Verena K. Affolter

Localized and disseminated histiocytic sarcoma of dendritic cell origin in dogs.

Canine histiocytic proliferative disorders include a wide spectrum of diseases characterized by different biologic behaviors. The etiology and pathogenesis of these diseases are largely unknown. The clinicopathologic, morphologic and immunophenotypic characteristics of canine localized and disseminated histiocytic sarcoma were examined in 39 dogs. Rottweilers, Bernese Mountain Dogs, and retrievers were most commonly affected (79%). Localized histiocytic sarcomas (19 dogs) arose from a single site, and metastatic lesions were observed in draining lymph nodes. Predilection sites were subcutis and underlying tissues on extremities, but tumors occurred in other locations, including spleen, lung, brain, nasal cavity, and bone marrow. Disseminated histiocytic sarcomas (20 dogs), a multisystem disease previously described as malignant histiocytosis, primarily affected spleen, lungs, bone marrow, liver, and lymph nodes. Both localized and disseminated canine histiocytic sarcomas were composed of pleomorphic tumor cell populations. CD1+, CD4-, CD11c+, CD11d-, MHC II+, ICAM-1+, Thy-1± tumor cells were identified in all snap-frozen samples (31 dogs). This phenotype is characteristic for myeloid dendritic antigen-presenting cell lineage. Hence, canine localized and disseminated histiocytic sarcomas are likely myeloid dendritic cell sarcomas. Dendritic antigen-presenting cells are a heterogeneous cell population with regards to their ontogeny, phenotype, function, and localization. The exact sublineage of the proliferating dendritic antigen-presenting cells involved in canine histiocytic sarcomas remains to be determined. Phenotypic analysis of formalin-fixed tissues from eight dogs was limited by available markers. Morphologic features and the phenotype CD18+, CD3-, and CD79a- were the most useful criteria to indicate likely histiocytic origin.

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 β2 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.

Langerhans cell hyperplasia and IgE expression in canine atopic dermatitis.

Langerhans cells appear to be critical for IgE-mediated allergen capture and presentation in human atopic dermatitis. The present study sought to determine whether epidermal (i.e Langerhans cells) and dermal dendritic cells in the skin of dogs with atopic dermatitis are hyperplastic and expressed surface IgE. Frozen sections of lesional or nonlesional atopic and normal control canine skin were immunostained with CD1a-, CD1c-, and IgE-specific monoclonal antibodies. The enumeration of cells was performed by morphometry in both the epidermis and the dermis. Cell counts were compared with each individual’s total serum IgE levels. Higher numbers of epidermal and dermal dendritic cells were present in atopic dogs than in normal control animals. Epidermal Langerhans cell counts were significantly higher in lesional than in nonlesional atopic specimens. IgE+ dendritic cells were observed in lesional atopic epidermis and dermis, and nonlesional atopic dermis, but not in normal control skin specimens. The percentages of IgE+ dendritic cells were correlated with each patient’s total serum IgE levels. These results demonstrate dendritic cell hyperplasia and IgE expression in canine atopic dermatitis. Increased epidermal Langerhans cell counts in lesional specimens suggest an epidermal allergen contact in canine atopic dermatitis.

Canine cutaneous and systemic histiocytosis: Reactive histiocytosis of dermal dendritic cells

Canine histiocytic proliferative disorders include reactive diseases such as cutaneous and systemic histiocytosis and neoplastic diseases such as cutaneous histiocytoma and localized and disseminated histiocytic sarcoma (malignant histiocytosis). Their etiology and pathogenesis are unknown. Canine cutaneous and systemic histiocytosis target the skin and subcutis and have similar clinical behavior. Systemic histiocytosis also affects other organ systems. Clinicopathologic and phenotypic features of canine cutaneous and systemic histiocytosis were examined in this study. Canine cutaneous (18 cases) and systemic (26 cases) histiocytosis were characterized by angiocentric, pleocellular accumulations consisting of CD1+, CD11c+, MHCII+, CD4+, and Thy-1+ (CD90) activated dermal dendritic antigen-presenting cells (APC) with admixed CD3+, CD8+, TCRalphabeta+ T lymphocytes, and neutrophils. Hence, canine cutaneous and systemic histiocytosis represent two clinical manifestations of a reactive proliferation of dermal dendritic cells. Cultures and special stains failed to identify infectious agents. Canine reactive histiocytoses respond to immunosuppressive therapy (cyclosporine A or leflunomide). Therefore, immune-dysregulatory mechanisms are likely to be involved. Spontaneous reactive histiocytoses are frequently seen in dogs, and they constitute an excellent model to study pathologic mechanisms involved in reactive proliferations of dermal dendritic APC.

Intradermal injections of equine allogeneic umbilical cord-derived mesenchymal stem cells are well tolerated and do not elicit immediate or delayed hypersensitivity reactions

BACKGROUND AIMS. The use of allogeneic mesenchymal stem cells (MSC) to treat acute equine lesions would greatly expand equine cellular therapy options; however, the safety and antigenicity of these cells have not been well-studied. We hypothesized that equine allogeneic umbilical cord tissue (UCT)-derived MSC would not elicit acute graft rejection or a delayed-type hypersensitivity response when injected intradermally. METHODS. Six Quarterhorse yearlings received 12 intradermal injections (autologous MSC, allogeneic MSC, positive control and negative control, in triplicate) followed by the same series of 12 injections, 3-4 weeks later, at another site. Wheals were measured and palpated at 0.25, 4, 24, 48, 72 h and 7 days post-injection. Biopsies were obtained at 48 and 72 h and 7 days post-injection. Mixed leukocyte reactions were performed 1 week prior to the first injections and 3 weeks after the second injections. RESULTS. There were no adverse local or systemic responses to two intradermal injections of allogeneic MSC. MSC injection resulted in minor wheal formation, characterized by mild dermatitis, dermal edema and endothelial hyperplasia, that fully resolved by 48-72 h. No differences were noted between allogeneic and autologous MSC. The second injection of MSC did not elicit more significant physical or histomorphologic alterations compared with the first MSC injection. Neither allogeneic nor autologous UCT-derived MSC stimulated or suppressed baseline T-cell proliferation in vitro prior to or after two MSC administrations. CONCLUSIONS. Equine allogeneic UCT MSC may be safely administered intradermally on multiple occasions without eliciting a measurable cellular immune response.

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.

Sporotrichosis: a retrospective evaluation of 23 cases seen in northern California (1987-2007).

Sporotrichosis is an uncommon to rare cutaneous and subcutaneous mycosis of animals and humans caused by the dimorphic fungus Sporothrix schenckii. Twenty-three mammalian cases of sporotrichosis examined between 1987 and 2007 at the University of California, Davis - Veterinary Medical Teaching Hospital, were retrospectively evaluated with regard to the historical, clinical, diagnostic and treatment findings. Cats were the most common species affected (n = 14). In addition, sporotrichosis was diagnosed in four dogs, four horses and a donkey. Six of 23 cases were diagnosed with the localized cutaneous form of sporotrichosis (four cats, one dog, one horse), 10 with the cutaneous-lymphatic form (four cats, two dogs, three horses and a donkey), and seven with the disseminated form (six cats, one dog). Two of 23 cases did not have skin lesions at the time of diagnosis (one cat, one dog). The most common mode of diagnosis was demonstration of S. schenckii on histopathological evaluation of tissue. In contrast with most previously described sporotrichosis infections in cats, few to no fungal organisms were seen in histopathological samples (haematoxylin and eosin and special stains) in five of the 14 cats. Treatments received included itraconazole (12 cats, one dog), ketoconazole (three dogs), fluconazole (one cat, one donkey), sodium iodide (four horses, one cat) and potassium iodide (one cat, one horse, one donkey). The prognosis for successful treatment was good in all species. Fluconazole was successful in inducing resolution of the cutaneous lesions and controlling the infection in one cat with disseminated sporotrichosis.

Canine epitheliotropic cutaneous T-cell lymphoma: an investigation of T-cell receptor immunophenotype, lesion topography and molecular clonality

Canine epitheliotropic cutaneous T-cell lymphoma (CTCL) is a spontaneous neoplasm of the skin and mucous membranes of aged dogs. The WHO classification of tumours of haematopoietic and lymphoid tissues in human beings recognizes three forms of cutaneous epitheliotropic CTCL: mycosis fungoides (MF), Sézary syndrome and pagetoid reticulosis. In this series of dogs (n = 56), there were 39 cases of MF, 16 cases of pagetoid reticulosis and a single case of Sézary syndrome. Epitheliotropic T cells in CTCL lesions expressed CD8 in 44 of 55 dogs (80%) assessed; neither CD4 nor CD8 was expressed in the remainder. This contrasts with human MF in which alphabeta T-cell receptors (TCR) and CD4 are dominantly expressed. Molecular clonality assessment of canine epitheliotropic CTCL utilizing PCR primers specific for canine TCR gamma (TCRG) was performed. Of the 45 canine cases assessed, TCRG monoclonality was detected in 36 cases (80%). TCR typing of canine epitheliotropic CTCL revealed that TCRgammadelta was expressed in 60% of cases, including all cases of canine pagetoid reticulosis assessed. Either muco-cutaneous junctions or tissues of the oral cavity were the sites of lesions in 32 dogs (57%) with epitheliotropic CTCL. Analysis of the topography of lesions revealed an association with TCR type. If epitheliotropic CTCL lesions occurred in both locations, T cells were more likely to express TCRgammadelta (gammadelta : alphabeta = 2.0). These data establish that canine skin trafficking T cells have a far wider range than previously thought; this includes tongue, gingival, buccal and palatine mucosae.

Activity, Expression and Genetic Variation of Canine β-Defensin 103: A Multifunctional Antimicrobial Peptide in the Skin of Domestic Dogs

The skin functions as more than a physical barrier to infection. Epithelial cells of the skin can synthesize antimicrobial peptides, including defensins, which exhibit direct antimicrobial activity. Here we characterize the expression pattern, genetic variation and activity of the major β-defensin expressed in canine skin, canine β-defensin 103 (CBD103). The gene encoding CBD103 exhibits two forms of polymorphism: a common 3-basepair deletion allele and a gene copy-number variation. Golden retrievers and Labrador retrievers were the only breeds that encoded the variant allele of CBD103, termed CBD103ΔG23. Both these breeds also exhibited a CBD103 gene copy-number polymorphism that ranged from 2 to 4 gene-copies per diploid genome. Recombinant CBD103 and CBD103ΔG23, as well as the human ortholog human β-defensin 3 (hBD3) and hBD3ΔG23, showed potent and comparable antimicrobial killing against both methicillin-susceptible and methicillin-resistant Staphylococcus pseudintermedius. Skin biopsy specimens from dogs with atopic dermatitis revealed CBD103 expression levels similar to those in healthy controls and comparable at lesional and nonlesional sites. This expression pattern in dogs differs from the previously reported reduced expression of the human ortholog in atopic dermatitis. Overall, the similarities of CBD103 and its human ortholog reported here support the notion that the domestic dog may serve as a valuable model for studying β-defensin biology in the skin.

Progressive Swelling, Hyperkeratosis, and Fibrosis of Distal Limbs in Clydesdales, Shires, and Belgian Draft Horses, Suggestive of Primary Lymphedema

BACKGROUND A condition characterized by progressive swelling, hyperkeratosis, and fibrosis of the distal limbs has been recognized in Shire, Clydesdale, and Belgian draft horses. This chronic progressive disease starts at an early age, progresses throughout the life of the horse, and often ends in disfigurement and disability of the limbs that inevitably leads to the horses premature death. This study was undertaken to better characterize this disease. METHODS AND RESULTS Six affected horses were donated for diagnostic workup. A detailed clinical, radiologic, gross, and histologic description is given in this report. The lesions in the limb consisted of progressive development of thick-walled lymphatics, associated with chronic dermal edema, inflammation, fibrosis, neovascularization, and elastin degeneration. In the end stages, arteriosclerosis and fibrosed veins were also present. The clinical signs and pathologic changes in this disease closely resemble the human condition of elephantiasis nostras verrucosa, a state in which chronic lymphedema plays a pivotal pathogenic role.