Lichen planus and chronic graft-versus-host reaction. In situ identification of immunocompetent cell phenotypes

The purpose of the present study was lo examine the phenotype of the cutaneous immunocompetent cells in lichen planus and chronic graft versus host (GVH) reaction infiltrates, by the use of monoclonal antibodies directed against T cell populations and Langerhans cells.
Our results in lichen planus suggest an immunological reaction similar to the delayed hypersensitivity reaction, including all the immunocompetent cell sub-populations, with a first stage of antigenic information by Langerhans cells (OKT6 +, BL6 +, HLA-DR+) and helper cells, and a second stage mediated by suppressor/cytotoxic cells.
The results from the study of GVH reaction also suggest an effect mediated by suppressor/cytotoxic cells (OKT3+, OKT4−, OK.T8+, HLA-DR+).
Our results favour the existence of a lymphocytotoxic process in lichen planus and chronic GVH reaction.     

Different in situ distribution patterns of dendritic cells having Langerhans (T6+) and interdigitating (RFD1+) cell immunophenotype in psoriasis, atopic dermatitis, and other inflammatory dermatoses

Dendritic cells bearing Langerhans cell (OKT6+) or interdigitating cell (RFD1+) immunophenotype may be regularly detected within the dermis of chronic skin diseases characterized by a lymphohistiocytic (lymphoreticular) infiltrate. These 2 subsets of antigen-presenting cells within the dermis of lesions of exacerbating chronic plaque psoriasis, exacerbating nummular dermatitis (discoid eczema), atopic dermatitis, allergic contact dermatitis, pityriasis rosea, lichen ruber planus, and cutaneous lupus erythematosus were quantified using computer-assisted morphometry. The mean dendrite length per dermal dendritic cell was significantly higher for RFD1 than for OKT6 (74.4 +/- 0.98 microns vs 70.0 +/- 1.26 microns: p = 0.0023). The mean dendrite length per dermal dendritic cell was remarkably constant for each marker in the various diagnostic categories studied. Disease-specific patterns of total dendrite length and number (expressed per 100 infiltrating mononuclear cells) of these 2 dendritic cell types within the subepidermal infiltrates were obtained. Pityriasis rosea was characterized by its unique high percentage of OKT6+ Langerhans cells. Atopic dermatitis and psoriasis had relatively high percentages of both RFD1+ interdigitating cells and OKT6+ Langerhans cells. Nummular dermatitis had an intermediate number and total dendrite length for OKT6, but was relatively low in RFD1+ cells. Allergic contact dermatitis, lichen planus, and lupus erythematosus had low numbers and dendrite lengths for both dendritic cell subsets. It is suggested that pityriasis rosea is characterized by an abnormal migration pattern of Langerhans cells. Psoriasis and atopic dermatitis may be examples of diseases in which skin-localized antigen-presenting and T-cell-inducing events are continuously taking place. The other diseases may reflect inflammatory processes in which local antigen presentation is less relevant to the tissue reaction.     

Anti-epidermal cytotoxic reactions in the effector stage of lichen ruber planus

Immunohistochemical investigations on skin lesions of lichen ruber planus indicate HLA-DR-expression by keratinocytes, Langerhans cells (LC), and infiltrated mononuclear cells. In the early stage of this condition, the lymphocytic infiltrate consists of a considerable part of (CD8+) cytotoxic/suppressor- and (LGL+) natural killer cells. Evidently, the cytotoxic potential of these lymphocyte subpopulations is directed against basal keratinocytes and epidermal LC. As a result of cytotoxic/cytolytic activities, non-cell associated CD 1-reactivity accumulates in the dermoepidermal junction zone, thus indicating damaged LC.     

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.     

Subpopulations of mononuclear cells in microscopic lesions of psoriatic patients. Selective accumulation of suppressor/cytotoxic T cells in epidermis during the evolution of the lesion

The age of microscopic lesions in psoriatic subjects was assessed from the stacking characteristics in the horny layer and related to type and density (cells/tissue volume) of mononuclear cells in the epidermis and the dermis determined by immunoperoxidase methods using monoclonal antibodies. Pan T cells (Lyt-2+, Lyt-3+, Leu-4+, OKT3+), T helper cells (Leu-3a+, OKT4+), T suppressor/cytotoxic cells (Leu-2a+, OKT8+), Ia+ cells and monocytes (OKM2+, BRL alpha mono+) were determined in epidermis and dermis. The psoriatic lesion was divided into regions underneath a parakeratotic and an orthohyperkeratotic/hypergranular portion of the horny layer and contrasted with perilesional and uninvolved psoriatic skin as well as with healthy skin. In the various regions and skin layers, the cell density was highest in parakeratosis and decreased toward normality with decreasing histologic abnormality. The relation between epidermal and dermal cell densities of the T-cell subsets was modified in the involved psoriatic skin with a selective preponderance of T suppressor/cytotoxic cells in the epidermis. The accumulation was present in the youngest lesion found (3 days) and cell densities were unchanged in older lesions. The findings suggests that the altered relationship in the subsets of T cells has an important role during the induction and progress of the psoriatic process in the skin.     

The role of perforin-mediated apoptosis in lichen planus lesions

Lichen planus is recognized as a T-cell-mediated disease. Histologically, it is characterized by the formation of colloid bodies representing apoptotic keratinocytes. The apoptotic process mediated by CD8⁺ cytotoxic T lymphocytes (CTLs) and NK cells mainly involves two distinct pathways: the perforin/granzyme pathway and the Fas/FasL pathway. So far, little is known regarding the role of perforin-mediated apoptosis in lichen planus. In the present study, the expression and distribution of perforin, T and NK cell subsets in the epidermis and dermis of lesional and nonlesional lichen planus skin were studied. Skin biopsy specimens from lesional and nonlesional skin of ten patients with lichen planus and eight healthy persons were analysed by immunohistochemistry. Significant accumulation of T cells, particularly of CD4⁺ and CD8⁺ subsets, was found in both epidermis and dermis of lichen planus lesions compared with nonlesional and healthy skin. There were no significant differences in the incidence of NK cells (CD16⁺ and CD56⁺) between lesional, nonlesional and healthy skin. Perforin expression was significantly upregulated in the epidermis of lichen planus lesions. In conclusion, accumulation of perforin⁺ cells in the epidermis of lichen planus lesions suggest a potential role of perforin in the apoptosis of basal keratinocytes.     

A quantitative assessment of Langerhans cells in oral mucosal lichen planus and leukoplakia

Quantitative analysis of Langerhans cells stained with OKT6 in the epithelium of 42 patients with oral mucosal lichen planus and 14 patients with oral mucosal leukoplakia showed significantly greater numbers of Langerhans cells in lichen planus. This may reflect the role of Langerhans cells in the pathogenesis of this condition and their identification could help in distinguishing lichen planus from leukoplakia.     

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.     

Immunohistologic identification of antigen-presenting cells in cutaneous sarcoidosis

Considerable evidence exists to show that activated T lymphocytes preferentially accumulate at sites of disease activity in sarcoidosis. Langerhans cells, which can be recognized by reactivity with an antibody to the T6 antigen are thought to play a primary role in T-lymphocyte activation by the skin, a tissue frequently involved in sarcoidosis. This immunohistologic study examined the distribution of OKT6-positive cells and surface expression of HLA-DR antigen in cutaneous sarcoid lesions. Skin specimens stained with an anti-HLA-DR antibody demonstrated diffuse staining of the granulomas. In addition, keratinocytes, which do not normally express HLA-DR antigens, were found to stain with monoclonal antibody to HLA-DR in an intercellular pattern. Examination of specimens for OKT6-reactive Langerhans cells revealed significantly greater concentrations in the epidermis overlying sarcoidal granulomas (33 +/- 7 cells/mm) than in the epidermis of age-, sex-, and race-matched controls (11 +/- 3 cells/mm, p less than 0.001). Of greater importance was the demonstration that significant numbers of OKT6-positive cells were present within the dermal sarcoid granulomas (19-208/mm2) in a distribution that paralleled that of Leu-3a-positive T lymphocytes. These data suggest that the epidermis may participate in activation of lymphocytes in cutaneous sarcoidosis, and implicate OKT6-positive cells in granuloma formation.     

T cell subsets and Langerhans cells in lichen planus: in situ characterization using monoclonal antibodies

Skin biopsies from four patients with lichen planus were studied using monoclonal antibodies directed against T lymphocytes. Anti-T1 and anti-T3 antibodies, which react with all peripheral T cells, stained most cells in the dermal infiltrates. The majority of infiltrating cells also stained with anti-T4 and anti-T4b antibodies, which react with helper/inducer cells, whereas a minority of cells stained with anti-T8 antibody, which reacts with cytotoxic/suppres-sor cells. Surface IgM was not identified on any infiltrating cells, providing evidence against B cell participation. Intraepidermal and dermal cells with long cytoplasmic extensions stained with anti-T6 antibody in all cases, defining them as Langerhans cells or their precursors. T6-positive cells were seen in greater number than in normal control epidermis and dermis. The results indicate that well-developed lesions of lichen planus are characterized by an influx of helper/inducer T lymphocytes and increased numbers of Langerhans cells. These observations support the contention that cellular immunity is important in the pathogenesis of this disorder.     

Quantitative studies on nonlymphoid mononuclear cell subpopulations in cutaneous infiltrates. I. Stage-related changes of dendritic nonlymphoid mononuclear cells, Langerhans' cells, and macrophages in lichen planus lesions

In previous investigations on lichen planus, we suggested that in early lesions T4-positive cells might be antigen-specifically driven, whereas in late lesions T8-positive cells may be cytotoxic to keratinocytes. To verify this hypothesis, we investigated the following nonlymphoid mononuclear cell subpopulations in early versus late lichen planus lesions: interdigitating cells (phenotype: S100-positive, lysozyme-negative, T6-negative, M3-negative), Langerhans cells (phenotype: S100-positive, lysozyme-negative, T6-positive, M3-negative), macrophages (phenotype: S100-negative, lysozyme-positive, T6-negative, M3-positive). Interdigitating cells were moreover identified in semithin and ultrathin sections by distinctive morphological characteristics. The S100-positive/lysozyme-positive cell ratio was higher (p less than 0.01) in early lesions than late lesions. In dermis but not in epidermis (NS), of early lesions, T6-positive cells were less represented than S100 positive cells (p less than 0.025). Thus, Langerhans' cells largely predominated over interdigitating cells in epidermis, but the two populations were both represented in dermis. Lysozyme-positive and M3-positive cells, more abundant in late lesions than in early lesions (p less than 0.001), were often filled with pigment granules.     

Macrophage — T-lymphocyte interaction in lichen planus

Summary Papular lichen planus lesions from 12 patients were studied by a double-step immunocyto-chemical method to detect T-lymphocytes. Semithin sections were studied by light microscopy and ultrathin sections examined by electron microscopy. In the dermal infiltrate, many T-lymphocytes appeared closely juxtaposed to macrophages or Langerhans cells, frequently arranged in a rosette-like pattern. In the epidermis, T-lymphocytes were juxtaposed to macrophages or Langerhans cells and to degenerated keratinocytes. The close relationship between T-lymphocytes, macrophages or Langerhans cells and degenerated keratinocytes supports the hypothesis that lichen planus is immunological in nature: T-lymphocytes, after interacting with macrophages or Langerhans cells, become cytotoxic for keratinocytes.Supported in part by the Consiglio Nazionale delle Ricerche; Roma, Italia, grant no 790181604The ultrastructural findings have been presented by prof. Giannotti in occasion of the VII European Meeting of the SCUR Vienna, 8–10 May 1980     

Immunopathological aspects of etretinate therapy in lichen planus

Biopsy specimens of involved and uninvolved skin were studied in 8 patients suffering from generalized lichen planus (LP) before and after successful etretinate treatment using murine monoclonal antibodies against several cell surface markers of effector and/or accessory cells of the immune system. In contrast to untreated LP-lesions, in biopsies obtained after etretinate therapy from healed involved skin a markedly disintegrated HLA-DR expression on keratinocytes and intraepidermal Langerhans cells and a nearly total loss of suppressor/cytotoxic T lymphocytes at the dermo-epidermal interface were found. Our data emphasize the key position of the class II antigen expression on various immunocompetent cells, as a marker for a cell-mediated immune reaction, on the other hand, stress the involvement of keratinocytes, Langerhans cells and suppressor/cytotoxic T lymphocytes in the pathogenesis of LP.     

Lymphocytes and Langerhans cells in patch tests

The distribution of immunocompetent cells was analysed in allergic (nickel) and irritant (dithranol) patch tests using conventional transmission electron microscopy and labelling with monoclonal antibodies in an avidin-biotin immunoperoxidase study. The biopsies were taken 24 or 48 h after the allergen/irritant application. In allergic and irritant reactions, most inflammatory cells were OKT11 positive (pan T lymphocytes). The majority of these cells were also OKT4 positive (helper/inducer T lymphocytes), while the minority were OKT8 positive (suppressor/cytotoxic T lymphocytes). NK9 positive cells (natural killer cells) were observed in small numbers. The number of dendritic OKT6 and OKIal positive cells (Langerhans cells) in the epidermis was unaffected in allergic reactions. In irritant reactions, a normal number of OKT6 positive Langerhans cells was observed, while the number of OKIal positive cells had increased in the epidermis. Dithranol caused prominent fine structural changes in the mitochondria of the Langerhans cells, while the keratinocytes appeared largely unaffected. The present study indicates that allergic and irritant patch tests cannot be differentiated reliably using current immunohistopathological or electron microscopic techniques, in spite of the small differences observed.     

Expression of OKM5 antigen on epidermal cells in mycosis fungoides plaque stage

To characterize and quantitate potential antigen-presenting cell subsets in the epidermis of patients with cutaneous T-cell lymphoma, epidermal cells in suspension were obtained from involved and uninvolved skin. Involved epidermis contained increased numbers of OKT6+HLA-DR+ Langerhans cells and a variable number of OKM5+ epidermal cells (ECs) in all mycosis fungoides (MF) patients tested (N = 14). The OKM5+ EC population from involved epidermis of MF patients were heterogeneous and comprised both OKM5+HLe1- keratinocytes and OKM5+HLe1+ leukocytes. Uninvolved epidermis, in 6 of 14 patients with MF, contained a small number of OKM5+ leukocytes; however, no OKM5+ keratinocytes were detected. Neither OKM5+ leukocytes nor OKM5+ keratinocytes were detected in the epidermis obtained from healthy controls. The increased number of potential antigen-presenting cells, that is, OKT6+HLA-DR+ Langerhans cells and OKM5+HLA-DR+ monocytic leukocytes, in the epidermis of patients with MF may be important for the activation of abnormal T cells contained within the epidermis of these patients. Such activated T cells may release gamma-interferon and induce expression of both HLA-DR and OKM5 antigens on keratinocytes. OKM5+ keratinocytes are present in the epidermis of patients with MF, but not in normal skin, and may thus play a role in the pathogenetic mechanisms of mycosis fungoides by recruitment of immunocompetent cells to the epidermis.     

Heterogeneity of dermal OKT6+ cells in inflammatory and neoplastic skin diseases

This immunopathologic study of both normal and pathologic skin specimens (contact dermatitis [CD], lichen planus [LP], cutaneous T cells lymphoma [CTCL], and histiocytosis X [HX]) allowed as to differentiate four types of dermal OKT6+ cells: (1) cells with the same morphologic features as epidermal Langerhans cells (LCs), rarely found in either normal or pathologic dermis; (2) cells structurally similar to LCs but lacking Birbeck granules (BGs), found mainly in CD and LP; (3) larger cells rich in cytoplasmic organelles, only 5% of which contained BGs. They were especially common CTCL; and (4) cells typical of HX.     

Characterization of cutaneous infiltrates in MRL/lpr mice monitored from onset to the full development of lupus erythematosus-like skin lesions.

The skin is a primary site injured in lupus erythematosus (LE), but it is still controversial whether the injury is due to cells of the mononuclear infiltrate and which immunocompetent cells play the major role in the development of cutaneous LE. To better characterize the role of immunocompetent cells, we performed an immunohistochemical examination of these cells in LE-like skin lesions in MRL/Mp-lpr/lpr (MRL/lpr) mice. Skin lesions in 60 female MRL/lpr mice were monitored from onset to full development. Skin specimens from each stage were stained for epidermal Ia+ Langerhans cells (Ia(+)-LC), for Thy-1+ dendritic epidermal cells (Thy-1+DEC), and for the phenotype of the mononuclear cell infiltrates. The numbers of Ia(+)-LC and Thy-1+DEC were decreased markedly in the skin lesions at the later stage. However, the numbers of Ia(+)-LC were increased significantly in the central portion of lesions at an early stage and in the peripheral portion of lesions later. L3T4+ cells were predominant, and the L3T4/Lyt-2 ratio was high in dermal infiltrates at an early stage. With advancing stage, the L3T4/Lyt-2 ratio gradually decreased in dermal infiltrates, whereas the Thy-1.2/Lyt-2 ratio in lymph nodes was reversed. L3T4+ cells were especially predominant in dermal infiltrates under the epidermis with increased numbers of Ia(+)-LC. This immunohistochemical analysis of a mouse model of cutaneous LE revealed changes in immunocompetent cell populations with the evolution of skin lesions, and we conclude that Ia(+)-LC and Thy-1+DEC, as well as L3T4+ and Lyt-2+ cells, may play pathogenic roles in the development of skin lesions.     

Oral lichen planus: an immunoperoxidase study using monoclonal antibodies to lymphocyte subsets

Sixteen biopsies from patients with oral lichen planus (10), simple keratosis (3) and lichenoid reactions (3) were studied using monoclonal antibodies directed against lymphocyte markers. T lymphocytes predominated in the cellular infiltrates of the epithelium and lamina propria of all biopsies, and cells positive with Leu 3a and Leu 2a antibodies were also present at both sites in all specimens. However, many of the Leu 3a positive intraepithelial cells had a dendritic appearance and distribution consistent with their being Langerhans cells. Limited results from cell counts on three lichen planus specimens suggested that the majority of intraepithelial T cells were of the cytotoxic/suppressor phenotype (OKT3 and Leu 2a positive).     

Demonstration of a subpopulation of Ia+ T-helper cells in mycosis fungoides and the Sézary syndrome

This is a report of the finding of a T-cell subpopulation bearing T-helper cells and Ia antigens in specimens of skin from patients with mycosis fungoides and the Sézary syndrome. Frozen sections of skin tissue from eight patients examined with monoclonal antibodies against mature T-cells, helper T-cells, suppressor T-cells, and Ia antigens exhibited similar staining patterns by a modified immunoperoxidase method. Antibodies against mature T-cells and helper T-cells stained 70-80% of the lymphocytes in the dermis. The antibody defining the phenotype of suppressor T-cells labelled 5-10% of the lymphocytes scattered throughout the lesions. Eighty to 90% of the lymphocytes took the stain for Ia antigen. Anti-thymocyte antibody (OKT6) stained cells in both the epidermis and dermis of the specimens. Of nonmalignant conditions examined, lesions from five cases of lichen planus exhibited a quantitatively different staining pattern than that of mycosis fungoides in that the number of T-helper cells was about equal to the number of T-suppressor cells. The findings reported are evidence for a homogeneous population of T-helper, Ia-positive lymphocytes in the cutaneous lesions of mycosis fungoides and the Sézary syndrome.     

T cell subsets and macrophages in lichen planus. In situ identification using monoclonal antibodies and histochemical techniques

Skin lesions from 6 patients with lichen planus were studied for the presence of T cells and T cell subsets using monoclonal antibodies and indirect immunoperoxidase technique. Small numbers of Leu-2a-reactive suppressor-cytotoxic cells were present early in the basal cell layer in 2 patients with recent lesions. The analysis of T cell subsets revealed the predominance of anti-pan-T-cell (Leu-1)- and Leu-3a-reactive helper-inducer cells in 4 patients with older active lichen planus lesions. Significant numbers of suppressor-cytotoxic cells were observed in the papillary dermis and within the epidermis associated with hydropic degeneration of the basal cell layer. Activated T lymphocytes with focal acid phosphatase activity, together with activated histiocytes-macrophages with strong diffuse activity of acid phosphatase and non-specific esterase, were identified in the dermis and within the epidermis. These findings suggest that a cytotoxic immune process directed against the basal cell layer of the epidermis is the dominant pathogenic event in lichen planus.