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.     

Subpopulations of T lymphocytes in peripheral blood and in skin lesions in lichen ruber planus

Peripheral blood lymphocytes and different immunocompetent cells of the cutaneous inflammatory infiltrate were characterized by reactivity with monoclonal antibodies directed to surface antigens of T lymphocytes and Langerhans cells by means of the indirect immunofluorescence technique in 10 patients suffering from lichen ruber planus. In contrast to the controls (n = 10), lichen patients had a significantly reduced percentage of the suppressor/cytotoxic T cell subset in peripheral blood (p less than 0.002). At the dermoepidermal junction suppressor/cytotoxic T lymphocytes and Langerhans cells predominated. Some possible conclusions are discussed.     

Predominant cutaneous infiltration by OKT6- and OKT8-positive cells in a case of Sézary syndrome

Summary Using an immunoperoxidase (skin biopsy) and an immunofluorescence (peripheral blood, bone marrow punctate) technique, and monoclonal antibodies raised against peripheral mature lymphocytes, T helper subsets, T suppressor subsets, and Langerhans cells, we found a predominant dermal infiltration with lymphocytes of the suppressor phenotype and a predominant epidermal infiltration with Langerhans cells in a patient with Sézary syndrome (cutaneous T-cell lymphoma, CTCL). Repeated peripheral blood examinations showed an increased percentage of lymphocytes of the helper phenotype. A bone marrow examination revealed a ratio of suppressor/helper subsets of 1:4. The findings in the skin seem to be inconsistent with most of the results of previous studies in patients with CTCL; the significance of these findings is discussed.This study was partly supported by the Deutsche Forschungsgemeinschaft, Grant no. Lo 285/2-1This work is dedicated to Prof. Th. Nasemann on occasion of his 60th birthday     

The immunopathology of hypopigmented mycosis fungoides

Two dark-skinned patients presented with well-defined hypopigmented macules, a biopsy of which showed the characteristic features of mycosis fungoides. Immunohistochemical studies with UCHT1 antiserum (a pan T cell marker) confirmed the almost complete T lymphocyte nature of the infiltrate. UCHT4 antiserum (a suppressor/cytotoxic marker) showed that the epidermal infiltrate consisted predominantly of T suppressor/cytotoxic cells and Leu3a antiserum (a T helper cell marker) showed that the dermal infiltrate was composed of helper and suppressor/cytotoxic cells in approximately equal proportions. Ultrastructural studies identified normal melanocytes in the basal layer of the epidermis from both patients but mitochondrial vacuolation was seen occasionally in Case 2. We believe that this is the first report which documents T cell subsets in skin biopsies from patients with hypopigmented mycosis fungoides. The finding of a predominance of suppressor/cytotoxic T lymphocytes in the epidermal infiltrate of these two patients contrasts with the usual predominance of T helper lymphocytes in the more common forms of mycosis fungoides.     

Clinical and The use of a double-label immunoperoxidase monoclonal antibody technique in the investigation of patients with mycosis fungoides

Using a double-labelling immunoperoxidase and monoclonal antibodies raised against T helper and T suppressor cells and Langerhans cells, we have found that the clinically involved skin of seventeen mycosis fungoides (MF) patients shows an increase in number of Langerhans cells which are in contact with T helper lymphocytes. T suppressor lymphocytes are also present, but are generally seen singly situated at a distance from the T helper/Langerhans cell clusters.     

Immunophenotyping of the eczematous flare-up reaction in a nickel-sensitive subject

An eczematous flare-up reaction, occurring at a previously involved site, which followed oral challenge with 5.6 mg of nickel in a 29-year-old nickel-sensitive woman, was biopsied and studied by immunohistochemistry. The cellular infiltrate in the dermis and epidermis at 8 days was predominantly of Leu 3a phenotype (helper/inducer T lymphocytes), with smaller numbers of Leu-2a-reactive (suppressor/cytotoxic) T lymphocytes. Many infiltrating cells were DR-positive. No increase in epidermal Leu-6-positive Langerhans cells was seen but Leu-6-reactive cells were noted in the dermal infiltrate. Keratinocytes showed some expression of class II antigen (mainly DR). In comparison with the 48-hour allergic patch test reaction, the eczematous flare-up site showed no increase in epidermal Langerhans cell numbers nor infiltration with macrophages, but the responses were similar since both showed a superficial T cell reaction in the skin.     

In situ identification of mononuclear cells in lichen planus

In this study, the in situ immunological typing of cell populations in lichen planus was attempted. T lymphocytes and suppressor/cytotoxic subsets, B lymphocytes, macrophages, immunocytes and Langerhans' cells were studied by one or more technical parameters and semiquantitative assessment of T cell populations were carried out. A critical evaluation of assays for T cell characterization was also attempted. T cells were found predominant in lichen planus infiltrate but macrophages were also many. Langerhans' cells were increased in the epidermis compared to normal skin and contact dermatitis.     

An analysis of infiltrating cells in human ringworm

Immunohistochemically we examined six patients with ringworm which exhibited a well-marked erythematous edge and central resolution. To obtain a panoramic view of the immunopathological features of various evolutionary stages of skin lesion, we took a radial slice biopsy specimen extending from normal-appearing skin to central resolution. The actively involved edge had a moderate amount of dermal infiltrate of mononuclear cells which mostly consisted of T cells. The ratio of the helper T cells to suppressor T cells was 3.4. At the central resolution, the dermal infiltrate was mild, and the helper T/suppressor T ratio was 3.0. The well-marked erythematous edge showed a decrease of OKT6 positive Langerhans' cells in the epidermis, while many OKT6 positive cells were present in the upper dermis. At the central resolution, however, OKT6 positive cells increased in number in the acanthotic epidermis.     

Langerhans cells in human warts

Seventy-six warts (15 plantar, 38 hand, 16 miscellaneous and seven anogenital lesions) taken from 55 patients, were studied by indirect immunofluorescence with monoclonal antibodies specific for T-cell subsets, Langerhans cells (LC) and HLA-DR antigen. The results were related to the presence of viral antigen. Approximately 80% of the lesions showed an infiltrate. Only 19 lesions contained helper/inducer or suppressor/cytotoxic T cells. The distribution of LC was abnormal in 65% of biopsies which contained LC in the dermis, and 29% were devoid of LC in the epidermis. Many lesions had reduced numbers of LC in the epidermis. The disappearance of LC from the epidermis was related to the presence of viral antigen, but not to the presence of particular T-cell subsets. Infiltrating cells were sometimes HLA-DR-positive, whereas basal cells did not express HLA-DR antigen, irrespective of the density of the infiltrate.     

Effectiveness of cyclosporine treatment in severe psoriasis: a clinical and immunologic study

The effectiveness of low doses of cyclosporine (3 to 5 mg/kg/day) for short-term treatment in 13 patients with severe psoriasis was studied. The psoriasis cleared in 12 of 13 patients within 3 to 4 weeks of treatment, and there was appreciable improvement in the thirteenth patient. No major side effects were observed: two patients showed biochemical evidence of slight transient renal dysfunction and three others had cutaneous infections (two viral and one mycotic). An immunohistologic study showed that the psoriatic plaques contained an infiltrate composed mainly of activated helper T lymphocytes. After 15 days of cyclosporine treatment, CD4+ cells were significantly fewer in the epidermis and dermis, and Langerhans cells were more regularly distributed in the epidermis. Our studies of neutrophil chemotaxis showed that it is not significantly influenced by cyclosporine in vitro but is decreased in vivo.     

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.     

Tissue and blood T-lymphocyte subpopulations in erythema nodosum leprosum

To study T lymphocytes in erythema nodosum leprosum (ENL), monoclonal antibodies were used to identify T-lymphocyte subpopulations in the blood and skin lesions of patients with ENL and patients with nonreactional lepromatous leprosy. The blood of nonreactional lepromatous patients had a lymphopenia and a proportionate reduction in pan T cells, helper-inducer, and suppressor-cytotoxic subsets, but a normal helper-suppressor ratio, as compared with controls. Patients with ENL did not differ significantly from the controls. In skin lesions, an admixture of helper and suppressor phenotypes among foamy histiocytes was found. The ENL tissue had more numerous cells of the helper-inducer phenotype and fewer of the suppressor-cytotoxic phenotype, as compared with nonreaction lepromatous tissues. In 22 patients with simultaneous examination of tissue and blood T-cell subsets, there was no correlation between tissue and blood helper-suppressor ratios, indicating that some sort of selection process brings lymphocytes into tissues from peripheral blood.     

Immunophenotyping of skin cells during healing of suction blister injury.

The time course of appearance and the dynamic changes of immunocompetent cells were assessed in human skin following sterile suction blister would healing. During epidermal regeneration, a local increase in Langerhans cells (LC) and the appearance of a mononuclear cell infiltrate were observed. Initially, T cells were exclusively of the T helper/inducer subtype, whereas during the later stages of the healing process an increasing number of cytotoxic/suppressor T cells (up to 30%) was observed. Keratinocytes of neither the adjacent nor the newly formed epidermis expressed HLA-DR over the course of the wound-healing process. Our results suggest that immune-competent cells such as T cells and LC may play an important role in the regulation of the wound-healing process.     

Fc epsilon R11/CD23 receptor distribution in patch test reactions to aeroallergens in atopic dermatitis.

There is increasing evidence that exposure to organic allergens may induce or exacerbate lesional skin in patients with atopic dermatitis. In this study, patients with atopic dermatitis were patch tested to 11 common organic allergens and to control chambers containing 0.4% phenol and 50% glycerin in 0.9% saline. In biopsies from positive patch test reactions, patch test control skin, lesional eczematous and non-lesional skin from atopic individuals, and normal skin from non-atopic volunteers, the presence and distribution of macrophages (RFD7+), dendritic cells (RFD1+), and Langerhans cells, and the expression of the low-affinity receptor for IgE (CD23) were investigated. In patch test reactions and lesional skin samples, inflammatory infiltrates of diffusely distributed macrophages (RFD7+), dendritic cells (RFD1+), T lymphocytes (RFTmix+), and Langerhans cells (CD1+) were seen, the latter being present in both the epidermis and the dermis. The numbers of Langerhans cells were reduced in the epidermis and increased in the dermis in patch test reactions and lesional skin compared to their controls. Double staining revealed a change in the distribution of CD23 antigen. In patch test control and non-lesional biopsies many macrophages and only a few Langerhans cells within the dermal infiltrates expressed this antigen. In patch test reaction and lesional skin samples, however, the proportion of CD23+ dermal Langerhans cells had increased compared to macrophages. Furthermore, in these latter samples an increased proportion of dermal CD1+ cells expressed the dendritic cell (RFD1+) marker. These results show that following antigen challenge there are marked similarities between the phenotype of the cellular infiltrate in patch test reaction and lesional skin biopsies, and also demonstrate a changing distribution of CD23 on antigen-presenting cells.     

Changes in Lymphocyte and Langerhans Cell Populations in Allergic and Irritant Contact Dermatitis

We observed in situ changes in lymphocyte subpopulations and Langerhans cells during allergic and irritant contact dermatitis using immunohistochemical staining methods with monoclonal antibodies to cell surface antigens. In both types of contact dermatitis, there was a perivascular infiltrate of T lymphocytes, with helper/inducer T cells predominating. B cells were absent, and natural killer cells were absent or sparse. During the course of allergic contact dermatitis, Langerhans cells showed a striking sequential change in location, with the cells first in the epidermis, then perivascularly in the dermis (days 1-14), and returning to the epidermis (days 14-21). In irritant contact dermatitis, the Langerhans cells were initially identified in the epidermis and then appeared diffusely in the dermis (days 1-2). The numbers in the dermis then decreased abruptly (day 4). They were again identified in normal numbers in the epidermis (day 21). The response of Langerhans cells appears to be different between allergic and irritant contact dermatitis.     

Lymphocyte subsets and Langerhans cells/indeterminate cells in erythema multiforme

A peroxidase-antiperoxidase study using monoclonal antibodies directed against T and B lymphocytes and Langerhans cells/indeterminate cells (LC/IC) was undertaken in order to understand more clearly the changes observed in erythema multiforme. At the various stages of development, from normal skin to target lesions, the quantity of inflammatory cells differed, but in each case the number of T8+ (cytotoxic/suppressor) cells was greater than the number of T4+ (helper/inducer) cells in the epidermis, whereas the latter exceeded the former in the dermis. Concomitant with the initial epidermis changes, there was an increase in the number of T6+ (LC/IC) cells in the upper and lower epidermis. With slight to moderate basal unit destruction, the number of LC/IC in the upper epidermis exceeded those in the lower epidermis. With severe basal unit destruction, there was a loss of LC/IC in the lower epidermis as detected by T6 reactivity. In fully formed blisters, the LC/IC in the upper half of the epidermis were decreased in parallel with the degree of epidermal necrosis. The character of the lymphocytic inflammatory infiltrate and redistribution in LC/IC are similar to those findings described in allergic contact dermatitis. The clinical, histologic, and immunopathologic changes in erythema multiforme appear to be due in part to cellular immune mechanisms with the lymphocyte as the predominant effector cell, and our data suggest a possible role for LC/IC in this disorder.     

Erythema chronicum migrans: evidence for cellular immune reaction in the skin lesion

Skin biopsy specimens from 9 patients with erythema chronicum migrans (ECM) were studied immunohistochemically using a series of monoclonal antibodies. In biopsy specimens taken from the erythematous peripheral portion of ECM the perivascular infiltrates were composed predominantly of LEU-4+ T cells. LEU-3a + helper/inducer T cells were more numerous than LEU-2a + cytotoxic/suppressor T cells. Of particular interest was the high number of LEU-6+ Langerhans cells in the epidermis and dermis of specimens taken from the erythematous portion of ECM as well as from the noninflammatory skin outside the erythema. The presence of LEU-6+ Langerhans cells and T cells in the ECM lesions suggests that, apart from humoral factors, a cell-mediated immune response directed against Borrelia burgdorferi antigen is important as well in the pathogenesis of this disease.     

Morphological and immunohistochemical study of immediate contact dermatitis of the hands due to foods

This histological and immunohistochemical study of 6 food handlers affected by immediate contact dermatitis due to foods shows that apparently normal skin of patients with this condition presents several histological and immunohistochemical abnormalities. Skin biopsies of normal hand skin showed focal parakeratosis and moderately dense dermal infiltrates. Immunohistochemistry showed an increased number of Langerhans cells in the epidermis and in the superficial dermis and a mononuclear dermal infiltrate consisting of peripheral T lymphocytes with a CD4/CD8 ratio of 5-6/1. Biopsies of the immediate vesicular reactions induced by foods showed spongiotic vesicles within the epidermis and a moderate to dense mononuclear dermal perivascular infiltrate. The immunohistochemical features were similar to those described in apparently normal skin. The mechanism of this immediate vesicular reaction requires further research. The rapid appearance of the lesions (after 20-30 min) probably excludes an immunological cell-mediated pathogenesis. A non-immunological mechanism due to direct liberation of mediators by foods is more readily conceivable than an immediate immunological type of contact reaction.     

Distribution of natural killer cells and lymphocyte subclasses in Jessner's lymphocytic infiltration of the skin and in cutaneous lesions of discoid and systemic lupus erythematosus

We studied the cell infiltrates in biopsies from lymphocytic infiltration of the skin (LIS), with six monoclonal T cell antigen-specific antibodies and compared the reactivity pattern with those in biopsies from discoid and systemic lupus erythematosus skin lesions and allergic contact skin reactions. A newly described antibody (NK₉) recognizing natural killer (NK) cells and activated cytotoxic T lymphocytes was included, and the numbers and activity of circulating NK cells was determined. Immunohistochemical staining revealed that the numbers of NK₉-positive cells were highest in LIS. The distribution of T lymphocytes (OKTii + ve), helper T cells (OKT₄+ ve), suppressor T celts (OKT8 + ve), Langerhans cells (OKT6 + ve) and activated T cells (anti-Tac + ve) in LIS differed from those in DLE, SLE and allergic contact reactions. However, the number of circulating NK cells (large granular lymphocytes) and the NK activity in peripheral blood were normal in LIS. We conclude that in LIS a distinct type of T cell activation occurs; the cause of this remains to be determined.     

Quantitation of cutaneous Langerhans cells of sarcoidosis patients

Langerhans cells play a role in cell-mediated immune reactions which are often depressed in sarcoidosis. We examined the epidermis of 17 anergic patients with sarcoidosis (Kveim-reactive and/or biopsy-proved) for the number of Langerhans cells in noninvolved skin and in any cutaneous sarcoidal lesions. Skin biopsies of 10 healthy volunteers served as controls. In comparison to controls, the epidermis overlying noninvolved (p less than 0.05), sarcoidal (p less than 0.0005), and Kveim-reactive (p less than 0.005) skin contained significantly fewer detectable Ia and T6 antigen-bearing Langerhans cells. The reductions within noninvolved skin were most pronounced in patients with multisystem disease. Lower epidermal Langerhans cell densities, in comparison to controls, were detected in both prednisone-treated and untreated patients. Epidermis overlying sarcoidal skin of untreated patients contained significantly fewer Ia and T6 antigen-bearing Langerhans cells (p less than 0.05, p less than 0.0025, respectively) than epidermis from noninvolved skin. Whether reduced numbers of cutaneous Langerhans cells are due to either a local and/or systemic effect of sarcoidosis, or reflect the anergic state of these patients is unknown.