单克隆抗体对异位性皮炎损害内浸润细胞免疫荧光表现型的鉴定

应用冰冻切片下间接免疫荧光技术及针对总T细胞、T辅助/诱导细胞及T抑制/杀伤细胞的单克隆抗体,对一例异位性皮炎皮损的浸润细胞进行组织内免疫荧光表现型鉴定.结果表明,该例异位性皮炎损害内的浸润细胞,大部份有T辅助细胞的表面标志.     

In situ characterization of tissue lymphoid cells of cutaneous infiltrates using specific membrane antigens

A method for identification of B and T lymphocytes on frozen tissue sections is described. The technique was initially established on mouse spleen. Thymodependant cells were identified by anti-Thy-i and anti-theta (alloantigens specific for mouse T lymphocytes) sera and thymoindependant cells by Fab-Po anti-Ig serum. This technique was subsequently applied to a study of benign and malignant cutaneous lymphoid infiltrates. Specific membrane markers, human B-lymphocyte antigens (HBLA) for B lymphocytes and human T-lymphocyte antigens (HTLA) for T lymphocytes, were revealed by peroxidase immunolabelling. A prominent T-Iymphocyte population was observed in the cutaneous lesions of sarcoidosis, lichen planus, halo naevus and lupus erythematosus. In contact dermatitis, the prominence of the T-cell infiltrate is in accordance with the delayed hypersensitivity reaction. Immunoenzymatic labelling of sections of epidermotropic malignant cutaneous lymphoma confirmed the T-cell nature of the intraepidermal neoplastic cells. This technique of direct specific labelling on sections should also be applicable to electron microscopy.     

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.     

In situ characterization of cellular infiltrates in lupus vulgaris indicates lesional T-cell activation.

Skin biopsy specimens from nine patients with lupus vulgaris were examined in situ by means of monoclonal antibodies directed against phenotypes of lymphocyte subsets, Langerhans cells, HLA-DR antigens, and interleukin 2 receptor. The epidermis showed prominent changes, including intense expression of HLA-DR on keratinocytes, increase in epidermal cell layers, moderate to high Langerhans cell hyperplasia, and infiltration by CD3+ pan-T cells as well as CD8+ (cytotoxic/suppressor) and CD4+ (helper/inducer) T cells. The predominant lymphocyte in the dermal granulomas was the activated CD3+ T cell, expressing major histocompatibility complex class II antigens and interleukin 2 receptor. CD4+ and CD8+ cells were randomly distributed among the epithelioid cells, which showed intense staining for major histocompatibility complex class II antigens. In all except two patients, the CD4+ population was greater than that of the CD8+ cells. CD1+ Langerhans cells were scattered in moderate numbers in the dermal granulomas. Acid-fast bacilli were conspicuously absent in the biopsy specimens. These features suggest that T-cell activation and Langerhans cell hyperplasia are prominent features of dermal tuberculosis.     

Characterization of benign cutaneous lymphocytic infiltrates by monoclonal antibodies

Skin biopsies from nine patients with a histological and/or clinical diagnosis of cutaneous lymphocytoma, lymphoplasia or Jessner's lymphocytic infiltrate were examined by immunoenzymatic labelling with a panel of monoclonal antibodies against lymphocytes and accessory cells. Similar cellular constituents were demonstrated in the biopsies from three patients with lymphocytoma, two with lymphoplasia and two with atypical lymphocytes, but a protracted benign clinical course. The infiltrates from these patients consisted of T cells, Langerhans cells, related HLA-DR positive dendritic dermal cells and clusters of polyclonal B cells. In four patients, the B cell clusters contained B cell accessory follicular dendritic cells, and thereby closely resembled the B cell follicles seen in lymphoid organs. The T cells were predominantly T helper/inducer cells and in all patients the T cells expressed HLA-DR. One patient diagnosed as lymphocytoma cutis differed from the other patients by having no detectable B cells. One patient with Jessner's lymphocytic infiltrate differed from the other patients by having a marked relative predominance of T suppressor/cytotoxic cells. These data suggest that cutaneous lymphocytoma and lymphoplasia are basically similar disorders which may be considered to be exaggerated immune responses, whereas Jessner's lymphocytic infiltrate may be a separate entity. Immunological analysis may assist in establishing a definite diagnosis in cases of lymphocytoma or lymphoplasia with atypical cytological features.     

Identification in situ of T lymphocytes in the dermal and epidermal infiltrates of mycosis fungoides

Frozen tissue sections from the lesions of five patients with histologically proven mycosis fungoides were examined using a specific antihuman T-cell antiserum and an indirect peroxidase staining technique. The dermal and epidermal infiltrates were shown to comprise predominantly cells bearing the specific T-cell antigen, HTLA. Two of the patients exhibited characteristic Pautrier microabscesses whose cells were exclusively T lymphocytes.     

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.     

草样肉芽肿浸润细胞免疫组织化学表现型的鉴定

应用各种针对T淋巴细胞、T淋巴细胞亚类、B淋巴细胞、单核细胞以及Ia抗原的单克隆及多克隆抗体,对2例覃样肉芽肿的特异性皮损内浸润细胞,用冰冻切片及间接免疫过氧化酶技术进行鉴定.结果为在浸润区内大部份细胞具有T辅助细胞的表面标志,并且Ia抗原阳性.     

Phenotypic characterization of skin-infiltrating cells in pagetoid reticulosis by monoclonal antibodies

The immunological characterization of the infiltrating cells in a patient with a disseminated form of Pagetoid reticulosis (PR) was carried out-in histological section and cell suspensions--by means of a panel of monoclonal antibodies and classical markers (E-rosette and surface immunoglobulins (SIg]. Most of the infiltrating cells were seen to be mature T-lymphocytes with a suppressor/cytotoxic phenotype (T11+, T3+, T8+, T4-). The results suggest that this variant of PR represents a special histological type of cutaneous T cell lymphoma.     

In situ immunological characterization of the infiltrating cells in positive patch tests

Skin biopsies from positive allergic patch tests were analysed by immunoenzymatic labelling of frozen sections with monoclonal antibodies. In seventeen patients the cellular infiltrate consisted of T cells admixed with Langerhans cells/indeterminate cells, but in two patients there were also many B lymphocytes. The B cells were accompanied by dendritic reticulum cells forming B-cell follicles, indistinguishable from those of normal and hyperplastic lymph nodes. There was no correlation between these two immunohistological staining patterns and the sensitizing antigen, the extent of local reaction or the time from epicutaneous application of allergen to examination (2 to 16 days). The ratio between T-helper and T-suppressor cells varied considerably, and showed no correlation with these variables. In all patients the infiltrating T cells expressed HLA-DR antigen. Transferrin receptors were identified on the infiltrating T cells in biopsies from nine patients. These data indicate activation of T cells in the infiltrate from positive patch tests, and support the functional significance of Langerhans cells in the initiation and maintenance of cutaneous contact allergy. An involvement of B cells and B-cell accessory cells in the pathogenesis of contact allergic reactions is also suggested. The presence of dendritic reticulum cells in skin infiltrates from positive patch tests may reflect a functional implication of the skin in the development of B-cell memory.     

A comparison of the dermal lymphoid infiltrates in discoid lupus erythematosus and Jessner''s lymphocytic infiltrate of the skin using the monoclonal antibody Leu 8

Jessners lymphocytic infiltration of the skin (14 cases) and discoid lupus erythematosus (13 cases) were studied and the lymphoid infiltrates in the dermis were compared in the two conditions, using a standard immunoperoxidase technique. Mouse monoclonal antibodies were used to identify T helper lymphocytes, T suppressor lymphocytes and, using the antibody Leu 8, "immunoregulatory lymphocytes". It was shown that the proportions of Leu 8 positive cells was significantly different in the two conditions. The average percentage of Leu 8 positive lymphocytes in the dermal infiltrate found in the cases of Jessner's was 65% (range 40-80%) whereas the average percentage in the cases of discoid LE was 15% (range 2-30%). This observation is further evidence that Jessner's lymphocytic infiltration and chronic discoid lupus erythematosus should be regarded as separate entities.     

T- and B-cell double lymphoma: immunologic characterization using monoclonal antibodies

The coexistence of two non-Hodgkin lymphomas in one patient is extremely rare. We describe a patient who suffered for 17 years from chronic lymphocytic leukemia of B-cell origin with bone marrow and peripheral blood involvement until he developed clinically typical mycosis fungoides of the skin. The neoplastic lymphatic cells in the skin infiltrates were T-cells of the helper/inducer phenotype and thus differed morphologically and by immunological markers from the neoplastic cells in peripheral blood and bone marrow. The clonal B-cell expansion was controlled well over many years by cytotoxic drug therapy, the cutaneous T-cell lymphoma only with limited success by oral photochemotherapy owing to lack of compliance. The patient died 3 years after the diagnosis of MF from a parallel exacerbation of both diseases. Extensive immunohistochemical studies verified the nature of both lymphomas in vivo and at autopsy.     

Reactivity of monoclonal antibody BE 2 in different stages of mycosis fungoides and in benign dermal infiltrates

Summary The cutaneous infiltrate in mycosis fungoides (MF) is predominantly composed of T4-positive T-cells. Attempts to distinguish the early stages of this condition from benign inflammatory infiltrates using anti-T3, T4 and other T-cell-associated antibodies have hitherto been unsucessful. Recently a monoclonal antibody BE 2 has been described as selectively reacting against leukemic cells in patients with cutaneous T-cell lymphoma. To investigate whether the BE 2 antigen is differentially expressed in different stages of MF and benign dermatoses, thus facilitating diagnosis, especially of early MF, the reactivity of monoclonal antibody BE 2 against cutaneous infiltrates from such conditions was assessed.In the early stages of MF only a small number of reactive cells was present. In benign inflammatory infiltrates, especially in those that clinically and histologically were hardly distinguishable from early MF, BE 2 reactivity was essentially the same as in eczematous-stage MF. Lesions from plaque and tumor stage MF contained large numbers of BE 2-reactive cells. Our results indicate that expression of BE 2 is associated with the stage of a given MF lesion and is essentially identical in early MF and eczematous lesions with a similar histopathological appearance.     

In situ identification of mononuclear cells in cutaneous infiltrates in discoid lupus erythematosus, sarcoidosis and secondary syphilis

The inflammatory mononuclear cell infiltrates observed in lesional skin from patients with discoid lupus erythematosus (DLE), sarcoidosis, and secondary syphilis have been characterized in situ. Immunological markers (human T lymphocyte antigens and receptors for sheep erythrocytes, C3b, C3d and Fc gamma) were studied by using immunofluorescence tests with IgG F (ab')2 preparation of anti-T lymphocyte serum, hemadsorption with tissue sections, and tests with soluble immune complexes of peroxidase. In DLE, T lymphocytes were the dominant cell type. In sarcoidosis, the epithelioid cells, including giant cells, had markers similar to macrophages. The lymphohistiocytic cells consisted mainly of macrophages, some T lymphocytes and a few B lymphocytes. In secondary syphilis (condylomata lata), macrophages and T lymphocytes were the dominant cell types, and relatively few B lymphocytes were detected.     

Circulating K and NK cells in malignant melanoma using monoclonal antibodies

K-Cells and NK-Cells are specialized lymphocytes eliciting antibody-dependent cellular cytotoxicity (ADCC) which may play an important part in the immunologic surveillance of various tumors. On the other hand, their cytotoxic impact on neoplastic tissue may be reduced by the tumor itself. By means of monoclonal antibodies (anti-leu 7 = HNK 1), we determined the number of circulating killer cells (K-cells) and natural killer cells (NK-cells) in the peripheral blood of 35 patients with malignant melanoma (MM). Our studies did not reveal any differences in the amount of circulating K-cells or NK-cells between patients with MM at various clinical stages (n = 35) and a control group of healthy persons (n = 29); neither could there be detected any relation towards the age of the patients in both groups. In the advanced clinical stages III and IV of MM (n = 7), however, the number of circulating leu 7-positive cells was obviously increased. Statistically significant (p less than 0.01) was the increase in MM-pT (greater than 3 mm), compared to MM-pT.     

Immunohistochemical study of lepromin reaction in healthy adults

Lepromin reaction was studied with immunoperoxidase techniques using monoclonal antibodies. The skin reactions were induced by injecting Mitsuda antigen into healthy adults without a family history of leprosy. Both early (48 hours) and late (3 weeks and 2 months) reactions were examined. In the late reaction, focal collections of epithelioid cells had formed, and not only T- but also B-lymphocytes were observed around the granuloma. CD1a+ cells were also confirmed to have increased in the late reaction.     

Acute disseminated histiocytosis-X: in situ immunophenotyping with monoclonal antibodies

The proliferating skin cells in a case of acute disseminated histiocytosis-X (Abt-Letterer-Siwe disease) confirmed by electron microscopy, were characterized by a panel of monoclonal antibodies using an immunoperoxidase technique. The "histiocytes" were found to stain with OKT-6 (anti-T6) and anti-HLA-DR antibodies. Unexpectedly, slight staining was also observed with Leu 3a (anti-T4) and OKM-1. A proliferative process of T4, T6, HLA-DR, OKM-1 positive Langerhans' cells has not yet been described and may be specific for histiocytosis-X.