Direct Immunofluorescence Of Perilesional Skin Research Articles

DP17 A very rare presentation of linear IgA disease during pregnancy

Abstract We present a very rare immunobullous dermatosis, linear IgA disease (LAD), that developed during pregnancy, to highlight its clinical features, management and important safety considerations for the neonate. During her first pregnancy, a 32-year-old Japanese woman of 27 weeks’ gestation developed an intensely itchy blistering rash on her limbs, trunk and face without periumbilical predilection. The blisters were tense, clustering in an annular configuration, with subsequent development of blistering and erosions involving the oral and genital mucosae. Medications included a pregnancy multivitamin only. A lesional skin biopsy revealed a subepidermal blister containing neutrophils and eosinophils. Direct immunofluorescence of perilesional skin demonstrated linear deposition of IgA along the basement membrane zone. Linear IgA disease was diagnosed. Initial treatment consisted of erythromycin and daily clobetasol ointment. Owing to inadequate symptom control and blister progression, dapsone 50 mg daily was initiated with prompt improvement in her condition and cessation of new blistering. A postpartum flare necessitated a dose increase to 75 mg daily. Her baby’s glucose-6-phosphate dehydrogenase (G6PD) level was found to be normal at birth. She proceeded to breastfeed her son, and monitoring of his haemoglobin level was normal. Although her clinical presentation initially raised the possibility of pemphigoid gestationis (PG), there was no periumbilical involvement, and the morphology of blistering in conjunction with mucosal involvement was more suggestive of LAD. The immunofluorescence results also helped to distinguish LAD from PG. The new onset of LAD during pregnancy is very unusual and has been rarely reported (Ikkaku N, Tateishi N, Oda Y et al. Linear immunoglobulin A bullous dermatosis developing during late pregnancy. J Dermatol 2017; 44:e44–5). Pre-existing LAD typically improves during pregnancy, although postpartum flares are recognized. Pemphigoid gestationis has been associated with an increased risk of prematurity and intrauterine growth restriction, but this has not been reported with LAD during pregnancy, possibly due to its rarity. Both the diagnosis of LAD and its mainstay treatment—dapsone—have important potential implications for the neonate. As for other immunobullous diseases, transplacental transfer of pathogenic antibodies may result in transient, usually mild, neonatal blistering. Although dapsone can be safely used during pregnancy and breastfeeding, the risk of neonatal haemolysis and methemoglobinaemia necessitates the assessment of the G6PD level and haemoglobin monitoring in the neonate. Multidisciplinary input from the dermatology, obstetric and paediatric teams is crucial to ensure appropriate counselling and care of the affected mother and baby.

A case report pemphigoid gestational case, responding to systemic steroid therapy

In this case report, a rare case of Pemphigoid Gestationis at 33 weeks’ of gestation is presented and the related literature is briefly reviewed. A 21-year-old at 33 weeks’ of gestation admitted with the complain of itchy skin lesions on the abdomen, arms and legs. Dermatologic examination revealed urticaria papules and plaques in the abdomen, arms and legs, more intensely around the umbilicus, which range from 1-2 cm diameter to 4-5 cm diameter. Papulovesicular lesions and few erosions were observed especially in the hand and foot dorsals. Spongiosis on epidermis, intracellular and subepidermal bullae formation, edema on papillary architecture and inflammatory infiltrate composed of mononuclear cells and eosinophils around the perivascular ares of the upper dermis were seen in histological examination. Direct immunofluorescence of perilesional skin showed linear deposition of complement (C3) along the basement membrane zone. The preliminary diagnosis of pemphigoid gestationis was confirmed by histopathologic findings. Pemphigoid gestationis is a rare autoimmune bullous dermatosis that begins in the second or third trimester of pregnancy. Healing occurs weeks after birth. It occurs more severely in subsequent pregnancies. Its main treatment is systemic steroid and antihistamine administration. A multidisciplinary approach both in dermatological and gynecological terms is required for the diagnosis, treatment and course of the disease.

Pemphigus vulgaris as the first manifestation of multiple myeloma: a case report

BackgroundThe association between pemphigus and malignancy has been well documented for decades but an association between pemphigus vulgaris and multiple myeloma is unusual. We report a case of pemphigus vulgaris revealing multiple myeloma.Case presentationA 55-year-old Malagasy man, with no significant past medical history, presented with bullous and erosive skin lesions involving his trunk and scalp for the last 2 months. He had no mucous membrane involvement. A diagnosis of pemphigus vulgaris was made on skin biopsy and direct immunofluorescence of perilesional skin revealing immunoglobulin G deposition in the intercellular spaces in the epidermis. In an enzyme-linked immunosorbent assay, his serum autoantibody index against desmoglein-1 and 3 was found to be 112 RU/mL and 34 RU/mL respectively. Serum immunoelectrophoresis showed a monoclonal gammopathy with a markedly elevated immunoglobulin G level (2880 mg/dL) in association with a lambda free light chain. Bone marrow aspirate showed 6% plasma cell infiltration. Further investigations, including creatinine blood test and whole body radiographic examinations, showed that he had initially clinical stage I multiple myeloma of the immunoglobulin G-λ type. Six months later, bone tomography revealed vertebral compression fractures of the thoracic and lumbar spine that correlated with his back pain topographically. Anti-myeloma treatment including melphalan and prednisone led to an immediate decline in monoclonal immunoglobulin G concentration. Skin and hematologic remission were maintained for 12 months.ConclusionsAbsence of mucosal involvement, lack of vacuolar degeneration at the interface, and absence of apoptotic, dyskeratotic keratinocytes ruled out paraneoplastic pemphigus in our case. Pemphigus vulgaris should be considered even if possible underlying disease for which paraneoplastic pemphigus is recognized is present.

To compare the sensitivity pattern of direct immunofluorescence of perilesional skin and plucked hair in pemphigus vulgaris

To compare the sensitivity pattern of direct immunofluorescence of perilesional skin and plucked hair in pemphigus vulgaris

Three case reports of post immunization and post viral Bullous Pemphigoid: looking for the right trigger

BackgroundBullous pemphigoid (BP) is a blistering skin disorder infrequent in infancy and rarely reported in medical literature.Case PresentationHere we describe three cases of BP which were referred to our department in the last 15 years. Two of them developed an eruption of bullous lesions just a few days after vaccination for diphtheria, tetanus, pertussis, poliomyelitis, hepatitis B and Haemophilus influenzae B. The third patient developed the same blistering lesions shortly after herpetic stomatitis. In all three cases, clinical diagnosis was confirmed by histological examination which showed subepidermal bullae with a dermal inflammatory infiltrate, and direct immunofluorescence of perilesional skin showed linear IgG and C3 deposits along the basement membrane zone. Immunoblot assay was positive for BP antigen 180. Treatment with oral prednisone was instituted and the lesions resolved in two out of three patients; the third one was treated with an immunosuppressive agent (tacrolimus) and corticosteroid and subsequently with intravenous immunoglobulin and plasmapheresis, due to an underlying complex autoimmune disease.ConclusionAlthough the mechanism of induction of BP is still unclear, the close relationship between trigger events (immunization or viral infection) and onset of the disease arises a possible association.

IgA LINEAR BULLOUS DERMATOSIS IN CHILDHOOD

IgA linear bullous dermatosis, also known as chronic bullous dermatosis of childhood, is an autoimmune disease which may be idiopathic or drug-induced. The disease affects children and adults. We present a 4 years old girl with itchy polymorphic eruptions. The skin rash was presented by bullous-erosive rosette-like lesions with reddish-brown crust in the center, distributed on the skin of the face, trunk and extremities. The vesicles were filled with serous and hemorrhagic content. Laboratory examinations were within normal values according the age. Histopathological examination of the lesional skin revealed sub epidermal blister. Direct immunofluorescence of perilesional skin demonstrated linear deposition of IgA in the basement membrane. Systemic treatment with Methylprednisolon and Claritromycin was applied with satisfactory effect. The patient is under observation.

DERMATITE HERPETIFORME EM IDADE PEDIÁTRICA – UM DIAGNÓSTICO A TER EM CONTA

Introdução: A Dermatite herpetiforme é uma dermatose crónica, pruriginosa e polimórfica considerada o equivalente cutâneo da doença celíaca. Na população pediátrica é, com frequência, um diagnóstico difícil de evocar, quer pela sua raridade quer pelo polimorfismo das lesões cutâneas.Caso Clínico: Apresentamos um doente do sexo masculino de 8 anos, com lesões polimorfas nas superfícies extensoras dos membros evoluindo há 2 anos. A biopsia cutânea lesional demonstrou neutrófilos nas papilas dérmicas e a imunofluorescência directa peri-lesional revelou depósitos granulosos de IgA nas papilas dérmicas, estabelecendo o diagnóstico de dermatite herpetiforme. Estudo complementar subsequente confirmou a associação a doença celíaca e a adopção de uma dieta sem glúten levou à resolução clínica cutânea e da enteropatia.

IgG/IgA pemphigus reactive with desmoglein 1 with additional undetermined reactivity with epidermal basement membrane zone

IgG/IgA pemphigus is an extremely rare subset of pemphigus, showing anti-keratinocyte cell surface antibodies of both IgG and IgA classes. Herein, we describe a unique case of IgG/IgA pemphigus with clinical features of edematous erythema and peripheral vesiculopustules. Histopathology showed the presence of subcorneal pustules and acantholytic blisters in the mid-epidermis with neutrophilic infiltration and eosinophilic spongiosis. Direct immunofluorescence of perilesional skin showed both IgG and IgA deposits to keratinocyte cell surfaces and unusual granular deposits of IgG, IgM, and C3 along basement membrane zone. On enzyme linked immunosorbent assay , the auto-antibodies were found to be reactive to desmoglein 1 antigen. Various clinical, histopathological, and immunological findings in our case overlapped with the features of IgA pemphigus, pemphigus herpetiformis, and pemphigus foliaceus. These findings indicate that IgG/IgA pemphigus may be a transitional form between IgA pemphigus and pemphigus herpetiformis, and thus provides insight into the pathogenicity of this rare disorder.

Direct immunofluorescence of the outer root sheath in anagen and telogen hair in pemphigus vulgaris and pemphigus foliaceus

Direct immunofluorescence of peri-lesional skin is the gold standard in the diagnosis of pemphigus. A specific immunofluorescence pattern may also be demonstrated in the outer root sheath of anagen and telogen hair. We demonstrated an intercellular reticular deposition of immunoglobulin G in the outer root sheath of plucked anagen and telogen hair in all pemphigus vulgaris patients with active disease and for the first time in all patients with active pemphigus foliaceus. Moreover, we demonstrated for the first time that plucked hair samples may be kept at -20°C for at least 2 weeks before immunofluorescent staining and analysis.

Juvenile Bullous Pemphigoid – a Case Report

Abstract Bullous pemphigoid is an autoimmune blistering disease that predominantly affects elderly persons and rarely children. We present a 12-year-old girl with sudden appearance of tense blisters on an erythematous base on the trunk, neck, hands and legs with intense pruritus. Standard laboratory test results were within the normal range except for blood eosinophilia of 12% of the total white cell count. Skin biopsy specimens showed evolving subepidermal blisters with perivascular lymphohistiocytic, eosinophil and neutrophil infiltrations in the papillary dermis. Direct immunofluorescence of perilesional skin showed linear, continuous deposits of IgG and C3 along the dermoepidermal junction. Indirect immunofluorescence showed circulating anti-basement membrane zone IgG autoantibodies at a titer of 1:80. We started treatment with systemic corticosteroids, methylprednisolone 0,5 mg/kg per day and 500 mg erythromycin 4 times a day during 10 days. After 3 days 50 mg dapsone (DDS, 4,4-diaminodiphenylsulphone) per day was added. After a few days, there were no new changes on the skin and pruritus disappeared completely.

Linear IgA bullous dermatosis

Linear IgA disease is a rare chronic autoimmune subepidermal blistering disorder characterized by the linear deposition of IgA along the dermoepidermal junction. A 40-year-old housewife presented with symmetrically distributed generalized intensely pruritic vesiculobullous lesions of 1 year duration. Previous treatment with oral and topical steroids had induced only a temporary remission of the skin lesions. There was no history of drug intake in the recent past and her medical history was insignificant and there was no similar illness in her family members. Examination findings included vesicles arranged at the periphery of healed lesions along with post-inflammatory hyperpigmented macules on the trunk and extremities. There was no genital, mucosal or ocular involvement. Skin biopsy from the leg showed dense neutrophilic infiltration along with eosinophils at the tip of the dermal papillae and focal areas showed dermoepidermal clefting with bulla formation. Direct immunofluorescence of perilesional skin revealed linear deposits of IgA at the basement membrane zone. Therapeutically the patient responded well to dapsone. Though the presentation was classical, the case is reported for its rarity.

Bullöse Autoimmundermatosen

Autoimmune bullous skin disorders are rare, potentially fatal disorders of skin and mucous membranes which are associated with IgG or IgA autoantibodies against distinct adhesion molecules of the epidermis and dermal epidermal basement membrane zone, respectively. These autoantibodies lead to a loss of skin adhesion which shows up clinically as the formation of blisters or erosions. In pemphigus, loss of adhesion occurs within the epidermis while in the pemphigoids, linear IgA dermatosis, epidermolysis bullosa acquisita and dermatitis herpetiformis, loss of adhesion takes place within or underneath the basement membrane zone. The autoantigens of these disorders are largely identified and characterized. Making the diagnosis of autoimmune bullous skin diseases is based on histology and direct immunofluorescence of perilesional skin and the serological detection of autoantibodides by indirect immunofluorescence and recombinant autoantigens. Therapeutically, systemic treatment with glucocorticoids is combined with immunosuppressive adjuvants which allow for the fast reduction of systemic steroids. A prospective trial in pemphigus showed that adjuvant treatment with azathioprine, mycophenolate mofetil and cyclophosphamide, respectively, led to a significant reduction of the cummulative dose of systemic steroids until complete clinical remission was achieved. In bullous pemphigoid, topical treatment with clobetasol led to complete clinical remissions without major side effects seen when glucocorticoids were applied systemically. Therapeutic depletion of B cells by rituximab as a second line therapy has significantly improved the overall prognosis of pemphigus. Comparable controlled therapeutic trials have not yet been performed in dermatitis herpetiformis and epidermolysis bullosa acquisita.

A Current Review of Juvenile Pemphigus Vulgaris

Forty-seven cases of juvenile pemphigus vulgaris have been reported in the English literature. Histology of lesional skin and direct immunofluorescence of perilesional skin are both necessary for a complete diagnosis. The autoimmune bullous condition can affect the skin and mucous membranes individually, but typically affects both concurrently. Disease characteristics in juvenile patients are similar to those in adults; however, a disruption of biologic and social development is of particular concern during adolescence. Although systemic corticosteroids have been used to successfully treat the disease in most cases, long-term use is often necessary for adequate control. Adverse effects from therapy can have devastating effects during this critical period of hormonal changes, physical and mental growth, and social and cultural development that occurs during adolescence. Newer therapies must be designed to adequately treat juvenile patients while also limiting serious adverse effects.

Two cases of palmar petechiae as a presentation of dermatitis herpetiformis

Conflict of interest: none declared. Haemorrhagic palmoplantar lesions are a recognised presentation of childhood dermatitis herpetiformis (DH), with a few reported adult cases.1, 2 We report two cases of palmar petechiae as a presenting feature of adult DH. Patient 1 was a 34‐year‐old woman, who presented with an 18‐year history of a rash affecting her fingertips and toes, most prominent on her dominant hand and preceded by an itchy sensation. On physical examination, multiple petechial lesions were seen on her fingertips and toes (Fig. 1a), together with a small tense blister filled with serosanguinous fluid on the right hand. Screening for anti‐mitochondrial, anti‐smooth muscle, anti‐parietal cell, anti‐liver kidney microsomal antibodies, cytoplasmic antinuclear cytoplasmic antibodies (ANCA) perinuclear ANCA, thrombophilia and porphyria was negative. She had a weakly positive antinuclear antibody titre with a nucleolar pattern. Histological examination of a petechial lesion found disruption of the keratin layer with haemorrhage, intracorneal vesicle formation and red cell extravasation in the dermis with moderate oedema. Histological examination of a biopsy taken from the blister on the right hand revealed a subepidermal split with abundant neutrophils, lymphocytes and histiocytes. Effacement of the dermal papillae was noted (Fig. 1b). Direct immunofluorescence of perilesional skin showed fibrillar deposits of IgA and fibrinogen within the dermal papillae (Fig. 1c). Anti‐endomysial IgA antibodies were negative on indirect immunofluorescence, but duodenal biopsies confirmed subtotal villous atrophy (Fig. 1d).

Demonstration of pemphigus‐specific immunofluorescence pattern by direct immunofluorescence of plucked hair

Direct immunofluorescence (DIF) of perilesional skin is the gold standard in the diagnosis of pemphigus. Since the outer root sheath (ORS) of anagen hair is structurally analogous to epidermal keratinocytes, pemphigus specific immunofluorescence pattern may be present in the ORS. Twenty consecutive patients of pemphigus were enrolled in the study, irrespective of any other inclusion or exclusion criteria. Hairs were plucked in a similar fashion to that of trichogram. Approximately 5 anagen hairs were selected processed and stained with fluorescein isothiocyanate (FITC) conjugates. DIF of perilesional skin and indirect immunofluorescence (IIF) were carried out simultaneously. DIF of hair was also done in equal numbers of controls with other dermatoses. Intercellular deposition of IgG was seen in the ORS of anagen hair in 85% of patients (n = 17). The test was positive in all patients who had scalp lesions (n = 10); however, it was also positive in 7 patients (77.7%) who did not have scalp lesions. The test was negative in 3 patients; two of whom were elderly and had sparse scalp hair. The other patient only had mucosal lesion. The test was negative in the control group. DIF of hair is a simple, non-invasive test. In future, it may alleviate the need for skin biopsies, in patients with pemphigus.

A case of linear IgA disease in a child with IgA and IgG circulating antibodies directed to BPAg2

A 7-year-old Caucasian girl had multiple bullae on the trunk, upper and lower limbs (Fig. 1a,b) for 10 days. The lesions were large, tense, and asymptomatic, and mucosae were not involved. Laboratory findings were all normal. Histopathology revealed a subepidermal blister containing numerous neutrophils, eosinophils, and fibrin. Direct immunofluorescence of perilesional skin disclosed linear deposition of IgA and slight linear deposits of IgM at the basement membrane zone (Fig. 2). Indirect immunofluorescence on monkey esophagus and on human salt-split skin was negative. Immunoblot assay (IB), performed with an epidermal extract, revealed IgA and IgG antibodies directed to BPAg2 antigen (Fig. 3), while it was negative when performed on dermal extract. Enzyme-linked immunosorbent serologic assay using a commercial kit (MBL, Naka-Ku Nagoya, Japan) with the noncollagenous domain (NC16A) of the BPAg2 antigen was negative for both IgA and IgG. A diagnosis of linear IgA disease (LAD) was made and a treatment with dapsone (50 mg/day) and prednisolone (30 mg/day) was initiated. One month later, lesions had cleared.

Autoimmune bullous disorders

Bullous skin diseases represent a group of organ-specific autoimmune disorders characterised by binding of circulating autoantibodies to adhesion molecules of the epidermis and the dermo-epidermal basement membrane zone. Binding of these autoantibodies to their antigenic targets results in loss of adhesion between epidermal keratinocytes and at the level of the basement membrane zone. Chronic blisters and secondary painful erosions are the clinical hallmark of autoimmune bullous disorders. Histopathology reveals the location of blister formation and helps to classify the subtype of the bullous skin disorder. Immunofluorescence is crucial for diagnosing autoimmune bullous skin disorders. Tissue-bound autoantibodies are detected by direct immunofluorescence of perilesional skin. Circulating autoantibodies can be visualised by indirect immunofluorescence using tissue substrates such as monkey oesophagus and sodium chloride-split human skin. Most of the autoantigens are available as recombinant proteins, which allows for autoantibody screening by ELISA or immunoblot analysis to confirm the primary diagnosis and, importantly, for immunoserological follow-up of patients.

Inflammatory epidermolysis bullosa acquisita with coexistent IgA antibodies to plectin

We present a case of inflammatory epidermolysis bullosa acquisita (EBA) with IgA antibodies to plectin. Analysis of lesional skin biopsies by electron microscopy revealed the split level to be in the sublamina densa zone, corresponding to the diagnosis of EBA. Direct immunofluorescence of perilesional skin demonstrated u-serrated depositions of IgG and IgA that under immunoelectron microscopy were shown to be located in the sublamina densa. In contrast, indirect immunofluorescence on salt-split skin revealed circulating IgA antibodies that stained the roof rather than the floor of the blister. Immunoblotting showed these serum antibodies to be directed to the cytoplasmic hemidesmosomal antigen plectin. The antiplectin specificity of these antibodies was confirmed by 'knockout' immunofluorescence analysis; the serum IgA did not bind to skin sections of a patient with plectin-deficient epidermolysis bullosa. To our knowledge, this case demonstrates for the first time the existence of IgA antibodies against plectin.

Linear IgA disease associated with lymphocytic colitis

A 66-year-old woman presented with a bullous skin eruption and chronic diarrhoea. Lesional skin showed subepidermal blistering, and direct immunofluorescence of perilesional skin revealed linear deposits of IgA at the dermoepidermal junction, establishing a diagnosis of linear IgA disease (LAD). Chronic watery diarrhoea complicated by substantial loss of body weight preceded the skin eruption for several months. On endoscopy, the colon appeared macroscopically normal. On histology, the colon mucosa showed increased numbers of intraepithelial lymphocytes and infiltrates of mononuclear cells in the lamina propria, indicative of lymphocytic colitis. Treatment with methylprednisolone and dapsone led to complete clearing of the bullous skin eruption and marked improvement of the patient's diarrhoea. Gastrointestinal disorders such as lymphocytic colitis have rarely been reported in patients with LAD. Whether the simultaneous occurrence of these two diseases is coincidental or due to related pathogenetic mechanisms remains to be seen.

Epidermolysis Bullosa Acquisita in Childhood

Epidermolysis bullosa acquisita (EBA), rarely reported in childhood, is described in a 10-year-old black girl. The age of onset during infancy and the clinical appearance mimicked hereditary dystrophic epidermolysis bullosa. Epidermolysis bullosa acquisita was diagnosed by direct immunofluorescence of perilesional skin, indirect immunofluorescence on normal epithelium and saline-split skin, direct immunoelectron microscopy, and immunoblotting of the patient's serum sample against partially purified EBA antigen/carboxyl domain of type VII collagen. Differentiation of mechanobullous disease in children is critical in that significant clinical benefit may be achieved in EBA with prednisone and/or dapsone therapy. A search for associated immunologic abnormalities and HLA-DR typing may help our understanding of EBA.