because cases of canine SE might consist of neoplastic cells that have lost their gonocytic nature. This study was the first to show positive immunoreactivity for Sall4 in canine seminomas and dysgerminomas and expression of PGP9.5 in canine dysgerminomas.

Morphological and immunohistochemical characterisation of seminomas in Norwegian dogs

BackgroundSeminomas in the dog have traditionally been assumed to resemble human spermatocytic seminomas, based on their low malignancy and high occurrence in old individuals. However, recently published studies indicate that canine seminomas can be classified as classical and spermatocytic seminomas in a similar way as in man, and that classical seminomas comprise a substantial proportion of seminomas in the dog. These two factors both contribute to increasing the potential of canine seminoma as a relevant model for human testicular cancer. The aim of the present study was to characterise seminoma in Norwegian dogs using morphology and immunohistochemistry, and determine whether these tumours are comparable with human classical seminoma.MethodsBy applying diagnostic criteria from human pathology, 45 seminomas from the Norwegian Canine Cancer Register were examined histologically with hematoxylin and eosin (HE) and periodic acid-Schiff (PAS) stains. All sections were stained immunohistochemically with antibodies against human placental alkaline phosphatase (PLAP) and the transmembrane receptor c-KIT.ResultsAlthough two of the seminomas showed immunohistochemical staining characteristics indicative of classical seminoma (PLAP+/c-KIT+), all 45 examined seminomas were morphologically consistent with spermatocytic seminoma.ConclusionsThe value of canine seminoma as a model for SE in man remains unclear. Among the 45 investigated tumours from Norwegian dogs, none were classified as classical seminoma based on morphological criteria consistent with human seminomas. Regional or breed differences in the occurrence of classical seminoma in the dog, as well as the lack of uniform diagnostic criteria, might explain the discrepancy between the findings in the current study and the results presented by other authors.

Infiltrating Foxp3+ Regulatory T cells and Histopathological Features in Canine Classical and Spermatocytic Seminomas

In humans, regulatory T (T reg) cells are known to play a critical role in both the regulation of immune homoeostasis and the progression of cancer. However, there is little information about the identification, characterization and the function of T reg cells in canine tumours. We identified T reg cells in 28 canine seminoma samples using a Forkhead box P3 (Foxp3) antibody and investigated the relationship between T reg cell infiltration and histopathological features of classical and spermatocytic seminomas (SE and SS, respectively). The Foxp3 protein showed nuclear immunostaining in infiltrating lymphocytes, and Foxp3+ cells were diffused or focally distributed in seminoma tissues. Foxp3+ cells were frequently present in the SS histotype, in seminomas that showed no evidence of tumour cell invasion into the vessels and in seminomas showing a diffuse growth pattern with three cell types. Neither the SE/SS histotype nor the histopathological features of the tumour correlated with Foxp3+ cell counts. These results indicate that Foxp3+ T reg cells may be associated with a less malignant histological phenotype or may not play a critical role in the immune response of canine seminomas. Moreover, Foxp3+ T reg cells may be associated with SS seminoma, but further studies, involving a larger number of samples, are required to better understand whether these cells play a critical role in the immune response in canine seminomas. This is the first report to demonstrate the characteristics of T reg cell infiltration in canine seminoma.

Immunohistochemical evaluation of lymphocyte types infiltrate into the canine seminomas

BACKGROUNDS: Seminoma is frequently observed in human and canine testes especially in cryptorchids. Canine and human seminomas are typically associated with leukocytic infiltration. OBJECTIVES: We aimed to identify the type of lymphocytes that infiltrate to seminomas. METHODS: Tumor infiltrating lymphocytes were evaluated by immunohistological techniques in 5 dogs with diffuse seminoma. Routine pathological examination of these specimens showed diffuse seminoma with tumor infiltrating lymphocytes in all cases. RESULTS: Lymphocytes in seminomas were predominantly T cytotoxic (CD3+, CD8+) cells. These cells were distributed diffusely, but patchy around the vessels. B-cells were also identified as some rare single cells diffusely scattered in the tumoral parenchyma. CONCLUSIONS: The results of this study showed that the lymphocyte infiltrating cells are mainly T lymphocytes particularly CD8 cytotoxic type. BACKGROUNDS: Seminoma is frequently observed in human and canine testes especially in cryptorchids. Canine and human seminomas are typically associated with leukocytic infiltration. OBJECTIVES: We aimed to identify the type of lymphocytes that infiltrate to seminomas. METHODS: Tumor infiltrating lymphocytes were evaluated by immunohistological techniques in 5 dogs with diffuse seminoma. Routine pathological examination of these specimens showed diffuse seminoma with tumor infiltrating lymphocytes in all cases. RESULTS: Lymphocytes in seminomas were predominantly T cytotoxic (CD3+, CD8+) cells. These cells were distributed diffusely, but patchy around the vessels. B-cells were also identified as some rare single cells diffusely scattered in the tumoral parenchyma. CONCLUSIONS: The results of this study showed that the lymphocyte infiltrating cells are mainly T lymphocytes particularly CD8 cytotoxic type.

Testicular germ cell tumours in dogs are predominantly of spermatocytic seminoma type and are frequently associated with somatic cell tumours

Unlike seminomas in humans, seminomas in animals are not typically sub-classified as classical or spermatocytic types. To compare testicular germ cell tumours (TGCT) in dogs with those of men, archived tissues from 347 cases of canine testicular tumours were morphologically evaluated and characterized using human classification criteria. Histopathological and immunohistological analysis of PLAP, KIT, DAZ and DMRT1 expression revealed that canine seminomas closely resemble human spermatocytic seminomas. In addition, a relatively frequent concomitant presence of somatic cell tumours was noted in canine TGCT. None of the canine TGCT evaluated demonstrated the presence of carcinoma in situ cells, a standard feature of human classical seminomas, suggesting that classical seminomas either do not occur in dogs or are rare in occurrence. Canine spermatocytic seminomas may provide a useful model for this rare human neoplasm.

Canine classical seminoma: a specific malignant type with human classifications is highly correlated with tumor angiogenesis

BackgroundHuman seminoma is classified as classical seminoma (SE) and spermatocytic seminoma (SS). Human SE is known to be more malignant and metastasizing more frequently than SS. Tumor angiogenesis is highly related with tumor progression and metastasis, with microvessel density (MVD) being an important parameter of metastatic potential. Canine seminoma is not yet well-established as SE or SS type including correlation with angiogenesis. We classified canine SE and SS, and then compared them to tumor associated vessels.MethodsTwenty-three cases of canine seminomas (2 intratubular, 9 diffuse, and 12 intratubular/diffuse seminomas showing both intratubular and diffuse patterns) were classified as SE or SS by immunohistochemistry (IHC) using monoclonal antibody against PLAP and by PAS stain. The histopathological data were then compared to see if there was a correlation with SE or SS. Angiogenesis of seminomas were evaluated by immunohistochemical assay using polyclonal antibody against Von Willebrand factor (vWF) and by calculating the means of MVD, vessels area and perimeters using computerized image analysis. Statistical Package for Social Sciences (SPSS) program was used for various statistical analyses.ResultsThe numbers of PLAP+/PAS+ canine SEs were 8/23 (34.8%) and PLAP-/PAS- SSs were 15/23 (61.2%). All SE cases (8/8, 100%) were intratubular/diffuse types. SS types included 2 intratubular (2/15, 13.3%), 9 diffuse (9/15, 60%), and 4 intratubular/diffuse (4/15, 26.7%) types. MVD and vascular parameters in SEs were significantly higher than in SSs, showing the highest value in the intratubular/diffuse type. Seminomas observed with neoplastic cells invasion of vessels presented higher perimeter and area values than seminomas without conformed neoplastic cells invasion.ConclusionIn this study, we demonstrated a positive relationship between canine SE and tumor angiogenesis. Furthermore, we also showed that a tumor cells invasion of vessels were a correlated vascular parameter. Although metastasis of canine seminomas has rarely been reported, our results support that canine SE could have high metastatic potential similar to the human counterpart. Further studies are required to clarify the relationship between canine SE and clinical data with metastatic factors.

Comparative immunohistochemical characterization of canine seminomas and Sertoli cell tumors

Primary testicular tumors are the most common causes of cancer in male dogs. Overall, the majority of canine patients should be cured by testicular surgery. However, tumor markers are not well-known in veterinary medicine. We sought to determine using immunohistochemistry whether the combined human testicular tumor markers (placental alkaline phosphatase, OCT3/4, CD30, alpha-fetoprotein, inhibin-alpha, vimentin, c-KIT, and desmin) are expressed in canine seminomas and Sertoli cell tumors (SCTs). We examined 35 canine testicular tumors, 20 seminomas and 15 SCTs. c-KIT was expressed markedly in canine seminomas. Both inhibin-alpha and vimentin were expressed significantly in canine SCTs. The results of this study demonstrate differences and similarities between tumor marker expression of testicular tumors in dogs and humans. All the main markers in current routine use are discussed as well as potential useful markers for benign and malignant tumors, and tumor progression.

Coelomic mass in a rock dove (Columba livia)

A 15-year-old domestic rock dove (Columbia livia) was presented to the Service de Médecine Zoologique of the Université de Montréal with a 10-day history of lethargy and chronic weight loss of 1-year duration. Ultrasonographic evaluation of the coelomic cavity revealed the presence of effusion and a well-defined mass containing variably sized hypoechoic nodules of unknown origin. Cytologic evaluation of fine-needle aspirates of the mass indicated a malignant epithelial tumor consisting of round cells arranged individually and in clusters, with large nuclei, prominent nucleoli, moderate anisocytosis and anisokaryosis, atypical mitoses, occasional binucleation and multinucleation, and large numbers of ruptured cells. A diagnosis of seminoma was established on histologic sections obtained at necropsy. The cytologic features of this tumor were similar to those of canine seminomas except for the presence of lymphocytes, which were not observed in significant numbers in cytologic smears or histologic sections. Seminoma should be included in the differential diagnosis for a space-occupying mass causing organ displacement within the coelomic cavity of a male bird.

Genomic and expression profiling of human spermatocytic seminomas: pathogenetic implications

Traditionally, germ cell tumours (GCTs) have been classified based on their histological appearance, i.e. the differentiation of lineages. Various subtypes of seminomas have been identified in a number of classification systems, including anaplastic and spermatocytic seminoma. Here we present conclusive evidence that spermatocytic seminomas are GCTs with a separate pathogenesis from seminoma, and should therefore not be considered as a variant of seminoma. Moreover, the different putative animal models, either spontaneous or induced, are discussed in the context of their value as model for human GCTs. The canine seminomas are currently considered as the most informative model for human spermatocytic seminomas.

Correlation of Nuclear Morphometric Features with Animal and Human World Health Organization International Histological Classifications of Canine Spontaneous Seminomas

The aim of this study was to correlate nuclear morphometric features with animal and human World Health Organization International Histological Classifications in canine seminomas. Twenty-three canine seminomas were classified, according to Animal World Health Organization International Histological Classification as intratubular, intratubular with signs of invasion, or diffuse and according to Human World Health Organization International Histological Classification criteria as spermatocytic and typical. The morphonuclear characteristics of tumors were quantitatively evaluated by means of digital cell image analyses of hematoxylin and eosin-stained nuclei. In particular, the mean nuclear area, mean nuclear perimeter, mean nuclear form factor, and their respective standard deviations were calculated. The relationship between the different variables and the tumor histologic types was assessed. On the basis of animal and human classification systems, statistically significant differences were observed only between intratubular seminomas with signs of invasion and the other two types and between spermatocytic and typical seminomas, respectively. In humans, it is well known that typical seminomas are more common and aggressive than spermatocytic ones. In our study, the canine seminomas classified as typical showed significantly larger and more variable nuclear area and perimeter than spermatocytic seminomas. These results support the opinion that most canine seminomas correspond to human spermatocytic seminomas and could explain the benign behavior of canine seminomas, which derive from a more differentiated type of germ cell.

Immunohistochemical Characterization of the Leucocytic Infiltrate Associated with Canine Seminomas

Both canine and human seminomas are typically associated with leucocytic infiltration, the role of which remains poorly understood. In this study, leucocytes infiltrating 10 canine seminomas were characterized. Monoclonal antibodies directed against CD18, CD11a, CD11b, CD11c, CD21, CD3, CD4, CD8 and Major Histocompatibility Complex class I and II (MHC I and MHC II) were also employed. Infiltrating leucocytes were located around vessels, adjacent to the thin fibrous septa between neoplastic lobules, and were also scattered singly amongst neoplastic cells. The more profuse infiltrates often had the appearance of follicular aggregates. Immunohistochemically, all the samples showed generally similar results. Most of the infiltrating cells were positive for CD18 and CD11a. Infiltrating cells were mainly T lymphocytes (CD3 +), particularly of the CD8 + subset. B lymphocytes (CD21 +) were detectable in almost all infiltrates; in the follicular aggregates they were centrally located, whereas T lymphocytes (CD3 +) lined the periphery. CD11c + cells were always more numerous than CD11b + cells, demonstrating that if macrophages and antigen-presenting cells (APCs) were well represented, monocytes and granulocytes were practically absent. Almost all of the infiltrating cells were positive for both MHC I and MHC II antigens and, in nine samples, a large number of neoplastic cells expressing MHC I were detected. A few neoplastic cells expressing MHC II were observed in seven cases. The presence of CD8 + lymphocytes, together with the large number of both infiltrating and neoplastic cells expressing MHC I, suggests that the lymphocytes mediate a cytotoxic reaction against the neoplastic cells. This hypothesis may underlie the favourable prognosis frequently associated with canine seminomas.

Evaluation of Angiogenesis in Canine Seminomas by Quantitative Immunohistochemistry

Angiogenesis, which assists in supplying the nutritional and respiratory needs of proliferating cells, is essential for tumour growth. Angiogenic control is complex, involving a network of cytokines, in particular vascular endothelial growth factor ( VEGF ), a potent endothelial cell mitogen which also stimulates neoplastic cell proliferation. The purpose of this study was to evaluate VEGF expression and microvessel density (number of microvessels per mm2), in canine seminomas. VEGF expression and microvessel density were higher in seminomas than in normal testicular tissue; both parameters were higher in diffuse tumours than in intratubular tumours. These data demonstrate an increase in angiogenesis in the more malignant histological types of seminoma and suggest that both VEGF and microvessel density are useful criteria for evaluating the intrinsic malignancy and growth potential of canine testicular tumours.

Immunohistochemical analysis of cyclins in canine normal testes and testicular tumors.

The expression of cyclins A, D1, D2 and E were examined immunohistochemically in 5 canine normal testes and 31 testicular tumors, including 14 seminomas, 11 Sertoli cell tumors and 6 Leydig cell tumors. In canine normal testes, cyclin A expression was detected in spermatogonia and primary spermatocytes. This suggests that A-type cyclins may play some role in canine spermatogenesis. Cyclin A expression was also observed in 13/14 (92.9%) seminomas and 2/11 (18.2%) Sertoli cell tumors, but no positive reaction was observed in Leydig cell tumors. Parallel examinations for cyclins D1, D2 and E gave negative results in canine normal testes and testicular tumors. High levels of cyclin A expression in canine seminomas indicate that the neoplastic germ cells may be arrested at the spermatogonia and primary spermatocyte stages of differentiation.

Number and size of silver-stained nucleoli (Ag-NOR clusters) in canine seminomas: Correlation with histological features and tumour behaviour

Twenty spontaneous canine seminomas were examined. A statistically significant difference (P < 0.01) was detected in both the mean silver-stained nucleolar area/cell and the mean silver-stained nucleolar number/cell between (1) two intraductal seminomas with no signs of invasion, and (2) three intraductal ones with signs of invasion and 15 diffuse-type seminomas (two of which metastasized 3 and 12 months after surgical treatment). Furthermore, the metastasizing seminomas showed a mean silver-stained nucleolar area/cell which was significantly greater than that observed in all the other seminomas examined (P < 0.01). It is concluded that quantitative studies of nucleolar changes are useful in evaluating the biological behaviour of spontaneous canine seminomas.

The ultrastructure of testicular tumours in the dog: I. Germinal cells and seminomas

The ultrastructure of the cells of normal germinal epithelium of healthy dogs is described and compared with that of tumour cells of 16 canine seminomas. Both normal and tumour cells are characterized by oval nuclei, straight cell borders, relative scarcity of cytoplasmic organelles, and distinct Golgi complex. Intercellular bridges, as seen in normal germinal cells, are not present in seminomas, desmosomes occur in tumour cells only. Two types of tumour cells are described, distinguishable from each other by shape, cytoplasmic density and content of organelles. They are considered to be stages of differentiation of one particular tumour cell. In all cases the cells are markedly distinct from cells of Leydig cell tumours and Sertoli cell tumours.