Canine pancreatic endocrine tumors: immunohistochemical analysis of hormone content and amyloid

Thirty-one primary canine pancreatic endocrine tumors and their metastases were studied histologically and immunohistochemically for the presence of insulin, glucagon, somatostatin, pancreatic polypeptide (PP), gastrin, and adrenocorticotrophic hormone (ACTH). Tumors were also evaluated for the presence of amyloid. The cytoarchitectural pattern of 25 of 31 primary tumors was predominantly solid, whereas three tumors were mostly glandular, two were unclassified, and one had a gyriform pattern. Cells with insulin immunoreactivity were found in 30 of 31 tumors and were found in all cases in which there was clinical evidence of inappropriate insulin secretion. Insulin was the only hormone demonstrable in three of the 30 tumors, but cells immunoreactive for other hormones were also present in various combinations in most tumors [i.e., glucagon (13 of 30), somatostatin (17 of 30), PP (25 of 30), and gastrin (2 of 30)]. One tumor contained only cells with glucagon and PP immunoreactivity. Amyloid was found in ten of 31 primary tumors but was not detected in metastases. Cells with insulin immunoreactivity were the only cell type consistently present in tumors containing amyloid. Amyloid deposits did not immunoreact with any of the antisera. Seventeen of 31 dogs had metastasis of the pancreatic endocrine tumor to regional lymph nodes, liver, or both. All metastases available for study (15 of 17) contained cells with insulin immunoreactivity and some contained cells with PP or somatostatin immunoreactivity. No statistically significant (P greater than 0.05) differences in tendency to metastasize were found when pancreatic endocrine tumors were compared by region of origin, cytoarchitectural pattern, presence of amyloid, or by number of hormones contained within the tumor.     

Immunocytochemistry of canine thyroid tumors.

Immunocytochemical studies using the peroxidase-antiperoxidase method with commercial antibodies against thyroglobulin, calcitonin, calcitonin gene-related peptide (CGRP), neuron specific enolase (NSE), somatostatin, and neurotensin were performed on 38 Bouin-fixed, paraffin-embedded canine thyroid tumors obtained from necropsy and surgical files from 2 Ecoles Nationales Vétérinaires (Alfort and Nantes, France) and from the Laboratoire d'Histo-Cytopathologie Vétérinaire, Maisons-Alfort (France). The tumors consisted of two follicular adenomas, nine follicular carcinomas, nine solid carcinomas, 12 follicular-compact-cellular carcinomas, and six C-cell carcinomas. All 32 follicular-cell tumors were stained positively for thyroglobulin, half of them had weak to moderate positive immunoreactivity for NSE, and all histologic patterns were represented. They had no immunoreactivity for somatostatin or neurotensin. Four C-cell carcinomas had a solid alveolar pattern, while two had a pseudo follicular pattern characterized by uneven, often coalescent, pseudo-follicular formations with a multilayered epithelium surrounding a cavity that often contained red blood cells. Four C-cell carcinomas had uneven immunoreactivity for calcitonin, while all six were positive for CGRP or NSE. Immunoreactivity for CGRP was stronger or more widespread than positivity for calcitonin when both occurred in the same tumor. Some cells of three C-cell carcinomas had positive immunoreactivity for somatostatin. No immunoreactivity for neurotensin was detected. Seven tumors of follicular cell origin contained a few cells positive for calcitonin or CGRP, while three C-cell carcinomas had a few cells positive for thyroglobulin. These tumors were considered to contain entrapped remnants of normal thyroid tissue rather than being dual hormone producing tumors.     

A malar granular cell tumor in a djungarian hamster

'Granular cell' tumor observed in the malar subcutis of a Djungarian hamster was examined to determine its cellular origin. Histologically, the tumor consisted of a solid growth of oval or spindle-shaped large cells with abundant cytoplasm filled with eosinophilic granules that were periodic acid-Schiff-positive and diastase-resistant. Immunohistochemically, the tumor cells were positive to anti-vimentin and anti-desmin antibodies and a few cells showed positivity to anti-actin antibody as well. They did not react to myoglobin, S-100 protein, and glial fibrillary acidic protein (GFAP). Electron microscopic studies revealed that the tumor cells had pinocytotic vesicles, dense plaque and microfilaments. The first granular cell variant of myogenic tumor reported here in Djungarian hamsters was differentiated from granular cell tumor of Schwann cell origin.     

Characterization of uterine granular cell tumors in B6C3F1 mice: a histomorphologic, immunohistochemical, and ultrastructural study

The granular cell tumor is most often a benign neoplasm of uncertain origin. Four uterine granular cell tumors in control and treated female B6C3F1 mice were identified in chronic studies at the National Toxicology Program. Two tumors occurred in untreated control animals and 2 in treated animals receiving different compounds. Tissue sections were evaluated histologically and stained with hematoxylin and eosin, periodic acid-Schiff with diastase resistance, Masson's trichrome, toluidine blue, phosphotungstic acid-hematoxylin, and stained immunohistochemically with a panel of antibodies to muscle (desmin, alpha smooth muscle actin), neural (S-100, neuron specific enolase), epithelial (wide-spectrum cytokeratin), and macrophage (F4/80) markers. The main histomorphologic feature of tumor cells was the presence of abundant cytoplasmic eosinophilic granules that stained positive for periodic acid-Schiff with diastase resistance. Tumors varied in appearance and were comprised of sheets and nests of round to polygonal cells with distinct borders. Nuclei were hyperchromatic, pleomorphic, and centrally to eccentrically located and often contained single nucleoli. Occasional multinucleated giant cells were observed. Tumors were pale pink and homogeneous with trichrome stain and negative with toluidine blue. Three tumors had positive to weakly positive immunoreactivity for desmin, and 1 was positive for alpha smooth muscle actin. Expression of S-100, wide-spectrum cytokeratin, and neuron-specific enolase was negative for all tumors. Ultrastructurally, prominent electron-dense cytoplasmic granules were abundant and contained secondary lysosomes with heterogeneous lysosomal contents. The characteristics of these uterine granular cell tumors were suggestive of a myogenic origin.     

Canine intravascular lymphoma with overt leukemia

A 6-year-old spayed Labrador Retriever Mix dog was evaluated for a 2-week history of progressive generalized weakness and reluctance to stand. Physical examination revealed severe weakness with obtunded mentation, head tilt, bilateral nystagmus, and decreased vision. CBC findings included mild nonregenerative anemia, marked thrombocytopenia, and a few atypical mononuclear cells on the blood film. The cells were 15-30 μm in diameter and had round to oval to reniform centrally placed nuclei with stippled chromatin, prominent nucleoli, and abundant basophilic cytoplasm with numerous discrete vacuoles and, occasionally, small azurophilic granules. Similar cells were found in bone marrow. On histologic examination of tissues collected at necropsy, neoplastic cells were detected in bone marrow, hepatic sinusoids, cerebral and meningeal vessels, and in capillaries of the heart, renal interstitium, small intestinal submucosa, and muscularis, and alveolar septa. A small discrete mass in the right atrium consisted of similar neoplastic cells, and the spleen was diffusely infiltrated. Tissue distribution was suggestive of intravascular lymphoma. Neoplastic cells in tissue sections were immunoreactive for vimentin, CD18, CD45, and granzyme B and lacked immunoreactivity for cytokeratin. Neoplastic cells on bone marrow aspirate smears and blood films lacked immunoreactivity for CD3, CD79a, CD1c, CD11b, CD11c, CD11d, and E-cadherin. In the absence of immunophenotypic evidence for the neoplastic cells being derived from B-cell, T-cell, or histocytic/dendritic lineages and the lack of clonal antigen receptor gene rearrangement(s), along with positive immunoreactivity for granzyme B, a tumor of NK cells was considered likely. Based on current knowledge, this is the first report of canine intravascular lymphoma, of probable NK cell origin, with peripheral blood involvement.     

Uroplakin expression in the urothelial tumors of cows.

Expression of uroplakins (UPs) was investigated in 20 bladder tumors from cows that had been suffering from chronic enzootic hematuria for several years. In dysplastic urothelium and papillomatous proliferations, UP expression was evident both as luminal and intercellular staining. UPs appeared to clearly define the plasma membrane of luminal cells and the borders of cells placed in deeper layers, whether or not these intermediate cells were adjacent to superficial ones. Occasionally, some intermediate cells showed a remarkable cytoplasmic immunoreactivity. The pattern of UPs in grade I tumors was characterized by an evident discontinuity of luminal staining and by the presence of numerous intermediate cells showing a diffuse intracytoplasmic positivity for UPs. In grade II tumors, there was a decrease of luminal and intermediate cells showing UP expression and an apparent increase of clusters of intermediate cells with intracytoplasmic reactivity for UPs. In grade III tumors, immunoreactivity was heterogeneously distributed and a severe loss of UP-positive luminal and intermediate cells could be seen. Focally, superficial and deeper cells showed strong membraneous immunoreactivity that marked and delimited single cells, with complete circumferential peripheral staining clearly evident. UP expression in bladder tumors of cows reported in this study is similar to the UP pattern of some urothelial tumors in humans. Although UP expression is remarkably changed in bladder carcinogenesis of cattle, the UP gene(s) remains expressed during cell transformation and tumor progression.     

用单抗研究T细胞与牛流行性白血病发生的关系

本试验对10头患流行性白血病病牛的不同器官的肿瘤组织，用8种单克隆抗体（McAb),TH14B、BAQ44A、PIg45A、BIg715A、PIg501E、cAct105、MM1A、AHCC125染色进行了观察。结果：病牛经临床病理学和免疫琼扩试验而诊断为EBL，通过免疫组织化学检测，在10个病例中发现有9个病例的肿瘤组织对B细胞属性的McAb有很强的染色反应，证明其来源于B细胞。仅有1个病例对B细胞属性的McAb着色很淡，而对T细胞属性的McAb着色很深，呈强阳性反应，说明其来源可能与T细胞有关。     

Melanotic neurofibroma in a steer

A melanotic neurofibroma in a steer was investigated histologically, immunohistochemically and ultrastructurally. A very large tumor mass was located in the region of the head and right cheek. The tumor tissue consisted of an admixture of cells resembling Schwann cells and spindle-shaped cells, and they frequently contained melanin granules. Neoplastic Schwann cells were positive for S100 protein, with variation in intensity of staining, but most spindled cells were S100 negative. The tumor cells displayed ultrastructural features similar to those of Schwann cells or perineurial cells. The presence of melanosomes in varying stages of melanization in both cell types suggests that they have a common origin. This is a tumor of neural crest origin showing schwannian and perineurial differentiation, with ectopic production of melanin granules.     

Morphologic and immunohistochemical features of two spontaneous peripheral nerve tumors in Wistar rats

Morphologic features and S-100 protein immunoreactivity of a benign and malignant peripheral nerve sheath tumor were studied in two Wistar rats. Neoplasms that developed in untreated control rats from tumor bioassays were S-100 protein positive and had similar histopathologic features. Each peripheral nerve sheath tumor was encapsulated and composed of spindle cells arranged around small thin-walled blood vessels. Palisaded tumor cells were in the benign peripheral nerve sheath tumor while cells of the malignant peripheral nerve sheath tumor had cellular atypia and moderate numbers of mitoses. Ultrastructural examination of the malignant peripheral nerve sheath tumor revealed cells with external lamina and interdigitation of cytoplasmic processes. Intracytoplasmic concentric lamellae were seen; they were regularly spaced with a periodicity of about 15 nm. Such structures, indistinguishable from myelin sheaths, have not been commonly associated with peripheral nerve sheath tumors in man. Electron microscopy and immunohistochemistry were useful in the diagnosis of these tumors as Schwannomas and in differentiation from other spindle cell tumors.     

A collision tumor consisting of granular cell tumor and adenocarcinoma in the uterus of an aged djungarian hamster

A neoplastic nodular lesion consisting of an admixture of granular cell tumor and adenocarcinoma was found in the uterus of a 26-month-old Djungarian hamster. Neoplastic cells of the uterine adenocarcinoma showed an epithelial nature in their growth patterns and by cytokeratin-immunopositive reaction, exhibiting nuclear pleomorphism. The granular cells had an abundant amount of fine granular eosinophilic cytoplasm and eccentric or central nuclei with no nuclear atypia; the granular structures were positive for periodic acid-Schiff with diastase resistance and were confirmed as lysosomes/autophagosomes by electron microscopy; immunohistochemically, the cells reacted to desmin, vimentin and α-smooth muscle actin and negatively for neurogenic, histiocyte/macrophage or epithelial markers, indicating smooth muscle origin. Because these tumors were generated from different cell origins, a diagnosis of collision tumor was made.     

MicroRNA-21 expression,serum tumor markers,and immunohistochemistry in canine mammary tumors

Background

Canine mammary tumors (CMTs) are one of the most common malignancies in dogs and are associated with significant mortality. Serum tumor markers and non-coding microRNAs have gained widespread popularity in human oncology studies. The present study has two aims, first one is to investigate the miR-21 expression compared with changes in serum tumor markers (CEA and CA15-3) in CMT. The second aim is to detect the immunohistochemistry markers as vimentin, P63, and -SMA in CMT.

Methods

This study enrolled 17 female dogs: 10 with mammary tumors and seven controls without tumors. Blood samples were collected to measure miR-21, CEA, and CA 15-3, and histological samples were prepared for histological grading and immunohistochemistry.

Results

CA 15-3 was elevated in all animals, whereas CEA levels showed no change compared with controls. miR-21 was upregulated 12.84-fold in animals with CMT. The most frequently recorded CMT was the mixed type. Myoepithelial cells were identified by P63 immunoreactivity, but not SMA. High expression of miR-21 was observed with positive vimentin immunoreactivity, indicating the mesenchymal origin of the tumor cells.

Conclusion

The present study showed that miR-21 was elevated to a greater extent than CA 15-3 (12.84-fold vs. threefold). Tumors that was positive for vimentin immunoreactivity was also associated with an elevation in the levels of miR-21, showing that miR-21 is released from mesenchymal cells. These findings support the hypothesis that miR-21 may be a more sensitive, noninvasive indicator for CMT.

     

Establishment and characterization of a cell line, MCO-Y4, derived from canine mammary gland osteosarcoma

A cell line, MCO-Y4, was established from a mammary gland osteosarcoma of a 16-year-old female mongrel dog. Histopathologically the tumor was composed of osteoblastic cells with an osteoid meshwork and chondroid matrix. The mean doubling time of the cells at the 93rd passage was 32.39+/-4.66 hr. Immunohistochemically, the osteoblastic and chondroblastic cells were positive for bone morphogenetic protein (BMP)-2/4 and BMP receptor (BMPR) II. The cultured cells were spindle in shape during the growth and the confluent phases. No tumor matrix was detected in the culture dish by alcian blue staining or von-Kossa silver impregnation. MCO-Y4 cells on the chamber slides showed intense immunoreactivity for BMP-2/4 and BMPR II. Noggin, an antagonist for BMP-2/4, showed the growth inhibition on MCO-Y4 cells. In addition, fibronectin might be potential for stimulating growth of MCO-Y4 cells. When transplanted into severe combined immunodeficiency mice, the cells formed tumors consisting of solid proliferation of osteoblastic and fibroblastic cells with woven-bone trabeculae. These tumor cells were intensely positive for BMP-2/4 and BMPR II. Our results suggested that the cell line might be useful for studying the role of BMPs in canine osteosarcoma and the mechanism of ossification.     

T-cell lymphoma with eosinophilic infiltration involving the intestinal tract in 11 dogs

Among the intestinal tumors of hematopoietic cell origin, lymphoma is the most common in the dog. Herein, we characterized the clinical and pathologic features of 11 dogs (average age, 10.6 +/- 2.5 years) with T-cell lymphoma of the intestinal tract with eosinophil infiltrates. No sex predominance was apparent. All had localized tumor masses in the small intestine. Grossly, the intestinal wall was thickened, and the lumen of the affected intestine was usually narrowed. Microscopically, we observed transmural diffuse invasion of round to pleomorphic tumor cells. Tumor cells showed varying morphology, from scanty to abundant cytoplasm, and round to ovoid nuclei with scattered to dense chromatin. In seven of the dogs, tumor cells had infiltrated into the epithelium. All showed infiltration of eosinophils and all 11 tumors had a T-cell phenotype (CD3+, CD79-). Only one tumor stained positive for the mast cell marker c-kit and none was positive for mast cell tryptase. We did not observe ultrastructurally apparent granules in any of the tumor cells. These results suggest that, in dogs, T-cell lymphomas of intestinal origin resemble mast cell tumors of intestinal origin with respect to cell structure and eosinophil infiltration. Therefore, in the absence of epitheliotropism, it is difficult to confirm the differential diagnosis without immunostaining for mast cell and lymphocyte markers, including mast cell tryptase, c-kit, CD3, and CD79.     

Malignant histiocytosis in cattle

Malignant histiocytosis was diagnosed in 4 cows. In all cases the tumor tissues were composed of cytologically atypical histiocytes with evidence of erythrophagocytosis. The tumor in case 1 appeared highly anaplastic with marked nuclear pleomorphism, and had areas of spindle cell differentiation, but had no relation to malignant fibrous histiocytoma. The neoplastic tissue in case 2, characterized by cohesive growth of tumor cells, was distinguishable from anaplastic carcinoma cells by cytokeratin immunostaining. There were many hemosiderin-laden neoplastic cells suggestive of high phagocytic activity in a lymph node of case 3. The neoplastic cells in case 4, frequently multinucleated, were less atypical than in the other cases. All cases expressed histiocyte-associated markers (lysozyme and HAM56), and were negative for cytokeratin, S100, and T- and B-cell lineage-specific markers (CD3 and CD79a). The most frequent HAM56 immunoreactivity was detected in case 4, and the giant, multinucleated forms, reminiscent of epithelioid cell differentiation. seemed not to indicate cytological pleomorphism as a result of neoplastic transformation.     

Giant cell tumour of bone in a cat with extraskeletal metastases: pathological and immunohistochemical study

A case of giant cell tumour of bone (GCTb) in the lung and in a subcutaneous mass located in the right flank, with a probable primary origin in the mid-diaphysis of the right tibia, was described in a 8-year-old female cat. Numerous multinucleated giant cells were homogeneously distributed among a population of ovoid or spindle-shaped mononuclear cells. All of them were positive for vimentin suggesting a mesenchymal origin. Spindle-shaped tumour cells resemble fibroblastic cells, showing collagen fibres in their vicinity. Ovoid mononuclear cells are similar to macrophages, with a cytoplasm rich in electron-dense lysosomes. Multinucleated giant cells appear morphologically similar to osteoclasts. These findings are supported for the positive reaction to tartrate-resistant acid phosphatase (TRAP) and lysozyme, encountered only in ovoid and multinucleated giant cells. No immunoreactivity against human oestrogen receptors was observed in the nuclei of any neoplastic cells.     

Osteoclast-like giant cell tumour arising from the kidney in a dog

In this study, a case of osteoclast-like giant cell tumour arising from the kidney is reported in an eight-year-old female Anatolian Shepherd dog. Macroscopically, the tumorous mass covered the hilus of the left kidney. It was 26 x 22 x 12 cm in size and 3700 g in weight. Metastatic tumorous nodules, 0.5-2.0 cm in diameter, were found on the abdominal side of the diaphragm and in the lungs. Microscopically, numerous large osteoclast-like multinucleated giant cells and spindle-spheroidal-shaped cells were seen. Osteoblastic differentiation and osteoid matrix were noted in a few areas at the periphery of the tumour, near the connective tissue septa. The stroma of the tumour tissue was vascular, oedematous and loose. By immunoperoxidase staining, tumour cells showed immunoreactivity for vimentin but not for keratin and desmin, indicating that the tumour had mesenchymal origin. This is the first report in the literature on a malignant osteoclast-like giant cell tumour arising from a visceral organ in animals.     

Immunohistochemical profiling and telomerase activity of a canine medulloblastoma

A well-demarcated mass was found by computed tomography in the left cerebellar hemisphere of a 4-year-old male Boxer with acute onset of progressive central vestibular syndrome. At necropsy, the pink, gelatinous mass was in the flocculonodular lobe. Histologically, neoplastic tissue arose from the granular layer of the cerebellar cortex and consisted of sheets of oval to round hyperchromatic cells, consistent with the diagnosis of medulloblastoma. Synaptophysin and neuron-specific enolase immunoreactivity supported the neuronal origin of the neoplastic cells; furthermore, a weak to moderate c-kit expression was detected, as reported in pediatric medulloblastoma. Telomerase activity of tumor cells was demonstrated by immunohistochemistry and by the telomere repeat amplification protocol, suggesting involvement of this enzymatic pathway.     

Immunohistochemical localization of insulin-like growth factor-I receptor (IGF-IR) in the developing and mature rat testes

It has been suggested that insulin-like growth factor-I (IGF-I) plays an important role in the regulation of spermatogenesis in the testes. Its signal is mediated predominantly by the IGF-I receptor (IGF-IR). Signalling through IGF-IR has been shown to have a potent survival function. IGF-IR, a transmembrane tyrosine kinase, is widely expressed across many cell types. In this study, we demonstrated the distribution of IGF-IR in testes of differently aged rats. Anti-IGF-IR is a rabbit polyclonal antibody raised against a peptide mapping at the carboxy terminus of the IGF-IR of human origin. Testicular specimens were fixed in Bouin's solution and embedded in paraffin. The paraffin-embedded sections were processed for standard immunohistochemistry by the labelled streptavidin-biotin technique. At postnatal day 19, IGF-IR immunoreactivity was seen moderately in spermatogonia, and slightly both in leptotene and zygotene primary spermatocytes. At postnatal day 35, immunoreactivity was seen slightly both in the pachytene primary spermatocytes and Leydig cells. Although there was intense immunoreactivity in the Leydig cells and in the elongated spermatids on days 50 and 70, the intensity of reaction was decreased in the elongated spermatids in the 10th month. Our results suggest that IGF-IR may play significant roles in testicular function and germ cell development.     

Cytologic variants of γδ T cell lymphoma in cattle

Two cytologic variants of γδ T cell lymphoma are described. Case 1 represented a giant cell variant found in a 5-year-old Holstein cow, which had large tumor masses in the pelvic cavity. This variant consisted of very large lymphoid cells with round to oval nuclei, medium-sized nucleoli and abundant cytoplasm. Case 2 was an aborted 7-month-old female Holstein fetus, which represented an immature cell variant. Most of the neoplastic lesions were located in the skin and pleural and peritoneal submesothelial tissues. The neoplastic tissues were composed of homogeneous growth of lymphoma cells characterized by inconspicuous nucleoli and finely dispersed chromatin. Both cases demonstrated CD3, CD8 and WC1 immunoreactivity. The current study revealed that there are 4 cytologic variants (common, giant cell, hypergranular and immature cell) in bovine γδ T cell lymphomas.