Immunohistochemical and Histochemical Stains for Differentiating Canine Cutaneous Round Cell Tumors

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 lymphocytes and plasma cells) and one histochemical stain (naphthol AS-D chloroacetate for chymase activity) to formalin-fixed, paraffin-embedded sections of canine cutaneous mast cell tumors, histiocytomas, lymphosarcomas, plasmacytomas, and unidentified round cell tumors. Of 21 tumors with a histologic diagnosis of mast cell tumor, 7/7 (100%) grade I, 6/7 (85.7%) grade II, and 3/7 (42.9%) grade III tumors were diagnosed as mast cell tumors based on positive staining for tryptase antigen and chymase activity. Mast cells were positive for both tryptase antigen and chymase activity, indicating equal efficacy of tryptase immunohistochemistry and chymase histochemistry. Chymase was detected immunohistochemically in both tumor and nontumor cells, while serotonin was not detected in most mast cell tumors, and thus, neither was useful in the diagnosis of mast cell tumors. Immunohistochemistry to detect CD18 and MHC class II was equally effective in staining histiocytomas, although lymphosarcoma must be ruled out through the use of CD3 and CD79a immunohistochemistry. Immunohistochemistry using three different monoclonal antibodies to human CD1a showed no cross-reactivity in canine histiocytomas and was not useful. A final diagnosis was obtained for 4/5 (80%) of the unidentified tumors, indicating the usefulness of multiple stains in poorly differentiated round cell tumors.

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Cytology of Canine Cutaneous Round Cell Tumors

Veterinary Pathology ◽

10.1177/030098587901600605 ◽

1979 ◽

Vol 16 (6) ◽

pp. 673-679 ◽

Cited By ~ 35

Author(s):

J. R. Duncan ◽

K. W. Prasse

Keyword(s):

Mast Cell ◽

Final Diagnosis ◽

Cell Tumor ◽

Histologic Examination ◽

Round Cell ◽

Mast Cell Tumors ◽

Mast Cell Tumor ◽

Round Cell Tumors

Sixty-four canine cutaneous round cell tumors were divided into 25 mast cell tumors, 15 histiocytomas, nine cutaneous lymphosarcomas and 15 transmissible venereal tumors. The final diagnosis was made from cytologic, clinical and histologic findings. Cytologic features were significantly distinctive in mast cell tumor, transmissible venereal tumor, and most cases of histiocytoma and lymphosarcoma to allow a diagnostic opinion. This opinion was supported by subsequent histologic examination. In some instances cytology was considered essential in rendering a diagnostic opinion even though histology was available.

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Suppressible and nonsuppressible autocrine mast cell tumors are distinguished by insertion of an endogenous retroviral element (IAP) into the interleukin 3 gene.

Journal of Experimental Medicine ◽

10.1084/jem.178.2.403 ◽

1993 ◽

Vol 178 (2) ◽

pp. 403-411 ◽

Cited By ~ 18

Author(s):

H H Hirsch ◽

A P Nair ◽

C Moroni

Keyword(s):

Mast Cell ◽

Mast Cells ◽

Gene Transcription ◽

Class Ii ◽

Class I ◽

Ras Oncogene ◽

Interleukin 3 ◽

Mast Cell Tumors ◽

Intracisternal A Particle

After v-H-ras expression, the interleukin 3 (IL-3)-dependent PB-3c mast cells progress in vivo to two different classes of IL-3 autocrine tumors. Class I tumors show a germline configuration of the IL-3 gene and represent more than 90% of tumors analyzed so far. Somatic cell fusion of class I tumor lines with the nontumorigenic parental PB-3c resulted in loss of oncogenic IL-3 expression by a posttranscriptional mechanism with concomitant tumor suppression. Class II tumors arise rarely and contain an insertion in one IL-3 allele. This alteration was linked to enhanced IL-3 gene transcription. For one tumor, the insertion was shown to be an endogenous retroviral element (intracisternal A-particle). Cell hybrids of class II tumors with PB-3c remained IL-3 independent, expressed IL-3, and formed tumors rapidly. These results suggest that the v-H-ras oncogene synergizes with a recessive and a dominant lesion in class I and II tumors, respectively, both of which lead to the autocrine production of IL-3.

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Secretory granules of mast cells accumulate mature and immature MHC class II molecules

Journal of Cell Science ◽

10.1242/jcs.114.2.323 ◽

2001 ◽

Vol 114 (2) ◽

pp. 323-334

Author(s):

H. Vincent-Schneider ◽

C. Thery ◽

D. Mazzeo ◽

D. Tenza ◽

G. Raposo ◽

...

Keyword(s):

Mast Cell ◽

Mast Cells ◽

Mhc Class Ii ◽

Cell Activation ◽

Secretory Granules ◽

Intracellular Localization ◽

Class Ii ◽

Mast Cell Activation ◽

Invariant Chain ◽

Mhc Class Ii Molecules

Bone marrow-derived mast cells as well as dendritic cells, macrophages and B lymphocytes express major histocompatibility complex (MHC) class II molecules. In mast cells, the majority of MHC class II molecules reside in intracellular cell type-specific compartments, secretory granules. To understand the molecular basis for the localisation of MHC class II molecules in secretory granules, MHC class II molecules were expressed, together with the invariant chain, in the mast cell line, RBL-2H3. Using electron and confocal microscopy, we observed that in RBL-2H3 cells, mature and immature class II molecules accumulate in secretory granules. Two particular features of class II transport accounted for this intracellular localization: first, a large fraction of newly synthesized MHC class II molecules remained associated with invariant chain fragments. This defect, resulting in a slower rate of MHC class II maturation, was ascribed to a low cathepsin S activity. Second, although a small fraction of class II dimers matured (i.e. became free of invariant chain), allowing their association with antigenic peptides, they were retained in secretory granules. As a consequence of this intracellular localization, cell surface expression of class II molecules was strongly increased by cell activation stimuli which induced the release of the contents of secretory granules. Our results suggest that antigen presentation, and thereby antigen specific T cell stimulation, are regulated in mast cells by stimuli which induce mast cell activation.

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Histopathological Classification of Canine Cutaneous Round Cell Tumors Using Deep Learning: A Multi-Center Study

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.640944 ◽

2021 ◽

Vol 8 ◽

Author(s):

Massimo Salvi ◽

Filippo Molinari ◽

Selina Iussich ◽

Luisa Vera Muscatello ◽

Luca Pazzini ◽

...

Keyword(s):

Mast Cell ◽

Deep Learning ◽

Learning Strategy ◽

Computational Time ◽

Round Cell ◽

Test Set ◽

Histopathological Classification ◽

Mast Cell Tumors ◽

Automated Algorithm ◽

Round Cell Tumors

Canine cutaneous round cell tumors (RCT) represent one of the routine diagnostic challenges for veterinary pathologists. Computer-aided approaches are developed to overcome these restrictions and to increase accuracy and consistency of diagnosis. These systems are also of high benefit reducing errors when a large number of cases are screened daily. In this study we describe ARCTA (Automated Round Cell Tumors Assessment), a fully automated algorithm for cutaneous RCT classification and mast cell tumors grading in canine histopathological images. ARCTA employs a deep learning strategy and was developed on 416 RCT images and 213 mast cell tumors images. In the test set, our algorithm exhibited an excellent classification performance in both RCT classification (accuracy: 91.66%) and mast cell tumors grading (accuracy: 100%). Misdiagnoses were encountered for histiocytomas in the train set and for melanomas in the test set. For mast cell tumors the reduction of a grade was observed in the train set, but not in the test set. To the best of our knowledge, the proposed model is the first fully automated algorithm in histological images specifically developed for veterinary medicine. Being very fast (average computational time 2.63 s), this algorithm paves the way for an automated and effective evaluation of canine tumors.

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Mast Cell Tumors and Histiocytomas in Domestic Goats and Diagnostic Utility of CD117/c-Kit and Iba1 Immunohistochemistry

Veterinary Pathology ◽

10.1177/0300985820988145 ◽

2021 ◽

Vol 58 (3) ◽

pp. 508-515

Author(s):

T. William O’Neill ◽

Christiane V. Löhr

Keyword(s):

Mast Cell ◽

Neuronal Cell ◽

Normal Lung ◽

Surgical Removal ◽

Round Cell ◽

Diagnostic Challenge ◽

Mast Cell Tumors ◽

Neuronal Cell Bodies ◽

Cutaneous Mast Cell ◽

Round Cell Tumors

Cutaneous round cell tumors in goats present a diagnostic challenge. In this article, we provide a description of caprine cutaneous mast cell tumors (MCT) and histiocytomas, and report on the validation of anti-human antibodies to CD117/KIT and Iba1 by immunohistochemistry on a range of caprine tissues. Cells immunolabeled for CD117/KIT included resident mast cells in normal lung and skin, interstitial cells of Cajal (intestine), and neuronal cell bodies (brain). Cells immunolabeled for Iba1 included resident macrophages in many tissues including normal lung, dendritic cells (hemolymphatic tissues), Kupffer cells, and microglia. Of 5 cutaneous MCT, only one had metachromasia of cytoplasmic granules; however, neoplastic cells of all 5 MCT had positive immunolabeling for CD117/KIT. The CD117/KIT immunolabeling pattern was predominately focal paranuclear in 3 cases, and cytoplasmic or membranous in 1 case each. Two histiocytomas were identified and had strong positive immunolabeling for Iba1 but not CD117/KIT. All 7 cutaneous round cell tumors described herein occurred in goats less than 4 years of age; the 2 cutaneous histiocytomas were in goats less than 14 months of age. Neither of the cutaneous histiocytomas recurred within 24 months of surgical removal.

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Histomorphological and immunohistochemical characterization of 172 cutaneous round cell tumours in dogs

Pesquisa Veterinária Brasileira ◽

10.1590/s0100-736x2012000800016 ◽

2012 ◽

Vol 32 (8) ◽

pp. 772-780 ◽

Cited By ~ 2

Author(s):

Marina Rios Araújo ◽

Ingred Sales Preis ◽

Gleidice Eunice Lavalle ◽

Geovanni Dantas Cassali ◽

Roselene Ecco

Keyword(s):

Mast Cell ◽

Plasma Cell ◽

Cell Lymphoma ◽

Morphological Characteristics ◽

B Cell Lymphoma ◽

Positive Staining ◽

Round Cell ◽

Histologic Diagnosis ◽

Cutaneous Lymphomas ◽

Formalin Fixed

This paper describes the use of a panel of antibodies (CD117, CD3, CD79a, CD45, cytokeratin, vimentin and E-cadherin) on formalin-fixed, paraffin-embedded sections of canine cutaneous round cell tumours. Neoplastic tumours were diagnosed by histology and histochemical stains and included 107 mast cell tumours, 31 cutaneous histiocytomas, two localized histiocytic sarcomas, 21 cutaneous lymphomas, three plasma cell tumours, one transmissible venereal tumour and seven unclassified round cell tumours. The histologic diagnosis was modified in 39.5% of the total 172 neoplasms. The staining for CD45 and Ecadherin were variable, and therefore, the final diagnoses of cutaneous histiocytoma and localized histiocytic sarcoma were made based on histology in association with negative results for CD3, CD79a, CD117 and cytokeratin. The cellular origin of unclassified round cell tumours was defined in all cases. Cutaneous B-cell lymphoma and plasma cell tumours were CD79a-positive and could be distinguished from each other by the morphological characteristics. Mast cell tumours and T cell lymphoma were CD117 and CD3 positive, respectively. The positive staining for vimentin and the negative staining for CD3, CD79a, CD117 and cytokeratin favoured the diagnosis of transmissible venereal tumours. Thus, the final diagnosis of cutaneous round cell tumours should be based on the interpretation of immunohistochemical results together with the cellular morphology observed by histology. Therefore, more studies to optimize the specific markers in formalin-fixed, paraffinembedded tissues (especially for histiocytes) are required for definitive diagnosis of round cell tumours in dogs.

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Histological and immunohistochemical practical studies of canine cutaneous tumors

Medycyna Weterynaryjna ◽

10.21521/mw.5558 ◽

2016 ◽

Vol 72 (9) ◽

pp. 571-579

Author(s):

Donatas Šimkus ◽

Saulius Petkevičius ◽

Gediminas Pridotkas ◽

Ligita Zorgevica-Pockeviča ◽

Viktoras Maskaliovas ◽

...

Keyword(s):

Epithelial Cells ◽

Toluidine Blue ◽

Fibrous Tissue ◽

Positive Staining ◽

Round Cell ◽

Mast Cell Tumors ◽

Melanocytic Tumors ◽

Subcutaneous Tissues ◽

Follicular Tumors ◽

Round Cell Tumors

A total of one hundred and fifty-three canine cutaneous tumors were examined and analyzed using the standard haematoxylin-eosin staining method. Additionally, tumors were examined immunohistochemically (41.4%) with antibodies LP34, AE1/AE3, V9 and histochemically (24.8%) with toluidine blue. Epithelial and melanocytic tumors of the skin accounted for 52.3% and mesenchymal tumours constituted 47.7%. All epidermal and follicular tumors demonstrated positive immunostaining for “LP34” antibodies. Fibromas and fibrosarcomas, which were immunohistochemically positive for antibodies “V9”, demonstrated negative immunostaining for antibodies “LP34”. As many as 47.4% of all round cell tumors showed positive staining with toluidine blue. Antibodies “LP34” are helpful for the differential diagnosis of epithelial cells of tumors in canine skin, skin adnexal and subcutaneous tissues. Antibodies “AE1/AE3” could be helpful for detecting metastatic glandular epithelial cells in the skin. Moreover, antibodies “V9” could be a useful tool to diagnose the cutaneous tumors consisting of fibrous tissue cells. Staining round cell tumors with toluidine blue may give valuable information on regarding mast cell tumors.

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IL-33 promotes MHC class II expression in murine mast cells

Journal of Dermatological Science ◽

10.1016/j.jdermsci.2016.08.226 ◽

2016 ◽

Vol 84 (1) ◽

pp. e73

Author(s):

Tomonobu Ito ◽

Chizu Egusa ◽

Tatsuo Maeda ◽

Nobuhiro Nakano ◽

Chiharu Nishiyama ◽

...

Keyword(s):

Mast Cells ◽

Mhc Class Ii ◽

Class Ii

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Effects of electrochemotherapy with cisplatin and peritumoral IL-12 gene electrotransfer on canine mast cell tumors: a histopathologic and immunohistochemical study

Radiology and Oncology ◽

10.1515/raon-2017-0035 ◽

2017 ◽

Vol 51 (3) ◽

pp. 286-294 ◽

Cited By ~ 14

Author(s):

Claudia Salvadori ◽

Tanja Svara ◽

Guido Rocchigiani ◽

Francesca Millanta ◽

Darja Pavlin ◽

...

Keyword(s):

Mast Cell ◽

T Lymphocytes ◽

Mhc Class Ii ◽

Cellular Response ◽

Histopathological Examination ◽

Combined Treatment ◽

Gene Electrotransfer ◽

Neoplastic Cells ◽

Ki 67 ◽

Mast Cell Tumors

AbstractBackgroundThe study was aimed to characterize tumor response after combined treatment employing electrochemotherapy with IL-12 gene electrotransfer in dogs with spontaneous mast cell tumors (MCT).Materials and methodsEleven dogs with eleven MCTswere included in the study. Histological changes were investigated in biopsy specimens collected before the treatment (T0), and 4 (T1) and 8 weeks (T2) later. Cellular infiltrates were characterized immunohistochemically by using anti CD3, CD20, Foxp3 (Treg), CD68 and anti MHC-class II antibodies. Proliferation and anti-apoptotic activity of neoplastic cells were assessed using anti Ki-67 and Bcl-2 antibodies. Angiogenetic processes were investigated immunohistochemically by using anti Factor VIII and anti CD31 antibodies and micro vessel density quantification.ResultsHistopathological examination of samples at T0confirmed the diagnosis and the presence of scanty infiltrates consisted mainly of T-lymphocytes and macrophages. At T1and T2neoplastic cells were drastically reduced in 7/11 cases, small clusters of neoplastic cells were detected in 3/11 cases and 1/11 cases neoplastic cells were still evident. Proliferation activity of neoplastic cells was significantly reduced at T1and T2and expression of anti-apoptotic protein at T1. Microvessel density was drastically reduced in all samples after treatment. The number of T-lymphocytes increased at T1, although not significant, while Treg were significant higher at T1and macrophages at T2.ConclusionsThe combined electrochemotherapy and IL-12 gene electrotransfer effectively induced a cellular response against neoplastic cells characterized mainly by the recruitment of T-lymphocytes and macrophages and a fibrotic proliferation with reduction of microvessels.

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