IgE-positive epidermal Langerhans cells in allergic contact dermatitis lesions provoked in patients with atopic dermatitis

Summary To see whether or not IgE-bearing epidermal Langerhans cells are specific to skin lesions of atopic dermatitis (AD), we performed immunohistochemical and immunoelectron microscopic examinations of dinitrochlorobenzene (DNCB) contact dermatitis lesions provoked in uninvolved skin of eight patients with AD. In all of the eight examined, IgE-positive epidermal Langerhans cells were observed in the DNCB dermatitis lesions. Typical staining of anti-IgE was absent in the epidermis of normal-appearing skin of five patients with AD. Thus, it is likely that IgE positive epidermal Langerhans cells non-specifically occur in different eczematous diseases provoked in patients with AD.     

Immunophenotyping of the cutaneous infiltrate and of the mononuclear cells in the peripheral blood in patients with atopic dermatitis

Fourteen adult patients with chronic atopic dermatitis and active skin lesions had a skin biopsy and venous blood sample taken on the same day. Absolute numbers of circulating lymphocytes were normal in all patients. Fluorescence-activated cell sorter (FACS) analysis revealed normal numbers of total T lymphocytes and T-helper and T-suppressor subsets (helper:suppressor ratio, 2:1) in the atopic patients' peripheral blood, but an increase in circulating B lymphocytes and in HLA-D-related antigen-bearing cells. The skin biopsy showed a dermal infiltrate of predominantly T-helper lymphocytes (helper:suppressor ratio, 7:1). These cells showed strong HLA-DR plasma membrane staining. There was no HLA-DR staining in the membranes of epidermal keratinocytes. Using a monoclonal antihuman IgE, positive staining was observed in the dermis, though none was identified in the epidermis. The dermal anti-IgE staining was concentrated around clusters of T lymphocytes.     

Atopic dermatitis: immunophenotyping of inflammatory cells in skin lesions

BACKGROUND: The ever increasing incidence of atopic dermatitis (AD) has stimulated many researchers to use various diagnostic procedures to obtain new data to help elucidate the pathogenesis of the disease. AIMS: To perform cell immunophenotyping and to analyze the presence of inflammatory cell-surface markers in the biopsies of skin lesions from 15 AD patients and five healthy subjects. METHODS: Immunohistochemical analysis was performed in a group of AD patients and compared with that in a control group of healthy subjects. Avidin-biotin immunoperoxidase staining of paraffin-embedded, 4 microm skin sections, with semiquantitative counting of cells labeled with anti-CD3, anti-CD8, anti-CD20, anti-HLA-DR (HLA, human leukocyte antigen), and anti-immunoglobulin E (anti-IgE) primary antibodies, was used. RESULTS: The results of AD skin analysis showed a greater infiltration of CD3+ lymphocytes, especially of CD4+ subtype, compared with CD8+ lymphocytes. AD skin biopsy specimens also showed a higher intraepidermal HLA-DR+ Langerhans' cell count, the presence of HLA-DR on lymphocytes in the dermis, and higher intraepidermal expression of IgE+ cells compared with healthy controls. CONCLUSIONS: A statistically significant difference (P < 0.05) was found between the two groups for intradermal and intraepidermal CD3, CD4, and HLA-DR, intradermal CD8, and intraepidermal IgE+ cells. Immunophenotyping was found to be a useful diagnostic method in AD patients.     

Ultrastructural immunogold labelling of human langerhans cells enriched epidermal cell suspension

Summary Colloidal gold particles are well suited as markers in electron microscopy. Indirect immunogold staining was used to identify cell membrane antigens defined by monoclonal antibodies OKT6 and BL6 on human Langerhans cells (LC) in suspensions. Isolated epidermal cells were obtained by skin trypsinization and enriched or depleted in OKT6 positive on BL6 positive LC using the panning method: incubation of OKT6 or BL6 preincubated cells on immunoglobulin coated dishes. Indirect immunogold staining was then performed after prefixation in 2% paraformaldehyde. In LC enriched suspensions, only LC exhibited a specific membrane labelling with OKT6 or BL6 recognized by the presence of small evently distributed gold granules. Neither Birbeck granules, nor other cytoplasmic organelles, were labelled. No other epidermal cells were found positive. In LC depleted suspensions, no labelling was observed. Immunogold labelling on LC enriched suspensions after panning is now in progress for the qualitative evaluation and the quantitative analysis of cell surface constituants and antigens expressed by human dendritic epidermal cells.Presented at the Society for Investigative Dermatology and the European Society for Dermatological Research (Joint International Meeting Washington, April 27, May 1, 1983)     

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.     

Immunohistologic characterization of stasis dermatitis transformed skin of the lower leg

Antibodies against human T-lymphocytes and their subpopulations were applied to frozen sections obtained from skin affected with stasis dermatitis of the lower leg from 13 patients suffering from chronic venous insufficiency. In all cases, we observed an intense staining reaction with HLA-DR antibodies, i.e. mainly monocytes, macrophages, fibroblasts, and endothelial cells. Control sections of clinically normal skin taken from patients without stasis dermatitis showed only staining of endothelial cells and of dendritic epidermal cells with HLA-DR antigens. The dermal infiltrate in stasis dermatitis displayed only moderate staining with antigens directed against T-helper and T-suppressor cells. Our findings of large quantities of HLA-DR positive cells in skin affected with stasis dermatitis points to their possible role with regard to the induction and persistence of contact allergies so frequently encountered in this group of patients.     

Expression of the high affinity IgE-receptor on human Langerhans' cells. Elucidating the role of epidermal IgE in atopic eczema.

Epidermal Langerhans' cells have previously been shown to bear IgE molecules, particularly in atopic dermatitis skin. Using two highly specific antibodies against the antibody-binding chain of the high affinity IgE-receptor, 29C6 and 6F7, we here provide evidence that Langerhans' cells express this receptor in both normal skin (foreskin) and in lesional skin of patients with atopic and stasis eczema. A specific antibody against the low affinity IgE-receptor, Tü1, showed only a low expression of this receptor. This finding has important potential functional implications for the role of Langerhans' cells in transepidermal, IgE-mediated allergy.     

Occupational dermatitis in bakers: a clue for atopic contact dermatitis

6 patients are described who developed contact dermatitis after cereal contact on atopic skin for periods of 2 to 20 years. 2 patients were wheat flour patch-test-positive. They had punch biopsies taken for standard histological and immunohistochemical investigation by labeling with monoclonal antibodies, anti-DR and anti-IgE. Sections showed features of contact dermatitis. There were many dendritic cells located perivascularly in the papilla and in the epidermidis, intensely positive for monoclonal anti-IgE antibody. In control atopic subjects, there were a few perivascular IgE positive cells, probably mastocytes. This study shows that there may be a relationship between some allergens and atopic eczema in patients exposed to them in the course of their work. In some cases, there was a true allergic contact dermatitis, seen through the clinical and histological characteristics, and the results of immunohistochemical study.     

Mast cells and IgE in intestinal mucosa in adult atopic dermatitis patients

Duodenal biopsies from 29 adult atopic dermatitis (AD) patients with multiple positive skin prick test reactions were examined and the results compared with biopsies from 13 non-atopic controls. The duodenal mucosa showed mild inflammatory changes in six out of the 29 patients, but was normal in all the controls. Numerous anti-IgE positive cells, increasing with the severity of AD, were found in the duodenal mucosa in 25 of the 29 AD patients compared with few sporadic positive cells seen in only two out of 13 controls (P less than 0.001). The total serum IgE level showed a significant positive correlation with the number of anti-IgE stained cells in the mucosa (P less than 0.05). No significant differences were found in the total number of toluidine blue stained cells or cells immunoreactive for histamine between patients and controls. However, AD patients who had high numbers of anti-IgE positive cells often had decreased numbers of histamine immunoreactive cells in the mucosa suggesting mast cell degranulation. These findings provide further evidence that also in adult AD patients the gastrointestinal tract may serve as a portal of entry for allergens which may lead to exacerbation of AD.     

Immediate and delayed hypersensitivity reactions to birch pollen in patients with atopic dermatitis.

We investigated immediate and delayed hypersensitivity to birch pollen in 10 patients with atopic dermatitis (AD) who had experienced a worsening of their eczema during the birch pollen season. The patients were prick- and patch-tested and antigen-induced basophil histamine release and lymphocyte proliferation were measured. 9/10 birch pollen-allergic patients proved positive in the histamine release test and the results correlated with specific IgE levels measured by RAST. Birch pollen antigen induced lymphocyte proliferation in 6/10 patients, but a positive patch test result was obtained in only one case. Both peripheral blood monocytes and purified epidermal Langerhans' cells were able to present birch pollen antigen to T cells, although Langerhans' s cells seemed to function less efficiently in this respect.     

Plasmacytoid dendritic cells: a new cutaneous dendritic cell subset with distinct role in inflammatory skin diseases

Epidermal dendritic cells found in inflamed skin include Langerhans cells and the recently identified population of inflammatory dendritic epidermal cells. Another subset of dendritic cells in humans is the plasmacytoid dendritic cell in peripheral blood, which is characterized by the production of large amounts of type I interferon (interferon-alpha and interferon-beta) upon viral infection. We hypothesized that plasmacytoid dendritic cells might be involved in anti-viral defense mechanisms of the skin. Here we investigated plasmacytoid dendritic cells, inflammatory dendritic epidermal cells, and Langerhans cells in epidermal single cell suspensions of normal looking skin from healthy volunteers and of lesional skin from patients with different inflammatory skin diseases. Langerhans cells were found in normal and in inflamed skin samples. In normal skin, plasmacytoid dendritic cells and inflammatory dendritic epidermal cells were low or absent. Lesional skin samples from patients with psoriasis vulgaris and contact dermatitis contained relatively high numbers of both inflammatory dendritic epidermal cells and plasmacytoid dendritic cells. In contrast, many inflammatory dendritic epidermal cells but only very few plasmacytoid dendritic cells could be detected in atopic dermatitis lesions. Lupus erythematosus was characterized by high numbers of plasmacytoid dendritic cells but low numbers of inflammatory dendritic epidermal cells. These results demonstrate that in addition to resident Langerhans cells, plasmacytoid dendritic cells and inflammatory dendritic epidermal cells are selectively recruited to the skin lesions depending on the type of skin disease. The lack of plasmacytoid dendritic cells in atopic dermatitis may predispose atopic dermatitis patients to viral infections such as eczema herpeticum, a secondary infection of atopic dermatitis lesions with herpes simplex virus. The composition of dendritic cell subsets may help to clarify the etiology of inflammatory skin diseases and forms the basis for therapeutic intervention with selective microbial molecules such as immunostimulatory CpG oligonucleotides.     

The cell population in the cutaneous infiltrate of mycosis fungoides: in situ studies using monoclonal antisera

T cell antigens were studied in cutaneous sections from five patients with mycosis fungoides (MF). The method allowed cell counting to be undertaken for each monoclonal antiserum. OKT3 (pan T cell) antiserum confirmed the predominantly T lymphocytic nature of the infiltrate, labelling the majority of infiltrating cells. OKT4 (helper/inducer) antiserum positively labelled 90% of the lymphocytes identified as OKT3⁺. OKT8 (suppressor) antiserum marked only single or small groups of dermal lymphocytes, which comprised 24% of the cells identified as T lymphocytes. OKT6 (anti-Langerhans) showed positive labelling of dendritic cells in the epidermis and dermis. Fewer positively labelled epidermal dendritic cells were observed in sections from patients receiving PUVA, but no difference was found in the number of OKT6 positive dermal cells. The ratio of helper to suppressor cells in the dermal infiltrate significantly exceeded the normal circulating ratio.     

Abundant immunoglobulin E‐positive cells in skin lesions support an allergic etiology of atopic dermatitis in the elderly

Background/Objectives Atopic dermatitis (AD) in the elderly is gradually increasing in industrialized countries in association with the aging of society. We report herein four cases of elderly AD {three extrinsic [immunoglobulin (Ig)E‐mediated allergy]; one intrinsic (non‐IgE‐allergy)} in which we investigated the presence of IgE+ cells in lesional skin. Methods/Results Single immunohistochemical and double immunofluorescence stainings were performed for skin biopsy specimens from AD patients and non‐atopic control subjects with chronic eczema. In the lesional lichenified skin of patients with extrinsic elderly AD, numerous IgE+ cells were found among inflammatory cells infiltrates in the upper dermis. Comparative analysis of single immunohistochemistry results using serial paraffin and/or frozen sections found that many IgE+ cells showed identical distributions to tryptase+ mast cells. IgE+ cells coincident with CD1a+ Langerhans cells in the epidermis were found in small numbers only in frozen sections. Double immunofluorescence staining for IgE and CD11c revealed cells coexpressing IgE and CD11c with a dendritic morphology in the papillary and upper dermis. These IgE+ mast cells and IgE+ CD11c+ cells were also found in cured normal‐looking skin from a patient with extrinsic elderly AD after successful treatment. Although only a few weakly positive IgE+ cells were detected, no IgE+CD11c+ cells were found in specimens from patients with intrinsic elderly AD or non‐atopic chronic eczema. Conclusion IgE‐mediated allergic inflammation may play an important role in the pathobiology of elderly AD, similar to other age groups of AD.     

Evaluation of Langerhans' cells in normal and eczematous dermatitis skin by CD1a protein immunohistochemistry: preliminary findings

Background:  Langerhans' cells (CD1a positive, bone marrow–derived cells), are the antigen presenting cells of the skin. Our knowledge about the status of these cells in eczematous dermatitis is incomplete.
Aim:  This study tests the hypothesis that 'the development of eczematous dermatitis is associated with alterations of Langerhans' cells'.
Materials and methods:  Biopsy specimens from patients with eczematous dermatitis and normal skin (20 cases, each) were studied. Langerhans' cells were stained for CD1a using imunoperoxidase-staining methods and mouse monoclonal antibodies.
Results:  In normal skin, CD1a+ Langerhans' cells were seen in suprabasal position. In eczematous dermatitis skin, CD1a positive cells were seen scattered in the acanthotic epidermis. Compared with normal skin, the mean values of the Langerhans' cells were statistically significantly higher in eczematous dermatitis [epidermal Langerhans' cells: 1.20 (standard error of mean, SEM, 0.13) vs. 2.50 (SEM, 0.16); and dermal Langerhans' cells: 1.30 (SEM, 0.15) vs. 2.7 (SEM, 0.15); for normal and eczematous skin, respectively; p < 0.05].
Conclusions:  The higher Langerhans' cell counts in eczematous dermatitis suggest a possible link between antigen presenting capabilities of these cells, and development of these lesions.     

Human epidermal Langerhans' cells in bullous pemphigoid

Through the epidermal analysis of 13 patients with bullous pemphigoid compared to controls, using OKT6 monoclonal antibodies on the light microscopic level and electron-microscopy, we found a redistribution of the Langerhans' cells towards the basal membrane in combination with an increased total number of Langerhans' cells. This redistribution was also noted in clinically normal skin from patients with bullous pemphigoid. The findings may be consistent with the theory of antigen presentation.     

An immunohistochemical staining of epidermal Langerhans' cells in tinea cruris

Epidermal Langerhans' cells (LC) were investigated in fresh cryostat sections of ten biopsies from patients with mycologically proven tinea cruris, using OKT6 monoclonal antibodies and avidin-biotin-immunoperoxidase. Compared to the controls, more epidermal LC and an increased number of LC in the upper half of the epidermis were found in the sections from tinea patients. In a double staining method for both OKT6-positivity and hyphae, a tendency towards a gathering of LC and fungal elements was found. The results of this study are in agreement with the theory that epidermal LC are responsible for the antigen uptake in dermatophytosis.     

Immunoglobulins and Their Receptors on Epidermal Langerhans Cells in Atopic Dermatitis

To elucidate the etiological role of immunoglobulin molecules on Langerhans cells (LCs) in atopic dermatitis, we conducted immunohistochemical studies on the localization of immunoglobulin G1 (IgG1), IgG2, IgG3, IgG4, IgA and IgM on epidermal LCs from 30 patients with atopic dermatitis (AD) and five non-atopic healthy volunteers. We also investigated the types of receptors for the immunoglobulins (FcεRI, FcεRII, FcγRI, FcγRII, and FcγRIII) on epidermal LCs in the patients. IgE positive epidermal LCs were observed in 28 of 30 AD patients, and 46.7% of the epidermal LCs were positive for IgE. Both IgG1- and IgG2-positive epidermal LCs were obserbed in 70% of AD patients, and 21.8% and 28.7% of the total epidermal LCs were positive for IgG1 and IgG2, respectively. IgG3- or IgG4-positive LCs were present in only small proportions of AD patients. IgA-positive LCs were observed in 8 AD patients; our study suggested that the IgA bound on LCs was secretory IgA (S-IgA). These surface immunoglobulins were observed significantly more frequently on epidermal LCs in the involved skin of AD than in clinically uninvolved skin. No IgM-positive epidermal LCs were observed in the AD patients or healthy volunteers. In non-atopic healthy controls, no immunoglobulin-binding LCs were observed. In receptors for immunoglobulins, FcεRI and FcγRII were exclusively expressed on nearly all epidermal LCs from all AD patients and all non-atopic controls. These results suggested that not only IgE but also IgG and IgA may play some etiological role in the pathogenesis of AD.     

LEKTI demonstrable by immunohistochemistry of the skin: a potential diagnostic skin test for Netherton syndrome

BACKGROUND: Netherton syndrome (NS) is a rare autosomal recessive condition characterized by ichthyosiform erythroderma, trichorrhexis invaginata and atopic manifestations. Confirming the diagnosis may be difficult in the early stages. Mutations in the SPINK5 gene which encodes for the serine protease inhibitor LEKTI are associated with NS. These mutations create premature termination codons which result in absent or abnormal expression of LEKTI in patients with NS. OBJECTIVES: To investigate the expression of LEKTI in the skin of patients with NS in comparison with normal controls and patients with other skin conditions, namely atopic dermatitis, psoriasis and nonbullous ichthyosiform erythroderma. METHODS: Immunohistochemistry was performed on skin sections from four patients with NS, four normal controls, four with atopic dermatitis, two with psoriasis and two with nonbullous ichthyosiform erythroderma, using a primary rabbit polyclonal antibody against LEKTI. RESULTS: LEKTI was localized to the stratum granulosum in normal skin. All four skin sections from patients with NS showed absent or very reduced staining for LEKTI. Staining in the other disorders showed positive LEKTI expression in varying patterns. CONCLUSIONS: NS can be difficult to diagnose especially in the early stage, which can lead to inappropriate treatments particularly if it is misdiagnosed as atopic dermatitis. Immunohistochemistry of skin with an antibody against LEKTI is a potentially useful diagnostic test for NS.     

Dorfman-Chanarin syndrome in a Turkish kindred: conductor diagnosis requires analysis of multiple eosinophils

Dorfman-Chanarin syndrome is a rare, autosomal recessive inherited lipid storage disease with congenital ichthyotic erythroderma due to an acylglycerol recycling defect. Demonstration of lipid vacuoles in neutrophils from peripheral blood smears (Jordans' anomaly) in patients with ichthyotic erythroderma leads to the diagnosis. In spite of frequent liver, muscle, ear, eye and central nervous system involvement, Dorfman-Chanarin syndrome may present clinically as monosymptomatic ichthyosis. Here, we report clinical and laboratory investigations in a consanguineous family from Turkey with 3 affected family members, and demonstrate the lipid vacuoles in epidermal Langerhans' cells for the first time. Langerhans' cell phenotyping suggests that the skin inflammation is due to the gene defect and not to underlying atopic dermatitis. Microscopic examination of eosinophils for lipid vacuoles to identify conductors revealed variable percentages of normal and vacuolized eosinophils in conductors, suggesting the microscopic analysis of at least 10 eosinophils for conductor identification.     

Occurrence of IgE-bearing epidermal Langerhans cells in atopic eczema: a study of the time course of the lesions and with regard to the IgE serum level

Uninvolved and lesional skin of untreated and treated patients with atopic eczema has been investigated immunohistochemically to determine the conditions in which IgE-bearing CD1a+ Langerhans cells/indeterminate cells (LC/IC) occur in this disease. IgE-bearing epidermal dendritic cells were present in patients with elevated IgE serum level (greater than 300 UI/ml) and the staining pattern was stronger in lesional skin. On double immunostaining, a subpopulation of CD1a+ LC/IC was found not to bear IgE molecules as determined by the ratio IgE+/CD1a+ cells on serial sections as well. The ratio IgE+/CD1a+ cells decreased in patients who underwent a local therapy with glucocorticosteroids. These results suggest that the expression of IgE receptors and/or binding of IgE molecules on epidermal LC/IC in atopic eczema may be controlled by a complex network of mediators from the epidermis or the inflammatory infiltrate, or both, and that this phenomenon could be down regulated by glucocorticosteroids.