Immunoperoxidase staining for the detection of autoantibodies in canine autoimmune skin disease; comparison to immunofluorescence results

Skin sections from 22 dogs with autoimmune skin disease were stained with anti-canine IgG, IgM and IgA using an immunobridge immunoperoxidase method. Eight cases of lupus erythematosus, three cases of pemphigus vulgaris, and 11 cases of pemphigus foliaceus were included. Results of previously performed, direct immunofluorescence tests for the detection of canine immunoglobulin on skin were available on 17/22 cases. The immunoperoxidase method yielded an overall positive result in 59% (5/8 lupus erythematosus, 2/3 pemphigus vulgaris and 6/11 pemphigus foliaceus) versus an overall positive result of 47% for direct immunofluorescence (3/5 lupus erythematosus, 2/2 pemphigus vulgaris and 2/10 pemphigus foliaceus). The immunobridge immunoperoxidase method compared favorably to direct immunofluorescence testing of canine skin for autoantibody in cases of lupus erythematosis and pemphigus vulgaris, and was superior in cases of pemphigus foliaceus. This method should prove useful as an aid in the diagnosis of canine autoimmune skin disease.     

A review of autoimmune skin diseases in domestic animals: I - superficial pemphigus

In humans, the pemphigus denomination encompasses a group of autoimmune blistering skin diseases with intraepidermal separation resulting from cell-cell detachment by acantholysis. Entities are classified based on the level of blistering in the epidermis, and both superficial (pemphigus foliaceus, IgA pemphigus) and deep (pemphigus vulgaris, pemphigus vegetans and paraneoplastic pemphigus) variants are recognized. In domestic animals, subsets of pemphigus have been recognized since the mid-1970s, and the disease classification resembles that used for human patients. This article reviews up-to-date knowledge on the epidemiology, clinical signs, histopathology, immunopathology and treatment outcome of superficial pemphigus in domestic animals. Detailed information on canine, feline, equine and caprine pemphigus foliaceus, canine and feline pemphigus erythematosus and canine panepidermal pustular pemphigus is provided.     

Ultrastructural localization of pemphigus vulgaris antigen on canine keratinocytes in vivo and in vitro

Pemphigus antigens were localized, by use fo immunoelectron microscopy, on canine keratinocytes in vivo on esophageal mucosa and in vitro on established cultured keratinocytes. Convalescent sera from a human being with pemphigus vulgaris and a human being with pemphigus foliaceus reacted with the interdesmosomal cytoplasmic keratinocyte membrane of canine esophagus. Cultured canine keratinocytes expressed the pemphigus vulgaris antigen in a similar pattern, but did not carry the pemphigus foliaceus antigen. The differential presence of cell surface antigens and its relation to various forms of the disease are discussed.     

Prolonged remission after immunosuppressive therapy in six dogs with pemphigus foliaceus

Limited information is available on the long-term outcome of treatment of pemphigus foliaceus in dogs. The purpose of this study is to report that a prolonged remission can occur after discontinuation of immunosuppressive regimens in some animals with this disease. Six dogs were diagnosed with pemphigus foliaceus based on suggestive clinical signs and histopathology. These patients were treated either with immunosuppressive doses of oral glucocorticoids or with a combination of oral glucocorticoids and azathioprine. After clinical signs underwent complete remission, which occurred 1.5-5 months after immunosuppression was initiated, the drugs were tapered progressively and eventually withdrawn. The total duration of immunosuppressive therapy varied between 3 and 22 months. Skin lesions of pemphigus foliaceus did not recur for 1.5-6 years after treatment was stopped. These observations suggest that, in some dogs with pemphigus foliaceus, immunosuppression can lead to long-term remission of skin lesions, and that discontinuation of treatment is not necessarily followed by a recurrence of clinical signs.     

Immunologic diseases

Selected equine cutaneous diseases for which there is strong evidence of an immunological aetiology are discussed in this section. The reactive disorders of urticaria, Culicoides hypersensitivity, erythema multiforme, drug eruptions and systemic granulomatous disease are included. The autoimmune diseases of the horse with cutaneous manifestations (pemphigus foliaceus, pemphigus vulgaris, bullous pemphigoid and cutaneous lupus erythematosus) are discussed. The clinical signs and diagnostic features of these immunologically induced equine dermatoses are defined, and the challenges in making a definitive diagnosis and managing the clinical signs are confirmed.     

Altered immunohistochemical staining for desmoglein in skin biopsies in canine pemphigus foliaceus.

In this study, 50 cases of canine pemphigus foliaceus and 49 cases of canine superficial pyoderma were examined by immunohistochemical staining for patterns of desmoglein expression. In 31/50 (62%) of pemphigus foliaceus cases, there was an altered staining pattern for desmoglein consisting of distinct clumped deposits at the periphery of keratinocytes and/or dark cytoplasmic staining of acantholytic cells (consistent with internalization of desmoglein). In contrast, desmoglein staining in biopsies from cases of superficial pyoderma was diffusely pale without evidence for clumping or distinct internalization. This study demonstrates that epidermal desmoglein expression is altered in some cases of pemphigus foliaceus in dogs and suggests that immunohistochemical staining for this protein may be useful in diagnosis.     

Clinical and histopathological features of pemphigus foliaceus with and without eosinophilic infiltrates: a retrospective evaluation of 40 dogs

The importance of cellular infiltrates in tissues has been investigated as a diagnostic tool, mechanism of pathogenesis, and prognostic indicator in certain human diseases. Eosinophils, in particular, have a distinct role in the development of cutaneous lesions in human autoimmune diseases. Identification of an eosinophilic infiltrate can aid the diagnosis of immunobullous disease in the early stages of the disease process. In canine pemphigus foliaceus, eosinophils are present to a variable degree within lesional tissue. This study retrospectively evaluated 40 dogs with pemphigus foliaceus, and examined clinical and histologic features and final outcomes in cases with and without eosinophilic infiltrates. Twenty‐five of 40 dogs (63%) had an eosinophilic infiltrate in either the pustules/crust, follicular infundibulum or dermis. There was no statistically significant difference in clinical distribution or appearance of dermatological lesions, response to treatment, or disease outcome in dogs with or without an eosinophilic infiltrate. However, dogs with concurrent disease were significantly more likely to have an eosinophilic infiltrate (P = 0.01). Dogs with adverse effects associated with immunosuppressive therapy were significantly more likely to have an eosinophilic infiltrate (P = 0.05). Fifteen of 40 dogs (38%) had a history of allergic disease and a significantly higher proportion of these dogs had an eosinophilic infiltrate (P = 0.04). An eosinophilic infiltrate was found in more than half of the dogs in this study. These findings justify further studies to investigate the role of eosinophils in the pathogenesis, therapy and prognosis in dogs with pemphigus foliaceus.     

Diabetes mellitus induced in a dog after administration of corticosteroids and methylprednisolone pulse therapy

An 8-year-old ovariohysterectomized Chow Chow was referred because of dermatologic lesions diagnosed as pemphigus foliaceus. Intolerance to orally administered corticosteroids necessitated the use of methylprednisolone pulse therapy. One week after treatment, diabetes mellitus was diagnosed on the basis of blood and urine test results. For 3 years after treatment, the dog has remained a well-regulated diabetic. Complete remission of pemphigus foliaceus is maintained by alternate-day, orally administered prednisone (0.5 mg/kg of body weight).     

Comorbidity of ectopic thymoma-associated exfoliative dermatitis and pemphigus foliaceus in a cat

An 8-year-old cat was presented with pruritus, purulent paronychia, scaling, crusting, and spontaneous alopecia. Histopathology revealed intraepidermal neutrophilic pustular acantholytic dermatitis and hyperkeratotic cytotoxic interface dermatitis. No thoracic mass was seen on a lateral radiograph. Ectopic thymoma was discovered on necropsy. This case highlights the necessity for thorough investigation of any case of feline exfoliative dermatitis and pemphigus foliaceus for the presence of thymoma.Key clinical message:Comorbidity of exfoliative dermatitis and pemphigus foliaceus in a cat should prompt a thorough investigation for presence of a thymoma, possibly with advanced imaging techniques.     

Pemphigus foliaceus of the footpads in three dogs

Severe hyperkeratinization and villous hypertrophy of the footpads were seen in 3 middle-aged dogs. Peeling, fissuring, swelling, and ulcerations were noted on the margins of severely affected pads. Pain was evident in palpation and ambulation. Lesions were compatible with the traditional diagnosis of "hard pad disease". Histopathologic findings were diagnostic for canine pemphigus foliaceus in all 3 dogs, and direct immunofluorescence in an intercellular pattern was seen in both dogs that were tested. All 3 dogs responded to immunosuppressive dosages of corticosteroids.     

The longevity of immunoglobulin preservation in canine skin utilizing Michel's fixative

Skin biopsy specimens from 7 dogs with immune-mediated skin diseases diagnosed by routine histology and 5 dogs with other skin diseases were placed in Michel's transport medium for 4 to 9 years. Direct immunofluorescence yielded positive results in tissue samples from 3 dogs with pemphigus foliaceus and 2 dogs with discoid lupus erythematosus. Direct immunofluorescence was not seen in tissue samples from 1 dog with pemphigus foliaceus and 5 dogs with non immune-mediated skin diseases. Direct immunofluorescence was seen in skin biopsy specimens maintained in Michel's medium for 4 to 8 years.     

Use of tetracycline and niacinamide for treatment of autoimmune skin disease in 31 dogs.

A combination of niacinamide and tetracycline was used to treat 31 dogs with various autoimmune skin diseases (discoid lupus erythematosus, pemphigus foliaceus, pemphigus erythematosus, and bullous pemphigoid). Of the 20 dogs with discoid lupus erythematosus, 70% had excellent or good response to treatment. Serious side effects were not noticed in any dog.     

Pemphigus: current therapy

Pemphigus is an autoimmune skin disease that can present in a variety of forms and can be a challenging disease to manage and treat. An overview of the different forms of pemphigus and diagnostics are discussed including pemphigus foliaceus (PF), pemphigus erythematosus (PE), panepidermal pustular pemphigus (PPP), pemphigus vulgaris (PV) and paraneoplastic pemphigus (PNP). Emphasis on therapy is presented. Included are the most current commonly used therapeutics (glucocorticoids, azathioprine, chlorambucil and tetracycline and niacinamide); current alternative therapeutics (cyclosporin and tacrolimus and mycophenolate mofetil) and additional alternative therapeutics (cyclophosphamide, chrysotherapy, dapsone, sulfasalazine and intravenous immunoglobulin (IVIG) therapy).     

Two cases of feline leishmaniosis in Switzerland

Two cats with Leishmania species infections were investigated. The first had been imported from Spain with a non-healing, ulcerated nodule on a hindleg. The presence of Leishmania species was detected by histopathology and pcr on samples of skin. The lesion was unresponsive to treatment with allopurinol for three months but the cat was treated successfully by removing the lesion surgically. The second cat had lived in both Spain and Switzerland, and had a history of recurrent skin lesions on its head and neck. A diagnosis of pemphigus foliaceus was made on the basis of histopathology, but Leishmania species serology (elisa) and pcr of skin were positive, leading to a diagnosis of a Leishmania species infection combined with pemphigus foliaceus.     

Pemphigus foliaceus in the horse: a retrospective study of 20 cases

Twenty horses with pemphigus foliaceus were seen over a period of 15 years in a veterinary medical teaching hospital. Breeds seen were seven quarterhorses, five thoroughbreds, three cross-bred horses, two Arabians and one of each of the following: standardbred, Tennessee walker and warmblood. There was no breed, age or sex predisposition. Nine were mares, ten were geldings and one was a stallion. Ages ranged from 2.5 months to 25 years, with a mean of 8.6 years. Sixteen (80%) of the pemphigus foliaceus horses first exhibited signs between September and February. There was a statistically significant more common occurrence of pemphigus foliaceus during those months. Signs in the four other horses were first noted in March, May or June. Three of those horses were < 13 months of age. Oedema (14/20) and crusts (13/20) were the most common lesions. Pain was present in 9/20 horses, pruritus in 7/20 and pyrexia in 7/20. Follow-up was available for 13 horses. Five of these horses were euthanased. In three horses the reason for euthanasia was laminitis secondary to treatment. Four horses remained lesion-free after medication was discontinued. Two horses required maintenance medication and are doing well at the time of writing.     

Complement immunofluorescence in sera of dogs with pemphigus foliaceus

Sera from 14 dogs with active pemphigus foliaceus were tested for the presence of circulating antiepithelial antibodies. These antibodies could not be detected in the dogs with indirect immunofluorescent staining or in vitro complement-staining methods.     

Pemphigus in the dog: comparison of immunofluorescence and immunoperoxidase method to demonstrate intercellular immunoglobulins in the epidermis

In 3 dogs with pemphigus vulgaris and 4 dogs with pemphigus foliaceus, intercellular immunoglobulins were demonstrated in the epidermal stratum spinosum. The immunofluorescence technique on cold ethanol-fixed and paraffin-embedded tissue sections was compared with the immunoperoxidase method on formalin-fixed paraffin-embedded tissue sections. The results of both methods were identical. However, the advantage of the unlabeled antibody-enzyme method was that the same formalin-fixed tissue specimens could be used for conventional light microscopy, as well as for immunohistologic studies.     

Cyclosporin: applications in small animal dermatology

Abstract Cyclosporin has been increasingly used for the treatment of skin diseases in small animals. Reported uses include the treatment of atopy, cutaneous lupus erythematosus, feline acquired alopecia resembling pseudopelade of humans, pemphigus erythematosus, pemphigus foliaceus, perianal fistulae and sebaceous adenitis. In addition, cyclosporin has been used anecdotally for several other skin diseases. Few side effects have been noted at doses therapeutic for dermatologic diseases. Current suggestions for monitoring, and the value of trough cyclosporin serum concentrations for prediction of toxicity and efficacy are discussed.     

Pemphigus foliaceus-like drug reaction in a 3-month-old crossbreed dog treated for juvenile cellulitis

Two weeks after administration of routine vaccinations, a 3-month-old, male crossbreed dog developed cutaneous lesions that were clinically and histopathologically consistent with a diagnosis of juvenile cellulitis. The patient was treated with systemic antibacterial, antibacterial ophthalmic ointment and nonsteroidal anti-inflammatory drugs. Within 1 week, new skin lesions that were clinically, histopathologically and immunohistochemically consistent with pemphigus foliaceus developed. The latter completely resolved following drug withdrawal, suggesting a pemphigus foliaceus-like drug reaction. However, the initial lesions persisted and required glucocorticoid therapy to effect resolution. To the authors' knowledge, this is the first reported case of a puppy developing juvenile cellulitis followed by a pemphigus foliaceus-like reaction shortly thereafter, in response to drug therapy.     

Pemphigus foliaceus in two Shetland sheepdog littermates

Two female Shetland Sheepdog littermates simultaneously developed pemphigus foliaceus at 6 months of age. Three other littermates were not affected. One bitch (tricolored) was not treated, and the disease has remained active for 2 years. The other bitch (blue merle) has been successfully managed with glucocorticoids and gold salts.