CCH cells are potent stimulators in the allogeneic mixed leucocyte reaction

Canine cutaneous histiocytoma (CCH) has been identified as a tumour of epidermal Langerhans cells (LCs) on the basis of immunophenotypic studies. Neoplastic Langerhans cells (CCH-LCs) were isolated from lesions of canine cutaneous histiocytoma. The CCH-LC cells expressed CD1b, CD11/18, CD45, MHC-I, and MHC-II. The CCH-LC cells were potent stimulators of the mixed leucocyte reaction (MLR) in vitro when compared to PBMCs from the tumour-bearing animals. This provides evidence that the neoplastic cells in CCH have functional as well as immunophenotypic characteristics of Langerhans cells.     

Utilization of cytoplasmic lysozyme immunoreactivity as a histiocytic marker in canine histiocytic disorders

Immunoreactive lysozyme was readily detectable in canine histiocytic disorders including systemic histiocytosis, malignant histiocytosis and granulomatous panniculitis. Lysozyme was less reliable as a histiocytic marker in cutaneous histiocytoma; forty percent of these tumors were negative for lysozyme expression. The marked heterogeneity in lysozyme expression in cutaneous histiocytoma may indicate that a proportion of these tumors show relatively primitive histiocytic differentiation and do not express lysozyme. Alternatively, this same proportion may exhibit a phenotype akin to cutaneous Langerhans cells which do not contain lysozyme. Lysozyme was not detectable in the tumor cells in lymphomatoid granulomatosis, atypical cutaneous histiocytoma, and histiocytic lymphosarcoma. Other evidence that these three disorders do not represent true histiocytic proliferative disorders is discussed.     

Immunohistochemical expression of E-cadherin does not distinguish canine cutaneous histiocytoma from other canine round cell tumors

Immunohistochemistry for E-cadherin (ECAD) has been used to distinguish canine cutaneous histiocytoma from other leukocytic neoplasms ("round cell tumors"). To determine the specificity of this test, 5 types of canine cutaneous round cell tumors were evaluated for immunohistochemical expression of ECAD. Tumors of all 5 types had variable cytoplasmic, plasma membrane, and/or paranuclear ECAD expression: All 13 cutaneous histiocytomas were ECAD+; all but 1 of 14 mast cell tumors expressed ECAD; 10 of 12 epitheliotropic lymphomas reacted with E-cadherin antibody; of 72 plasmacytomas, 54 were ECAD+; and 5 of 5 histiocytic sarcomas were positive. Conclusions based on these results include the following: First, immunoreactivity for ECAD is not limited to leukocytes of cutaneous histiocytoma; second, antibody to ECAD also labels neoplastic cells in most mast cell tumors, plasmacytomas, cutaneous histiocytic sarcomas, and epitheliotropic lymphomas; third, although most histiocytomas have membranous ECAD expression, the immunoreactivity varies among round cell tumors and is frequently concurrent in different cellular compartments; fourth, the distinctively paranuclear ECAD expression pattern in epitheliotropic lymphomas might distinguish them from other round cell tumors; and, fifth, ECAD should be used with other markers (eg, MUM1 for plasmacytomas, KIT for mast cell tumors, CD3 and CD79a for lymphomas) to distinguish among canine round cell tumors.     

Canine hemophagocytic histiocytic sarcoma: a proliferative disorder of CD11d+ macrophages

Histiocytic disorders of dogs include histiocytoma, localized histiocytic sarcoma (HS), disseminated HS (malignant histocytosis), and the reactive histiocytoses: cutaneous and systemic. A common element to these diseases is proliferation of dendritic cells (DC) of either Langerhans cell (epithelial DC) or interstitial DC lineage. In this report, 17 dogs with hemophagocytic HS are described. Breeds affected included Bernese Mountain Dog (6), Golden Retriever (4), Rottweiler (3), Labrador Retriever (2), a mixed-breed dog, and a Schnauzer, which were from 2.5 to 13 years old. The dogs presented with Coombs negative responsive anemia in 16/17 dogs (94%), thrombocytopenia in 15/17 dogs (88%), hypoalbuminemia in 16/17 dogs (94%), and hypocholesterolemia in 11/16 dogs (69%). All dogs died or were euthanized. The clinical course ranged from 2 to 32 weeks (mean 7.1 weeks). Diffuse splenomegaly with ill-defined masses was consistently present. Microscopic lesions were prevalent in spleen, liver, lung, and bone marrow. Metastasis occurred by insidious intravascular invasion with minimal mass formation. Histiocytes were markedly erythrophagocytic and accompanied by foci of extramedullary hemopoiesis. Cytologically, the histiocytes varied from well differentiated to atypical, with atypia more prevalent in spleen than bone marrow. These tumors arose from splenic red pulp and bone marrow macrophages, which expressed major histocompatibility complex class II and the beta2 integrin, CD11d. They had low and/or inconsistent expression of CD1 and CD11c, which are dominantly expressed by canine nonhemophagocytic HS of DC origin. Canine histiocytic proliferative diseases now encompass proliferation of all members of the myeloid histiocytic lineage: Langerhans cells, interstitial DC, and macrophages.     

Feline pulmonary Langerhans cell histiocytosis with multiorgan involvement

Histiocytic proliferative diseases are uncommon in cats, although recently a progressive histiocytosis of the skin with terminal involvement of internal organs has been described in cats. Here we describe 3 cats (2 males and 1 female) with pulmonary Langerhans cell histiocytosis (PLCH). The cats were euthanized due to progressive respiratory clinical symptoms and deterioration. Macroscopically, extensive, multifocal to confluent, pulmonary masses were evident. Infiltration of pancreas (2 cats), kidneys (1 cat), liver (1 cat), as well as tracheobronchial, hepatosplenic, or mesenteric lymph nodes (2 cats) was observed by gross or microscopic examination. The infiltrating cells had histiocytic morphology with cytologic atypia characterized by anisokaryosis and hyperchromasia regionally within infiltrated tissues. Lesional histiocytes expressed vimentin, CD18, and E-cadherin. Expression of E-cadherin was usually markedly reduced in extra-pulmonary lesions, which is consistent with possible down-regulation of E-cadherin associated with distant migration from the lung. Transmission electron microscopy demonstrated intracytoplasmic organelles consistent with Birbeck's granules of Langerhans cells in the lesional histiocytes in all cats, except in the pancreas of one cat. These findings were compatible PLCH with limited organ involvement of humans. It remains unproven whether feline PLCH represents a reactive or neoplastic cell proliferation.     

Resident Dendritic Cells in the Epidermis: Langerhans Cells, Merkel Cells and Melanocytes

Résumé— En plus des kératinocytes, l'épiderme contient des cellules résidentes de morphologie dendritique. Ce sont principalement la cellule de Langerhans, la cellule de Merkel et le mélanocyte. Ces cellules ont des fonctions diverses dans le tégument. Le mélanocyte assure la pigmentation cutanée et la protection contre les radiations U.V., et pourrait intervenir également dans la modulation de l'inflammation cutanée. La cellule de Langerhans intervient dans la surveillance immunologique des surfaces corporelles ecternes. La cellule de Merkel a des fonctions neuroendocrines. Cet article donne un aperçu de la structure et de la fonction de ces trois importantes populations cellulaires dans la peau. [Resident dentritic cells in the epidermis: Langerhans cells, Merkel cells and melanocytes (Cellules dentritiques résidentes de l'épiderme: cellules de Langerhans, cellules de Merkel et mélanocytes). Resumen— La epidermis contiene, además de los queratinocitos, células résidentes de morfologia dendria dendritica. Estas son principalmente las células de Langerhans, las células de Merkel y los melanocitos. Estas células tienen varias funciones en el integumento. Los melanocitos se encargan de la pigmentación y protección de la piel contra la radiación ultravioleta y tarrtbién participan en la regulación de la inflamación cutánea. Las células de Langerhans ayudan en la regulación inmunológica en la superficie externa. Las células de Merkel tienen funciones neuroendocrinas. Esta revisión da un repaso general a la estructura y función de tres importantes células de la piel. [Resident dendsuitic cells in the epidermis: Langerhans cells, Merkel cells and Melanocytes (Células résidentes en la epidermis: células de Langerhans, células de Merkel y melanocitos). Abstract— In addition to the keratinocytes, the epidermis contains resident cells of dendritic morphology. These are principally the Langerhans cell, Merkel cell and melanocyte. These cells have a number of different functions in the integument. The melanocyte is responsible for skin pigmentation and protection against UV radiation, and may also play a role in the modulation of cutaneous inflammation. The Langerhans cell aids in the immunological monitoring of the body's external surfaces. The Merkel cell has neuroendocrine functions. This review gives an overview of the structure and function of these three important cells of the skin.     

ISVD‐1 Highlights of the skin immune system

Similar to intestine and pulmonary tissue, skin is in contact with the environment and hence, requires efficient immune surveillance. The concept of skin‐associated lymphoid tissue (SALT) was first introduced in 1978. The skin immune system (SIS) with its cellular and humoral components and the associated draining lymph nodes fulfills the three major functions of the innate and adaptive immune system: (1) induction of the primary immune response; (2) expression of immunity to previously encountered antigens (memory); and (3) avoidance of deleterious immune responses to nonthreatening antigens (tolerance). The cellular components of the SIS include dendritic antigen‐presenting cells (Langerhans cells and dermal dendritic cells), skin‐homing αβ‐T lymphocytes and γδ‐T lymphocytes, keratinocytes and their environment, the microvascular unit (endothelial cells of the dermal postcapillary venules), neural cells and dendritic neural processes, and the draining lymph node with its high endothelial venules. Humoral components of the SIS are defensins, complement components, immunoglobulins, cytokines, chemokines, fibrinolysin, eicosanoids and neuropeptides. Important characteristics and functions of these components and their interactions with each other will be discussed with regard to normal skin versus recruitment phase, retention phase and resolution phase associated with inflammatory conditions. The discussion will primarily focus on cutaneous dendritic antigen‐presenting cells, cutaneous lymphocytes and their homing into the skin, and keratinocytes. Moreover, the importance of the microvascular unit in relation to leukocyte recruitment and migration will be discussed.     

Feline progressive histiocytosis

Histiocytic proliferative diseases include reactive and neoplastic proliferations of dendritic cells (DC) or macrophages. Various forms of DC proliferations have been documented in humans and dogs; their etiology is largely unknown. With the exception of a few case reports, histiocytic proliferations have not been characterized in cats. This study summarizes clinical, morphologic, and immunophenotypic features of a feline progressive histiocytosis (FPH) in 30 cats. There was no breed or age predilection. Females were more often affected than males. Solitary or multiple nonpruritic firm papules, nodules, and plaques had a predilection for feet, legs, and face. Lesions consisted of poorly circumscribed epitheliotropic (13/30) and nonepitheliotropic (17/30) histiocytic infiltrates of the superficial and deep dermis, with variable extension into the subcutis. The histiocytic population was relatively monomorphous early in the clinical course. With disease progression, cellular pleomorphism was more frequently encountered. Histiocytes expressed CD1a, CD1c, CD18, and major histocompatibility complex class II molecules. This immunophenotype suggests a DC origin of these lesions. Coexpression of E-cadherin, a feature of cutaneous Langerhans cells, was only observed in 3 cats. FPH followed a progressive clinical course; the lesions, however, were limited to the skin for an extended period of time. Terminal involvement of internal organs was documented in 7 cases. Treatment with chemotherapeutics or immunosuppressive and immunomodulatory drugs was not successful. The etiology of FPH remains unknown. FPH is best considered an initially indolent cutaneous neoplasm, which is mostly slowly progressive and may spread beyond the skin in the terminal stage.     

ISVD-2 Chytridiomycosis – an emerging and fatal infectious skin disease of amphibians

Similar to intestine and pulmonary tissue, skin is in contact with the environment and hence, requires efficient immune surveillance. The concept of skin-associated lymphoid tissue (SALT) was first introduced in 1978. The skin immune system (SIS) with its cellular and humoral components and the associated draining lymph nodes fulfills the three major functions of the innate and adaptive immune system: (1) induction of the primary immune response; (2) expression of immunity to previously encountered antigens (memory); and (3) avoidance of deleterious immune responses to nonthreatening antigens (tolerance). The cellular components of the SIS include dendritic antigen-presenting cells (Langerhans cells and dermal dendritic cells), skin-homing αβ-T lymphocytes and γδ-T lymphocytes, keratinocytes and their environment, the microvascular unit (endothelial cells of the dermal postcapillary venules), neural cells and dendritic neural processes, and the draining lymph node with its high endothelial venules. Humoral components of the SIS are defensins, complement components, immunoglobulins, cytokines, chemokines, fibrinolysin, eicosanoids and neuropeptides. Important characteristics and functions of these components and their interactions with each other will be discussed with regard to normal skin versus recruitment phase, retention phase and resolution phase associated with inflammatory conditions. The discussion will primarily focus on cutaneous dendritic antigen-presenting cells, cutaneous lymphocytes and their homing into the skin, and keratinocytes. Moreover, the importance of the microvascular unit in relation to leukocyte recruitment and migration will be discussed.     

Progressive Langerhans' cell histiocytosis in a puppy

A 8-month-old, female miniature Dachshund dog was presented for the complaint of pruritic, generalized, multiple nodules and plaques. Two months previously, a nodule on the left pinna was excised and diagnosed as a cutaneous histiocytoma. One month post-excision, a nodule reappeared at the same site and, shortly thereafter, additional nodules developed. Histopathological examination revealed a diffuse proliferation of histiocytic cells, which reacted strongly to antibodies for vimentin and lysozyme. Immunophenotypic analysis showed that most of the cells expressed CD1a, CD1c, CD11c, CD18, CD45 and MHC class II markers. Electron microscopic examination revealed cytoplasmic filopodia and paracrystalline structures. These findings indicated that the cells originated from Langerhans' cell. The disease progressed despite glucocorticoid therapy and griseofulvin was administered as an immunomodulating drug. All lesions resolved completely after 7 weeks of griseofulvin treatment. The dog, however, died three months later after discontinuation of griseofulvin therapy and a necropsy was not performed. It is considered that the present canine dermatosis corresponds with a severe form of Langerhans' cell histiocytosis in humans rather than canine cutaneous histiocytoma.     

Cytology of canine cutaneous round cell tumors. Mast cell tumor, histiocytoma, lymphosarcoma and transmissible venereal tumor.

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.     

A case of giant cell tumor of soft parts in a horse

A 12‐year‐old British Warmblood mare was examined by the referring veterinarian for evaluation of a cutaneous lesion on the dorsal thorax to the right of the midline. Cytologic examination of fine‐needle aspirates from the mass was supportive of a giant cell tumor of soft parts (GCTSP). Laser surgical excision and postoperative methyl aminolevulinate (MAL) photodynamic therapy (PDT) were performed. Histologic examination of the mass confirmed the cytologic diagnosis. At 8 months from surgery, no evidence of recurrence has been observed. Giant cell tumors of soft parts are rare cutaneous neoplasms, observed in several domestic species, including the horse where they commonly appear as superficial cutaneous lesions without aggressive biologic behavior. Previously classified as giant cell variant of malignant fibrous histiocytoma, these superficial tumors have now been designated as giant cell tumors of soft tissue or giant cell tumors of low malignant potential within the category of fibrohistiocytic neoplasms.     

Mast cell tumour and cutaneous histiocytoma excision wound healing in general practice

Objective : To assess whether wounds from incomplete mast cell tumour excisions are at greater risk of healing complications than wounds from complete excisions, or cutaneous histiocytomas. Methods : Mast cell tumours and cutaneous histiocytomas submitted to Nationwide Laboratories between November 1, 2007 and April 30, 2008 were selected. Questionnaires were sent to submitting veterinarians requesting details of tumour characteristics, clinical approach to the tumour and wound healing. Results : Three hundred and eighty‐six mast cell tumours and 524 cutaneous histiocytomas were identified. One hundred and eighty‐five mast cell tumours and 244 cutaneous histiocytomas questionnaires were returned (47% response). Wound complications arose in 20% of mast cell tumours and 21% of cutaneous histiocytomas. Multivariable analysis confirmed that larger tumours, tumours on the feet and a soft/“baggy” appearance, were significantly associated with a greater frequency of problems, leading to delayed wound healing and dehiscence. Clinical Significance : Incomplete mast cell tumour excision does not lead to greater risk of wound complications. Mast cell tumour surgical wounds have a similar rate of wound complications as cutaneous histiocytoma wounds.     

In vivo confocal microscopy of the normal equine cornea and limbus

Objective  To describe morphologic features, pachymetry and endothelial cell density of the normal equine cornea and limbus by in vivo confocal microscopy.
Animals studied  Ten horses without ocular disease.
Procedure  The central and peripheral corneas were examined with a modified Heidelberg Retina Tomograph II and Rostock Cornea Module using a combination of automated and manual image acquisition modes. Thickness measurements of various corneal layers were performed and endothelial cell density determined.
Results  Images of the constituent cellular and noncellular elements of the corneal epithelium, stroma, endothelium, and limbus were acquired in all horses. Corneal stromal nerves, the subepithelial nerve plexus, and the sub-basal nerve plexus were visualized. Cells with an appearance characteristic of Langerhans cells and corneal stromal dendritic cells were consistently detected in the corneal basal epithelium and anterior stroma, respectively. Median central total corneal thickness was 835 μm (range 725–920 μm) and median central corneal epithelial thickness was 131 μm (range 115–141 μm). Median central endothelial cell density was 3002 cells per mm² (range 2473–3581 cells per mm²).
Conclusions  In vivo corneal confocal microscopy provides a noninvasive method of assessing normal equine corneal structure at the cellular level and is a precise technique for corneal sublayer pachymetry and cell density measurements. A resident population of presumed Langerhans cells and corneal stromal dendritic cells was detected in the normal equine cornea. The described techniques can be applied to diagnostic evaluation of corneal alternations associated with disease and have broad clinical and research applications in the horse.     

Multiple cutaneous histiocytomas in a cat

A 15-year-old domestic long haired cat developed nodular cutaneous masses over the right shoulder that were removed surgically. Similar masses developed in multiple cutaneous sites over the following three years. In each case, the lesions were characterized by diffuse dermal infiltration by histiocytic cells with a low mitotic rate and evidence of epidermotropism. The neoplastic population uniformly expressed class II molecules of the Major Histocompatibility Complex, but did not express the T and B lymphoid markers CD3 and CD79. Perivascular aggregates of CD3⁺ T lymphocytes were located at the deep margins of the tumour nodules. The clinical, histopathological and immunohistochemical features of these tumours are consistent with cutaneous histiocytoma. This tumour has not been previously well-documented in the cat.     

Multiple, resolving, cutaneous histiocytoma in a dog

Mycosis fungoides was initially diagnosed in a 7.5-year-old German Shepherd Dog with generalized canine cutaneous histiocytoma. Lesions resolved without treatment over approximately 16 weeks. The final diagnosis of histiocytoma with 2 histopathologic patterns was obtained by use of a special staining technique for the detection of lysozyme found in histiocytes.     

P-66 Disseminated histiocytic sarcoma in a dog receiving long-term immunosuppressive therapy

Canine cutaneous histiocytoma is a common tumor of young dogs. Nodules are single or multiple, alopecic and well demarcated. We studied 15 young dogs with cutaneous histiocytoma. Cutaneous nodules were cytologically and histologically investigated using routine techniques. The subject for morphometry was the nuclear area of histiocytes. The figures were statistically interpreted (Student's t- test). Cytologically, histiocytes represented the largest population, being large-sized cells. The nucleus:cytoplasm ratio was large with oval or bean-shaped nuclei and a fine arrangement of chromatin. The cytoplasm was discrete and basophilic, or abundant and foamy. Five regressive stages were histologically described. Stage 1 was represented by dermal and hypodermal histiocyte proliferation, covered by an intact, uninfiltrated and atrophic epidermis. Epidermal ulcers and necrotizing foci with neutrophilic, lymphocytic and plasma cell infiltrations occurred in Stages 2, 3 and 4. Stage 5 looked like a chronic dermatitis. In Stage 1, the nuclear area was the smallest (4615 ± 62 pixels) with a small coefficient of variation (14.28%), proving a homogeneous population. Stage 2 histiocytes had a larger and more heterogeneous nuclear area (6116 ± 130 pixels) with a 21.12% coefficient of variation. In Stage 3, the average nuclear area (6431 ± 115 pixels) was larger than that for Stage 2 and had a 19.18% coefficient of variation. There were no significant differences between the nuclear areas of histiocytes from Stages 2 and 3; however, significant differences occurred between Stages 1 and 2, and between Stages 1 and 3.
Funding: Self-funded .     

Cutaneous histiocytosis in dogs

Multifocal cutaneous histiocytic lesions were recognized in 9 dogs. Clinically, the dogs had multiple erythematous plaques or nodules in the skin (1 to 5 cm diameter). Histologically, the lesions were comprised of dermal or pannicular infiltrates of large histiocytic cells, with varying numbers of other inflammatory cells intermixed. By electron microscopy, the cells resembled those of canine cutaneous histiocytoma. The lesions seemed to wax and wane and appeared in new sites, regardless of treatment. The dogs ranged in age from 2 to 13 years; 7 dogs were under 6 years of age. Both sexes and various breeds were represented. An infectious agent could not be identified.     

Simulated solar UVB exposure inhibits transcutaneous immunization to cholera toxin via an irradiated skin site in cattle

Transcutaneous immunization (TCI) is a new needle-free vaccination technology with the potential to reduce the risk of needle-borne disease transmission and carcass damage within the livestock industries. The principal antigen-presenting cell involved in TCI is thought to be the epidermal Langerhans cell. Langerhans cell function is inhibited by cutaneous ultraviolet-B radiation (UVB) exposure. Such exposure may inhibit TCI through sun exposed skin sites due to the phenomenon of local low dose photoimmunosuppression. TCI of cattle to cholera toxin (CT) resulted in the generation of a serum anti-CT-specific IgG(2) response. However, exposure of cattle to a sub-inflammatory dose of simulated solar UVB (2.43 x 10(3)J/m(2)) significantly (P<0.05) inhibited TCI to CT via irradiated skin sites.     

P‐65 Canine cutaneous histiocytoma: morphology and morphometry

Canine cutaneous histiocytoma is a common tumor of young dogs. Nodules are single or multiple, alopecic and well demarcated. We studied 15 young dogs with cutaneous histiocytoma. Cutaneous nodules were cytologically and histologically investigated using routine techniques. The subject for morphometry was the nuclear area of histiocytes. The figures were statistically interpreted (Student's t‐test). Cytologically, histiocytes represented the largest population, being large‐sized cells. The nucleus:cytoplasm ratio was large with oval or bean‐shaped nuclei and a fine arrangement of chromatin. The cytoplasm was discrete and basophilic, or abundant and foamy. Five regressive stages were histologically described. Stage 1 was represented by dermal and hypodermal histiocyte proliferation, covered by an intact, uninfiltrated and atrophic epidermis. Epidermal ulcers and necrotizing foci with neutrophilic, lymphocytic and plasma cell infiltrations occurred in Stages 2, 3 and 4. Stage 5 looked like a chronic dermatitis. In Stage 1, the nuclear area was the smallest (4615 ± 62 pixels) with a small coefficient of variation (14.28%), proving a homogeneous population. Stage 2 histiocytes had a larger and more heterogeneous nuclear area (6116 ± 130 pixels) with a 21.12% coefficient of variation. In Stage 3, the average nuclear area (6431 ± 115 pixels) was larger than that for Stage 2 and had a 19.18% coefficient of variation. There were no significant differences between the nuclear areas of histiocytes from Stages 2 and 3; however, significant differences occurred between Stages 1 and 2, and between Stages 1 and 3. Funding: Self‐funded.