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Fig. 1a. Skin; canine, mast cell tumor. Immunohistochemical staining for tryptase antigen detection demonstrating numerous positive tumor cells. BMK iVIEW DAB Paraffin detection kit (Ventana Medical Systems Inc.) with primary antibody AA1 (1 : 50, DAKO). Bar 50 m. Fig. 1b. Skin; canine, mast cell tumor. Histochemical staining for chymase activity demonstrating numerous positive tumor cells. AS-D chloroacetate substrate, Procedure No. 91 (Sigma-Aldrich Canada Ltd.). Bar 50 m. Fig. 2. Skin; canine, mast cell tumor. Immunohistochemical staining for CD18 detection demonstrating numerous positive tumor cells (arrows). Note that staining is membranous rather than cytoplasmic. Cells demonstrating cytoplasmic staining are eosinophils (arrowheads). This tumor was also positive with toluidine blue, but negative for tryptase antigen and chymase activity. BMK iVIEW DAB Paraffin detection kit (Ventana Medical Systems Inc.) with primary antibody CA16.3C10 (1 : 4, P. Moore). Bar 50 m. Fig. 3. Fig. 3a. Skin; canine, histiocytoma. Immunohistochemical staining for CD18 detection. BMK iVIEW DAB Paraffin detection kit (Ventana Medical Systems Inc.) with primary antibody CA16.3C10 (1 : 4, P. Moore). Bar 50m. Fig. 3b. Skin; canine, histiocytoma. Immunohistochemical staining for MHC class II detection. BMK iVIEW DAB Paraffin detection kit (Ventana Medical Systems Inc.) with primary antibody TAL.1B5 (1 : 150, DAKO). Bar 50 m. Fig. 4. Skin; canine, regressing histiocytoma or T-cell lymphosarcoma. Immunohistochemical staining for CD3 detection. Note that positive cells do not have typical lymphocyte morphology (arrows). BMK iVIEW DAB Paraffin detection kit (Ventana Medical Systems Inc.) with primary antibody CD3-12 (1 : 25, Novocastra Laboratories Ltd.). Bar 50 m.
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Immunohistochemical and histochemical stains are useful adjunct techniques in the diagnosis of canine cutaneous round cell tumors, which can appear histologically similar. We applied a panel of monoclonal antibodies (recognizing tryptase, chymase, serotonin for mast cells; CD1a, CD18, MHC class II for histiocytes; CD3 for T lymphocytes; CD79a for B...
Citations
... Sections were then counterstained with Mayer's hematoxylin and rinsed in distilled water (Fernandez et al., 2005). Leica DM500 microscope and a digital camera (Leica EC3, Leica, Germany) were used to take micrograph slices. ...
Round cell tumors are common cutaneous lesions in dogs, with increased occurrence percentages among different skin tumors. This study aimed to investigate the frequency as well as gross and pathological characteristics of round cell tumors in natural cases of tumorous dogs in relation to breed, sex, and age. Moreover, it aimed to evaluate the immunohistochemical expression of a panel of immunohistochemical stains, including vimentin, E-cadherin, and cluster of differentiation (CD45) as an adjunct technique for the differential diagnosis of cutaneous round cell neoplasm. Data were collected from 64 dogs of both sexes (36 females and 28 males), various breeds, and different ages (8 months to 7 years). The histopathological nature of neoplastic growth was reported, and neoplasm prevalence was classified using age, sex, breed, and site on the body. We observed 48 cases of transmissible venereal tumors, 12 cutaneous histiocytomas, and 4 histiocytic sarcoma. Immunohistochemical characterization revealed an intense positive immunoreactivity for vimentin in transmissible venereal tumor cells and moderate positive immunoreactivity for E-cadherin and CD45 in cutaneous histiocytoma and histiocytic sarcoma cells. In conclusion, the canine transmissible venereal tumor was the most frequent form of round cell tumor; thus, a definitive cutaneous neoplasm diagnosis should be based on histopathological morphology and immunohistochemical findings.
... The disease starts as localized lesions that spread quickly to numerous organs, including the spleen, lungs, skin, brain (meninges), lymph nodes, and bone marrow in addition to the synovial tissues of the limbs (Moore, 2014). Based only on histology, it can be challenging to distinguish CCH from plasmacytomas, mast cell tumors, and cutaneous lymphomas due to its morphologic similarity to other round-cell tumors (Fernandez et al., 2005;Araújo et al., 2012). According to previous research (Schwens et al., 2011;Moore, 2014), the majority of the assessed CCHs were discovered in dogs under the age of five and were situated in the head, neck, limbs, or trunk. ...
Background
Cutaneous neoplastic disorders are often observed in small mammal pets, such as dogs, regardless of their gender.
Aim
An important objective of this work was to give a full account of the clinical, pathological, and immune-histochemical features of several skin tumors in dogs.
Methods
This study was a case series in the hospital clinic, Faculty of Veterinary Medicine, Zagazig University, Egypt. Twenty-five dogs (14 males and 11 females) were examined clinically during the period from March 2022 to October 2023. The skin swelling was collected from affected animals and then subjected to a detailed histopathological study to record the different gross and microscopic findings and confirm the diagnosis by immunohistochemistry.
Results
Skin neoplasia in dogs was exposed to various clinical signs, and the dogs' ages ranged between 3 and 11 years. Concerning tumor features, the majority of neoplasms were malignant (65.52%) more than benign (34.48%). The study revealed the presence of 29 cases of dogs showed neoplasia with different prevalence rates including squamous cell carcinoma (13.79%), mast cell tumor (6.89%), basal cell tumors (10.34%), histiocytoma (6.89%), trichoepithelioma (10.34%), transmissible venereal tumor (10.34%), trichilemmoma (3.44%), scalp paraganglioma (3.44%), pilomatricoma (10.34%), malignant melanomas (17.24%), and miscellaneous cases as fat necrosis (6.89%), in males and females dogs with different histopathological lesions and immunohistochemistry expressions for pan-cytokeratin (CK), melanocyte-differentiation antigens (S100 protein), and synaptophysin.
Conclusion
Malignant melanomas (17.24%) are the extremely common cutaneous tumors diagnosed in this study. Meanwhile, benign tumors such as trichilemmoma, trichoepithelioma, pilomatricoma, and paraganglioma are less frequent in dogs.
... The disease starts as localized lesions that spread quickly to numerous organs, including the spleen, lungs, skin, brain (meninges), lymph nodes, and bone marrow in addition to the synovial tissues of the limbs (Moore, 2014). Based only on histology, it can be challenging to distinguish CCH from plasmacytomas, mast cell tumors, and cutaneous lymphomas due to its morphologic similarity to other round-cell tumors (Fernandez et al., 2005;Araújo et al., 2012). According to previous research (Schwens et al., 2011;Moore, 2014), the majority of the assessed CCHs were discovered in dogs under the age of five and were situated in the head, neck, limbs, or trunk. ...
Background: Cutaneous neoplastic disorders are often observed in small mammal pets, such as dogs, regardless of
their gender.
Aim: An important objective of this work was to give a full account of the clinical, pathological, and immunehistochemical features of several skin tumors in dogs.
Methods: This study was a case series in the hospital clinic, Faculty of Veterinary Medicine, Zagazig University,
Egypt. Twenty-five dogs (14 males and 11 females) were examined clinically during the period from March 2022 to
October 2023. The skin swelling was collected from affected animals and then subjected to a detailed histopathological
study to record the different gross and microscopic findings and confirm the diagnosis by immunohistochemistry.
Results: Skin neoplasia in dogs was exposed to various clinical signs, and the dogs' ages ranged between 3 and 11
years. Concerning tumor features, the majority of neoplasms were malignant (65.52%) more than benign (34.48%).
The study revealed the presence of 29 cases of dogs showed neoplasia with different prevalence rates including
squamous cell carcinoma (13.79%), mast cell tumor (6.89%), basal cell tumors (10.34%), histiocytoma (6.89%),
trichoepithelioma (10.34%), transmissible venereal tumor (10.34%), trichilemmoma (3.44%), scalp paraganglioma
(3.44%), pilomatricoma (10.34%), malignant melanomas (17.24%), and miscellaneous cases as fat necrosis (6.89%),
in males and females dogs with different histopathological lesions and immunohistochemistry expressions for pancytokeratin (CK), melanocyte-differentiation antigens (S100 protein), and synaptophysin.
Conclusion: Malignant melanomas (17.24%) are the extremely common cutaneous tumors diagnosed in this study.
Meanwhile, benign tumors such as trichilemmoma, trichoepithelioma, pilomatricoma, and paraganglioma are less
frequent in dogs.
Keywords: Dogs, Skin tumors, Trichoepithelioma, Paraganglioma, Melanoma
... Características como grande volume citoplasmático, cor clara, núcleos irregulares e ausência de corpos irregulares, são essenciais na diferenciação de células linfóides e células de histiocitoma (MEINKOTH et al, 2014). Em caso da não utilização da imuno-histoquímica, histologicamente é possível notar que o histiocitoma possui potencial para ISSN: 2675-8008 V. 4, Nº 3, 2023 DOI: 10.51161/clinvet2023/19805 invadir a epiderme e se diferenciar das outras neoplasias de células redondas (FERNANDEZ et al., 2005), fato observado no presente estudo. O prognóstico do histiocitoma cutâneo canino é favorável devido à sua regressão espontânea que ocorre normalmente em até 3 meses ainda de maneira não esclarecida, porém entende-se que possa estar relacionada com a migração de células para os linfonodos que são responsáveis pela drenagem da pele. ...
... Haematoxylin and Eosin, Goldner's Trichrome stains. Tissue sections were further labelled immunohistochemically for B lymphocytic cells using anti-CD20 antibody (Degl'Innocenti et al., 2019) and anti-CD79aantibody(Fernandez et al., 2005) and T lymphocytic cells by an anti-CD3 antibody(Fernandez et al., 2005) and histiocytic cells by anti-Iba1 antibody(Ohara et al., 2019), following a routine protocol. Briefly, 4-µm-thick sections were de-waxed in xylene, hydrated throughout a graded series of ethanol and rehydrated in deionised water. ...
... Haematoxylin and Eosin, Goldner's Trichrome stains. Tissue sections were further labelled immunohistochemically for B lymphocytic cells using anti-CD20 antibody (Degl'Innocenti et al., 2019) and anti-CD79aantibody(Fernandez et al., 2005) and T lymphocytic cells by an anti-CD3 antibody(Fernandez et al., 2005) and histiocytic cells by anti-Iba1 antibody(Ohara et al., 2019), following a routine protocol. Briefly, 4-µm-thick sections were de-waxed in xylene, hydrated throughout a graded series of ethanol and rehydrated in deionised water. ...
A 5‐year‐old spayed female American Staffordshire was referred for weakness, reluctance to move and distension of the abdomen. Three weeks before, the dog underwent surgery for excision of a nodular mass suspected to be a non‐epitheliotropic cutaneous T‐cell lymphoma (NE‐CTCL). Computed tomography revealed heterogeneous enhancing mesenteric masses and nodular lesions of soft tissue density, and infiltration of the abdominal muscular wall. Moreover, a pattern of diffuse muscle nodules in the skeletal muscles was visible, with lesions showing homogenous, heterogeneous or ring enhancement. Necrosis was histologically observed and these lesions were infiltrated by CD3‐positive and CD20‐, CD79a‐ and Iba1‐negative neoplastic lymphocytes. On the basis of the immunopathological features metastatic NE‐CTCL was suspected. Skeletal muscle metastasis has been rarely reported in small animals and this case report further confirms that this possibility should be considered in dogs with lymphoma.
... Para fazer o diagnóstico dessas neoplasias o exame histopatológico e a técnica de imunohistoquímica se tornam a melhor ferramenta para elucidação do prognóstico e conseguintemente melhor eficácia no tratamento da doença (Fernandez et al., 2005;Silva et al., 2015). É importante lembrar que na medicina veterinária a técnica de imuno-histoquímica ainda não é utilizada em todos os casos devido ao seu custo e à escassez de anticorpos espécie-específicos em algumas situações. ...
... De acordo com Fernandez et al. (2005) a diferenciação não seja conclusiva. E neste caso, é possível perceber que com elevado grau de diferenciação o histopatológico como único exame não é suficiente para concluir com exatidão os diferentes tipos de tumores de células redondas, dando como diagnóstico a possibilidade de plasmocitoma e sugerindo a realização da imuno-histoquímica. ...
Os tumores de células redondas consistem na semelhança morfológica entre as células que compõem esses tumores, que são classificados quanto a origem embriológica em neoplasias mesenquimais. Entre os diferentes tipos neoplásicos provenientes das células redondas este trabalho dará ênfase para o mieloma múltiplo e o linfoma. O mieloma múltiplo é uma neoplasia rara, que tem origem da proliferação neoplásica de plasmócitos a partir da medula óssea. O linfoma é uma neoplasia com origem nos tecidos hematopoiéticos sólidos e a etiologia em cães ainda é desconhecida. Desta forma, o objetivo deste trabalho foi relatar um animal com diagnostico de linfoma cutâneo não epteliotrópico de imunofenótipo T apresentando aspectos clínicos evidentes de mieloma múltiplo.
... Especially round cell tumors, which can have similar morphologies, are oftentimes hard to distinguish 1, 2 . Tumorspecific immunohistochemical (IHC) stainings can help to distinguish tumor subtypes but are considerably more expensive and time-consuming and still might not provide reliable results for undifferentiated tumors 1 . Deep learning-based algorithms can support the pathologist in segmenting and classifying cutaneous tumors using standard Hematoxylin & Eosin (H&E) stain and have successfully been applied in various works [3][4][5][6][7][8][9] . ...
Due to morphological similarities, the differentiation of histologic sections of cutaneous tumors into individual subtypes can be challenging. Recently, deep learning-based approaches have proven their potential for supporting pathologists in this regard. However, many of these supervised algorithms require a large amount of annotated data for robust development. We present a publicly available dataset consisting of 350 whole slide images of seven different canine cutaneous tumors complemented by 12,424 polygon annotations for 13 histologic classes including seven cutaneous tumor subtypes. Regarding sample size and annotation extent, this exceeds most publicly available datasets which are oftentimes limited to the tumor area or merely provide patch-level annotations. We validated our model for tissue segmentation, achieving a class-averaged Jaccard coefficient of 0.7047, and 0.9044 for tumor in particular. For tumor subtype classification, we achieve a slide-level accuracy of 0.9857. Since canine cutaneous tumors possess various histologic homologies to human tumors, we believe that the added value of this dataset is not limited to veterinary pathology but extends to more general fields of application.
... Chymase and tryptase can be used as additional immunohistochemical markers both in differentiating canine mast cell subpopulations in tissues 20,46 and in identifying MCTs. 8 In addition to tryptase and chymase, mast cells in humans, mice, and rats express an additional mast cell-specific secretory protease, CPA3, 33 also called MC-CPA. CPA3 expression is considered to be restricted to mast cell secretory granules, with the exception of reported CPA3 expression on RNA level in basophils of patients with allergic conditions. ...
Mast cell tumors (MCTs) are one of the most common cutaneous malignancies in dogs. Previous studies have reported expression of mast cell–specific proteases chymase and tryptase in canine cutaneous MCTs and in connective tissue and mucosal mast cells. In humans and rodents, mast cells express an additional specific protease, carboxypeptidase A3 (CPA3). In this article, we describe CPA3 immunoreactivity in connective tissue, visceral, mucosal, and neoplastic mast cells in dogs. Positive immunolabeling for CPA3 was observed in nonneoplastic mast cells in 20/20 formalin-fixed paraffin-embedded normal tissues (skin, liver, spleen, intestine), and in 63/63 MCTs irrespective of their histological grade. CPA3 protein expression was comparable to that of c-kit in both the nonneoplastic and neoplastic mast cells. Three distinct labeling patterns (membranous, diffuse, and focal cytoplasmic) were observed for CPA3 in MCTs. The focal cytoplasmic labeling pattern was associated with high-grade MCTs staged with the Kiupel 2-tier grading criteria. We propose CPA3 as a novel immunohistochemical marker for canine mast cells in health and disease.
... In rodents, serotonin was found, primarily, in connective tissue mastocytes [33,141]. Some researchers failed to find serotonin using immunohistochemistry in mast cells of cat and dog skin [138,142], and in the brain of experimental animals and birds it was revealed only in some mast cells [120,143,144]. On the other hand, serotonin can be found (apart from mastocytes) in many other cell types of human and animal organs; among them are enterochromaffin cells of the gastrointestinal tract, which produce the great majority of serotonin in the body; various endocrine cells of the lungs, adrenals, prostate, epididymis [145][146][147][148]; serotonergic neurons of the central and peripheral nervous system [149,150]; type III gustatory receptor cells and glomus cells of carotid body [151,152]. ...
Mast cells play an important role in the body defense against allergens, pathogens, and parasites by participating in inflammation development. However, there is evidence for their contributing to the pathogenesis of a number of atopic, autoimmune, as well as cardiovascular, oncologic, neurologic, and other diseases (allergy, asthma, eczema, rhinitis, anaphylaxis, mastocytosis, multiple sclerosis, rheumatoid arthritis, inflammatory gastrointestinal and pulmonary diseases, migraine, etc.). The diagnosis of many diseases and the study of mast cell functions in health and disease require their identification; so, the knowledge on adequate imaging techniques for mast cells in humans and different species of animals is of particular importance.
The present review summarizes the data on major methods of mast cell imaging: enzyme histochemistry, immunohistochemistry, as well as histochemistry using histological stains. The main histological stains bind to heparin and other acidic mucopolysaccharides contained in mast cells and stain them metachromatically. Among these are toluidine blue, methylene blue (including that contained in May-Grünwald–Giemsa stain), thionin, pinacyanol, and others. Safranin and fluorescent dyes: berberine and avidin — also bind to heparin. Longer staining with histological dyes or alcian blue staining is needed to label mucosal and immature mast cells.
Advanced techniques — enzyme histochemistry and especially immunohistochemistry — enable to detect mast cells high-selectively using a reaction to tryptases and chymases (specific proteases of these cells). In the immunohistochemical study of tryptases and chymases, species-specific differences in the distribution of the proteases in mast cells of humans and animals should be taken into account for their adequate detection. The immunohistochemical reaction to immunoglobulin E receptor (FcεRI) and c-kit receptor is not specific to mast cells, although the latter is important to demonstrate their proliferation in normal and malignant growth.
Correct fixation of biological material is also discussed in the review as it is of great significance for histochemical and immunohistochemical mast cell detection.
Fluorescent methods of immunohistochemistry and a multimarker analysis in combination with confocal microscopy are reported to be new technological approaches currently used to study various mast cell populations.
... While our findings demonstrate a role for CD18 in platelet and RBC extravasation, a variety of leukocytes express CD18, including mast cells [26][27][28]. Electron microscopy of corneas following epithelial abrasion in WT mice has shown that PMNs, platelets, and RBCs extravasated from the limbal venules and PMNs came into contact with perivascular mast cells ( Figure 3A). Since mast cells also express CD18, we performed additional experiments to determine whether mast cells mediated platelet and RBC extravasation. ...
Platelet extravasation during inflammation is under-appreciated. In wild-type (WT) mice, a central corneal epithelial abrasion initiates neutrophil (PMN) and platelet extravasation from peripheral limbal venules. The same injury in mice expressing low levels of the β2-integrin, CD18 (CD18hypo mice) shows reduced platelet extravasation with PMN extravasation apparently unaffected. To better define the role of CD18 on platelet extravasation, we focused on two relevant cell types expressing CD18: PMNs and mast cells. Following corneal abrasion in WT mice, we observed not only extravasated PMNs and platelets but also extravasated erythrocytes (RBCs). Ultrastructural observations of engorged limbal venules showed platelets and RBCs passing through endothelial pores. In contrast, injured CD18hypo mice showed significantly less venule engorgement and markedly reduced platelet and RBC extravasation; mast cell degranulation was also reduced compared to WT mice. Corneal abrasion in mast cell-deficient (KitW-sh/W-sh) mice showed less venule engorgement, delayed PMN extravasation, reduced platelet and RBC extravasation and delayed wound healing compared to WT mice. Finally, antibody-induced depletion of circulating PMNs prior to corneal abrasion reduced mast cell degranulation, venule engorgement, and extravasation of PMNs, platelets, and RBCs. In summary, in the injured cornea, platelet and RBC extravasation depends on CD18, PMNs, and mast cell degranulation.
