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.     

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.     

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.     

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.     

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.     

Lymphomatoid Papulosis in an 11-month-old Infant

Lymphomatoid papulosis was seen in an 11-month-old child. The condition resolved spontaneously after a course of only 8 weeks and the patient has now been disease free for 9 months. Electron microscopy showed infiltrating lymphocytes with cleaved nuclei suggestive of T cells. Monoclonal antibody studies confirmed the T cell nature of the infiltrate. In this case, suppressor (OKT8) T cells were more prominent than helper (OKT4) T cells, in contrast to previous reports.     

Immunopathologic Demonstration of T Lymphocyte Subpopulations and Interleukin 2 in Graneloma Annulare

Immunopathologic aspects of granuloma annulare were studied in frozen sections of nine skin biopsy specimens with monoclonal antibodies directed against T lymphocytes, Langerhans' cells, interleukin 2, and interleukin 2 receptors in conjunction with immunoperoxidase techniques. The predominant lymphocyte was an activated T lymphocyte (Leu 1+, HLA-DR+) with an excess of helper/inducer phenotype (Leu 3a+) as compared with suppressor/cytotoxic phenotype (Leu 2a+). Langerhans' cells were increased in the epidermis and numerous OKT6+ cells were observed in the perivascular and granulomatous infiltrate. Both interleukin 2-positive cells and interleukin 2 receptor-positive cells were identified in the dermal lesions according to observed reactivity with the corresponding monoclonal antibodies. These findings suggest that a cell-mediated immune response producing cytokines may be important in the pathogenesis of granuloma annulare. Comparison of these results with skin specimens from patients with sarcoidosis and from a patient with granuloma annulare having some of the histologic features of sarcoidosis, suggests that the cutaneous infiltrate in granuloma annulare represents a response distinct from that of sarcoidosis.     

The T cell phenotypes in the lesion of patients with cutaneous leishmaniasis

Immunohistological analysis of the lesions of patients with cutaneous leishmaniasis showed that the OKT8 positive/cytotoxic T suppressor lymphocyte was the predominant cell in the mononuclear cell infiltrate. These patients were nor immunocompromised, had normal cellular immune functions and had developed specific cellular immunity to the infecting parasite, Leishmania tropica major (L. major). These findings support the contention that in patients with cutaneous leishmaniasis inappropriate sensitization of T suppressor lymphocytes occurs, which may, by inhibiting the positive inducer signals of T helper Lymphocytes, account for the chronicity of these lesions.     

An investigation of circulating and in situ lymphocyte subsets and Langerhans cells in the skin and cervix of patients with chronic renal failure

This study was designed to detect possible changes in the immunocytology of the human immune system in the skin, cervix and peripheral blood of patients with chronic renal failure (CRF) treated by conservative methods, haemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD). In the skin, Langerhans cell numbers were reduced in CRF, CAPD and HD patients but in the cervix, Langerhans cells were reduced only in the CRF patients. There was a preponderance of T suppressor lymphocytes compared with T helper lymphocytes in the epidermis in the CRF and the CAPD groups. The presence of natural killer cells in the epidermis of the renal groups compared to controls was significant in the CRF and HD patients while the presence of T suppressor lymphocytes in the epidermis compared to controls was significant only in the CAPD patients. In the dermis, there was a mixed cellular infiltrate of T helper and T suppressor lymphocytes but with no subset attaining significance. The dermal infiltrate of T helper lymphocytes in all three groups of renal patients was significantly reduced compared to controls. In CRF patients, peripheral blood pan T cells, T helper and T suppressor lymphocytes and B lymphocytes were reduced, while T suppressor lymphocytes were reduced in both HD and CAPD patients, compared to controls. Though the results confirm alteration of the immunocytology of the skin, cervix and peripheral blood, we could not relate them to a clinical finding.     

Inflammatory cell types in normal human epidermis--an immunohistochemical and morphometric study

Inflammatory cells in 15 specimens of normal human epidermis were selectively stained by a monoclonal antibody immunoperoxidase technique. The quantitative assessment using an interactive image analysis system revealed OKT 11 positive cells (T lymphocytes) and Leu 2a positive cells (suppressor/cytotoxic cells). As there was no significant difference in the distribution of these markers, helper/inducer cells obviously are not present in considerable amounts. OKM 5 positive cells outnumbered OKM 1 positive cells, indicating the presence of a OKM 5+, OKM 1-macrophage subset. The epidermal dendritic cells clearly showed a striking heterogeneity regarding the expression of HLA-DR (62% of OKT 6-positive cells) and Leu 3a (47%), suggesting the existence of immunologically distinct subsets of human epidermal dendritic cells.     

Immunohistologic studies of squamous cell carcinoma: possible participation of Leu-7+ (natural killer) cells as antitumor effector cells

Immunohistologic studies of 8 patients with squamous cell carcinoma (SCC) were undertaken using a series of monoclonal antibodies. In all of the patients, over 70% of the dermal infiltrates reacted with OKT3 and OKIal (HLA-DR), with a slight predominance of the OKT8+ suppressor/cytotoxic T subset (the mean OKT4/OKT8 ratio was 0.85). Both OKT4+ and OKT8+ subsets could be seen in contact with individual cancer cells. The percentage of OKB7+ (B) cells was less than 29% of the dermal infiltrates. Some Leu-7+ cells (less than 9% of the infiltrates) were seen in close association with individual cancer cells and none of these cells was present apart from the cancer cells. Few OKT6+ cells were observed in the papillary dermis and these had no relation to cancer cells. In the epidermis, OKT6+ dendritic cells remained within normal proportions. Staining with OKM1 revealed sporadic reactive cells. These results strongly suggest that besides T and B lymphocytes, Leu-7+ (natural killer) cells participate in a significant defense mechanism against SCC proliferation.     

Evaluation of acid α-naphthyl acetate esterase activity as a marker of T lymphocytes in benign and neoplastic cutaneous infiltrates

Activity of the enzyme acid α-naphthyl acetate esterase (ANAE) in T lymphocytes and histiocytes in cutaneous tissue sections of patients with histologically confirmed lichen planus, chronic discoid eczema and mycosis fungoides was compared with results obtained using an in situ immunohistochemical technique employing an antiserum against the human T lymphocyte surface antigen (HTLA). In lichen planus and chronic discoid eczema, most of the cells in the cutaneous infiltrate exhibited both positive ANAE activity and the presence of HTLA. In mycosis fungoides, cells identified as T lymphocytes by the presence of HTLA often showed no ANAE activity. ANAE activity appears to be a useful marker of T lymphocytes in benign cutaneous infiltrates but is less reliable in labelling abnormal T cells.     

Increased “in vivo” lymphocyte blastogenesis in the peripheral blood of patients with atopic dermatitis and allergic contact dermatitis

Summary The spontaneous ³H-thymidine (³HT) labelling of some lymphocyte subpopulations has been studied in the peripheral blood of five patients with atopic dermatitis and five with widespread allergic contact dermatitis and compared with that in 10 healthy subjects. One hour after addition of ³HT to heparinized blood, lymphocytes were separated and processed with two different rosetting techniques (E-rosette test and Active-E-rosette test). The cell suspensions were cytocentrifuged and autoradiography undertaken.An increased number of ³HT labelled lymphocytes was observed in the peripheral blood of patients with dermatitis as compared to controls. These labelled lymphocytes were E-rosette-forming cells (T cells) and E-non-rosetteforming cells (non-rosette-forming T cells and non-T cells). The ratio between the labelling index (LI) of E-rosette- to the LI of non-E-rosette-forming cells was in favour of T cells in allergic contact dermatitis (ratio=3.09) whereas in atopic dermatitis (ratio=0.93) the DNA synthesis was relatively greater in the non-rosette-forming cells.It is suggested that this increased LI of peripheral blood lymphocytes could be related to the increased dermal mononuclear cell ³HT-labelling that has been reported previously in these inflammatory skin diseases.D. Van Neste is recipient of a grant from INSERM, Paris and Administration des relations culturelles internationales (ARCI), Bruxelles     

Actinic reticuloid: immunohistochemical analysis of the cutaneous infiltrate in 13 patients

An immunohistological study on cryostat sections of skin biopsies in 13 patients with actinic reticuloid has been performed using a panel of 21 monoclonal antibodies against lymphoid and non-lymphoid infiltrate cells. The infiltrate consisted of activated T cells, numerous histiocytes and macrophages, and small numbers of B cells. In 10 out of 13 patients the majority of the lymphocytes expressed the phenotype of suppressor cells. The number of Leu 8+ cells was inversely proportional to HLA-DR expression by the dermal infiltrate, which suggested a negative correlation between a state of activation and the concentration of Leu 8+ cells. There was a striking number of IgE bearing dendritic cells in the dermis associated with elevated serum IgE levels.     

Expression of human lymphocyte antigen (HLA)-DR on tumor cells in basal cell carcinoma

Immunohistologic studies of eight patients with basal cell carcinoma were undertaken using a series of monoclonal antibodies. In all of the patients, the majority of dermal infiltrates reacted with OKT3 and OKIa1 (HLA-DR), with a slight predominance of OKT4+ helper/inducer T cells (the mean OKT4/OKT8 ratio was 1.8). Both OKT4+ and OKT8+ cells were seen infiltrating the tumor masses. In addition, in five cases, human lymphocyte antigen (HLA)-DR was demonstrated on some tumor cells close to a vast number of HLA-DR+ infiltrates surrounding the carcinoma, but not on epidermal keratinocytes and tumor cells devoid of the HLA-DR+ infiltrates. A considerable number of OKT6+ dendritic cells were also observed surrounding the carcinoma. Staining with OKB7 and OKM1 revealed negligible reactive cells, and virtually none of the dermal infiltrates reacted with Leu-7 (HNK-1). These findings suggest that in addition to varied immunologically competent cells, expression of HLA-DR antigen on tumor cells may participate in a cellular immune reaction, a defense mechanism against tumor cell proliferation in basal cell carcinoma.     

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.     

The localisation of treponemes and characterisation of the inflammatory infiltrate in skin biopsies from patients with primary or secondary syphilis, or early infectious yaws.

OBJECTIVE--To study the localisation of treponemes and to analyse the inflammatory infiltrate in biopsy specimens from patients with primary or secondary syphilis, or early infectious yaws. MATERIALS AND METHODS--Skin biopsies originating from human lesions of primary (29x) or secondary (15x) syphilis (Rotterdam), or early yaws (18x) (West Sumatra) were studied. Different histochemical and immunohistochemical detection methods were used in this study. RESULTS AND CONCLUSION--The histochemical silver staining method according to Steiner revealed the presence of T. pallidum in all cases of primary syphilis studied. In 10 out of 14 cases of secondary syphilis, treponemes were demonstrated. With an immunofluorescence staining technique (IF) using anti-T. pallidum antiserum raised in rabbits (a-Tp), T. pallidum was demonstrated in 28 out of 29 cases of primary syphilis, and in 14 out of 14 studied cases of secondary syphilis. The silver staining method and IF showed identical localisations of T. pallidum (mainly in the dermal-epidermal junction zone or throughout the dermis). Using a-Tp antiserum in the indirect immunofluorescence technique, T. pertenue could be demonstrated in the dermis more often than with Steiner silver staining. However, epidermotropism of T. pertenue in yaws specimens was remarkable, compared with more mesodermotropism of T. pallidum; numbers of T. pertenue in the dermis were limited in all specimens. The dermal inflammatory infiltrate in primary and secondary syphilis was composed mainly of lymphocytes and plasma cells. In most cases more T (CD3 positive) cells than B (CD22 positive) cells were present. Regarding T cell subpopulations, in primary syphilis, T helper/inducer (CD4 positive) cells predominated in 86% of cases. In secondary syphilitic lesions, numbers of T helper/inducer cells were less frequent than or equal to T-suppressor/cytotoxic (CD8 positive) cells in 60% of cases. Remarkably, in yaws specimens the inflammatory infiltrate consisted mainly of IgG, but also IgA and IgM producing plasma cells. T or B lymphocytes were scarce, which is in sharp contrast with findings in syphilitic lesions.     

The presence of IgE molecules on epidermal langerhans cells in patients with atopic dermatitis

Summary Skin sections of clinically involved and clinically normal-looking skin from patients with atopic dermatitis were incubated with anti-human IgE antibodies using the indirect immunoperoxidase technique. Apart from positive dermal anti-IgE staining, positive epidermal anti-IgE staining was also observed. The morphology of the epidermal staining cells suggested the involvement of dendritic cells. This was confirmed by positive immuno-double labelling with OKT6 and anti-IgE. This phenomenon seemed to be specific for atopic dermatitis since skin sections from normal nonatopic controls, patients with allergic asthma, contact dermatitis, and schistosomiasis showed no epidermal anti-IgE staining. To further elucidate the nature of the epidermal anti-IgE staining cells, epidermal cell suspensions were prepared from clinically involved skin from patients with atopic dermatitis. These cell suspensions also showed positive anti-IgE staining cells and positive immuno-double labelling with OKT6 and anti-IgE. Immunogold electron microscopy with anti-IgE on epidermal cell suspensions from patients with atopic dermatitis showed gold particles on the cell membranes of cells containing Birbeck granules, being Langerhans' cells. Epidermal cell suspensions from normal non-atopic controls were negative. The presence of IgE molecules on epidermal Langerhans' cells, which seems to be specific for patients with atopic dermatitis, provides an explanation for the high frequency of positive patch test reactions to inhalant allergens.     

An immunopathological study of herpes-associated erythema multiforme

Any pathogenetic mechanism proposed for erythema multiforme (EM) must account for the prominent mononuclear cell infiltrate in the skin lesions. The purpose of this study was to characterize immunopathologically, with monoclonal antibodies to human leukocyte antigens, the inflammatory cells in early target lesions of recurrent herpes-associated EM. Cryostat sections of snap-frozen skin biopsies were studied by the avidin-biotin immunoperoxidase technique with use of the following monoclonal antibodies: anti-HLA-DR, anti-Leu M5, anti-Leu 4 + 5b, anti-Leu 3a + 3b, anti-Leu 2a, anti-Leu 14, and anti-Leu 6. The dermal mononuclear inflammatory infiltrate in the EM biopsies consisted of monocyte-macrophages and T-lymphocytes, with both helper and suppressor T cells present. Both the dermal inflammatory infiltrate and the overlying keratinocytes were strongly HLA-DR positive. No definite alteration of Langerhans cell number or distribution was noted. These findings are consistent with the characteristics seen in cell-mediated immune reactions in the skin and point to this as a likely immune mechanism for the tissue damage of EM.     

Spitz痣和恶性黑素瘤浸润细胞的免疫细胞化学研究

用多种单克隆抗体,借助间接免疫过氧化物酶技术对1例Spitz痣和2例恶性黑素瘤(MM)Ⅱ, Ⅲ期的皮损进行了观察.结果见在Spitz痣与MM的炎性细胞浸润中,T淋巴细胞(OKT11⁺)占相当比例.在Spitz痣中,T_H/T_S(Leu3a⁺Leu2a⁺)的值与正常人皮肤中的比值相近,而MM的T_H/T_S比值降低.在Ⅳ期MM中,见炎性细胞减少的同时T淋巴细胞的比例下降,并且单核细胞(Monol⁺)数量较多,OKT6⁺细胞缺如.上述结果说明,Spitz痣与MM的炎性浸润细胞在免疫组织化学表现型方面有一定差别,并提示了在MM中细胞免疫功能的异常.