Immunological markers of Langerhans' cells in mycosis fungoides skin lesions

The patterns of two immunological markers of epidermal Langerhans' cells were compared in normal skin and mycosis fungoides (MF) skin lesions. A double-staining indirect immunofluorescence method was used. Cells suggestive of keratinocytes reacted with the anti-Ia antibodies in some cases of MF lesions, while no such pattern was seen with the thymocyte-antigen antibodies (OK T6). In other cases of MF, as in normal skin, the epidermal picture was mainly the same with the two markers. A great many of the dermal mononuclear cells in MF were reactive with the anti-Ia antibodies but only occasional dermal cells in MF and no dermal cells in normal skin stained with OK T6.     

Blood lymphocyte subpopulations in Mycosis fungoides and their functions in vitro

T-lymphocyte subpopulations, PWM stimulated in vitro Ig production by mononuclear cells, and spontaneous cell-mediated cytotoxicity (SCMC) were determined in patients with Mycosis fungoides (MF) as well as in sex- and age-matched normal controls. We were able to confirm our earlier findings of a significantly higher frequency of T-lymphocytes with Fc-receptors for IgG (Tg cells) and a proportion of T lymphocytes with Fe receptors for IgM (Tm cells) not significantly differing from normal controls. As defined by monoclonal antibodies, the proportion of T-helper cells (Leu3A) was significantly lower, whereas the T-suppressors cells (Leu2A) were in the range of normal controls. The PWM stimulated and spontaneous Ig synthesis in vitro was lower in MF patients whereas there was no difference in the SCMC activity, in comparison with normal controls. The lower frequency of T-helper cells in the peripheral blood may be explained by their migration to the skin.     

A comparative immunohistochemical study of adult T cell leukemia and cutaneous T cell lymphoma

An immunoperoxidase study of 2 cases of adult T cell leukemia (ATL) and 2 cases of mycosis fungoides (MF) was done using monoclonal antibodies. In ATL, many anti-interleukin-2 receptor antibody (LeuIL-2R)-positive cells were seen in the dermis and occasionally in the epidermis. In contrast, in MF, LeuIL-2R-positive cells were much less frequent. LeuIL-2R-positive cells in MF may be non-malignant T cells; not all LeuIL-2R-positive cells may be malignant in ATL. These non-malignant LeuIL-2R-positive cells, we suggest, are involved in the interaction between malignant T cells and reactive infiltrating cells. Furthermore, in addition to OKT6-positive cells, OKM1-positive cells were seen in the infiltrates in the dermis in both ATL and MF. OKM1-positive cells also participate in the mechanism of the skin affinity in ATL and cutaneous T cell lymphomas.     

Expression of Fas and Fas-ligand in primary cutaneous T-cell lymphoma (CTCL): association between lack of Fas expression and aggressive types of CTCL

BACKGROUND: Fas (CD95; APO-1) is a transmembrane protein that mediates apoptosis upon cross-linking with Fas-ligand (Fas-L). Interaction of Fas-L expressed by cytotoxic T cells with Fas-expressing tumour cells plays an important part in antitumour immune responses. OBJECTIVES: We aimed to investigate Fas and Fas-L expression in frozen and paraffin-embedded material from a large group of patients with cutaneous T-cell lymphoma (CTCL). METHODS: Immunostaining with monoclonal antibodies against Fas and Fas-L was performed in material from 23 patients with mycosis fungoides (MF), 10 with lymphomatoid papulosis (LyP), 10 with CD30-positive primary cutaneous large T-cell lymphoma (LTCL) and nine with CD30-negative LTCL. The results were correlated with the type and stage of CTCL and clinical features. RESULTS: Expression of Fas by the large majority of the neoplastic T cells was observed in 15 of 15 cases of plaque-stage MF, 10 of 10 cases of LyP and 10 of 10 cases of CD30-positive LTCL, but only in four of 12 cases of tumour-stage MF and two of nine cases of CD30-negative LTCL. In three of four MF patients in whom both plaques and tumours could be studied, a significant decrease in Fas expression was observed with progression from plaque-stage to tumour-stage disease. Fas-L was expressed by > 50% of the neoplastic T cells in 46 of 56 biopsies, and no clear relationship with type of CTCL and clinical behaviour was observed. CONCLUSIONS: This study demonstrates loss of Fas expression in aggressive types of CTCL, but not in indolent types of CTCL. These data suggest that loss of Fas receptor expression may be one of the mechanisms that allow tumour cells to escape an effective immune response, and may contribute to the unfavourable prognosis of some types of CTCL.     

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.     

Decreased interleukin‐21 expression in skin and blood in advanced mycosis fungoides

Interleukin (IL)‐21 is regarded as a potent antitumor agent, which increases the cytotoxicity of both natural killer (NK) and CD8⁺ T cells. In this study, we investigated the role of IL‐21 in mycosis fungoides (MF). IL‐21 mRNA expression levels in patch and plaque MF were significantly higher than those in normal skin. IL‐21 mRNA expression levels in tumor MF were significantly decreased compared with those in patch and plaque MF. Interestingly, mRNA expression levels of IL‐21 in MF lesional skin significantly correlated with those of T‐helper type‐1 cytokines/chemokines such as CXCL10, CXCL11 and γ‐interferon. Immunohistochemistry showed that IL‐21 was expressed by keratinocytes in patch and plaque MF. Furthermore, serum IL‐21 levels in patients with tumor MF were significantly lower than those of healthy controls and plaque MF. Thus, IL‐21 expression was significantly downregulated in skin and blood of patients with tumor MF, which may contribute to progression of MF. Our study suggests that recombinant IL‐21 would be a promising therapy for MF.     

T lymphocyte subpopulations in alopecia areata and psoriasis: identification with monoclonal antibodies and Fc receptors

A comparison of T lymphocyte subpopulations as defined by Fc receptors and monoclonal antibodies was investigated in 9 patients with alopecia areata and alopecia universalis (AA and AU) and in 6 patients with psoriasis. It was shown that there was higher proportion of T lymphocytes with Fc receptors for IgG (Tg cells) in patients with alopecia (AA and AU) and psoriasis. The proportions of total T lymphocytes (Tt), T lymphocytes with Fc receptors for IgM, T suppressor/cytotoxic cells (Leu2A), T helper/inducer (Leu3A) as defined by monoclonal antibodies were within normal range as compared to the normal donors. The possible reason of the dissociation between Tg and T suppressor (Leu2A) cells could be that these cells belong to different subpopulations.     

Treatment of severe psoriasis with low dose cyclosporin A and the effect on the helper-suppressor T cell ratio in peripheral blood

Oral administration of cyclosporin A (CsA) 14 mg/kg/day has been reported to improve psoriasis in a double-blind study. The purpose of this study was to evaluate the efficacy of a lower dose of CsA in severe psoriasis and to monitor lymphocyte subpopulations in peripheral blood in order to detect its mechanism of action. Eleven patients with severe, active psoriasis were treated only with oral administration of 5 mg/kg/day CsA twice a day for 3 months, during which they were closely examined, including single- and two-color analysis of lymphocyte surface markers by flow cytometry using monoclonal antibodies. Seven patients showed improvement within a week, and the others within 2-3 weeks. Five patients had total remission, 2 showed marked improvement, and 4 showed moderate improvement; no clinically important side effects, except hypertrichosis in 2 females were seen. In the peripheral blood of patients treated with CsA there was a decrease in the percentages of helper inducer (CD4⁺4B4⁺)T(Thi) cells, while no significant decrease was found in those of suppressor inducer (CD4⁺2H4⁺)T(Tsi), suppressor effector (CD8⁺CD11⁺)T(Tse), or cytotoxic (CD8⁺CD11^-)T(Tc) cells, resulting in a decrease in the ratio of Thi to Tsi, Tse and Tc. The activated helper (CD4⁺HLA-DR⁺)—suppressor (CD8⁺HLA-DR⁺)T cell ratio was also decreased. These immunological findings obtained from the patients were consistent with in vitro studies of CsA reported earlier and may well explain the effectiveness of CsA in psoriasis as observed in this study.     

Primary cutaneous CD30+ lymphoproliferative disorders

Primary cutaneous CD30⁺ lymphoproliferative disorders are the second most common group of cutaneous T‐cell lymphomas (CTCL) and include lymphomatoid papulosis (LyP) and primary cutaneous anaplastic large T‐cell lymphoma (cALCL). Both disease entities share overlapping clinical, histopathological, and molecular features, thus representing a spectrum of cutaneous CD30⁺ lymphoproliferative disorders. LyP may be distinguished from cALCL by clinicopathological correlation. In some patients, both diseases may coexist at initial diagnosis or develop over the course of the disease. Mycosis fungoides (MF), the most common CTCL, is not considered a primary cutaneous CD30⁺ lymphoproliferative disorder, but may occur in some LyP patients. In addition, LyP‐like lesions may develop in MF patients. However, this is frequently a manifestation of MF rather than a representation of two different disease entities. Caution also has to be taken in the setting of transformed MF with lesions expressing CD30, as they may be mistaken for either LyP or cALCL, resulting in an inadequate therapeutic approach.     

Secukinumab for treatment of psoriasis: does secukinumab precipitate or promote the presentation of cutaneous T‐cell lymphoma?

Secukinumab is an interleukin (IL)‐17 monoclonal antibody inhibiting T‐helper (Th)1‐mediated immune response. It has proven high efficacy for moderate to severe psoriasis but data on its long‐term toxicities are limited. We describe two patients who received secukinumab for clinically presumed psoriasis, but were subsequently diagnosed with mycosis fungoides (MF) following skin biopsies triggered by skin deterioration while on secukinumab. Previous studies suggested decreased numbers of regulatory T cells (Tregs) with increasing stage of MF, which may lead to the shift in the Treg/Th17 balance towards the Th17 pathway. Theoretically, the use of IL‐17 monoclonal antibodies to inhibit Th17 pathway may lead to further immunosuppression and disease progression in cutaneous T‐cell lymphoma (CTCL) by shifting the balance towards Tregs, although this hypothesis has not been proven. With uncertainty over the role of IL‐17 and Treg/Th17 as well as diagnostic challenges in CTCL, we recommend that patients should have a confirmatory skin biopsy prior to the commencement of biologic therapy.     

Spectral karyotyping demonstrates genetically unstable skin-homing T lymphocytes in cutaneous T-cell lymphoma

We initially established cell lines from skin biopsies from four patients (MF8, MF18, MF19 and MF31) in early stages of cutaneous T-cell lymphoma (CTCL) in 1999. After 3 weeks of culture, skin-homing T lymphocytes were stimulated with phytohaemagglutinin. Metaphase spreads were analysed using spectral karyotyping (SKY), a molecular cytogenetic technique. MF18 and MF19 had predominantly normal karyotypes. MF8 had recurrent numerical aberrations resulting in two T lymphocyte clones: one with trisomy 21 (12/20 cells) and the other with monosomy chromosome 22 (3/20 cells). MF8 also exhibited a clonal deletion, del(5)(p15.1), as well as multiple non-clonal structural aberrations. MF31 had a clonal deletion, del(17)(p12) and other non-clonal deletions involving chromosomes 2, 5, 10, 11. MF18 had a single abnormal cell that contained two reciprocal translocations t(1;2)(q32;p21) and t(4;10)(p15.2;q24). In 2001, three of the original patients had new skin biopsies taken and cell lines were established. SKY analysis revealed the continued presence of a T-cell clone in MF8 with trisomy 21 (4/20 cells). Additionally, a new clone was seen with a del(18)(p11.2) (17/20 cells). MF31 had only one aberrant cell with a del(17)(p12). MF18 had a clonal deletion, [del(1)(p36.1) in 3/20 cells] and non-clonal aberrations involving chromosomes 3, 4, 5, 6, 12, 13, 17 and 18. Thus, three of four patients continued to show numerous numerical and structural aberrations, both clonal and non-clonal, with only MF8 having a recurring T lymphocyte clone (+21). Our findings demonstrate high genetic instability among skin-homing T lymphocytes even in early stages of CTCL. We did not see genetic instability or evidence of clones in cell lines from a patient with atopic dermatitis and one with psoriasis.     

Ultraviolet A1 phototherapy for mycosis fungoides

Background. Mycosis fungoides (MF) is the most common form of primary cutaneous lymphoma, and is characterized by a malignant proliferation of CD4+ cells. Psoralen ultraviolet A (PUVA) irradiation is the most common treatment for cutaneous lesions. However, PUVA carries the risk of adverse reactions to psoralens and long‐term risk of skin cancer. UVA1 may be a safer alternative. Aim. To assess the efficacy of UVA1 phototherapy in patients with early‐stage MF (T1–T2). Methods. Four patients with early‐stage MF were treated with 1630–2710 J/cm² UVA1 given in 29–40 fractions, and the effect was assessed by clinical examination and by high‐resolution ultrasonography. Results. Complete clinical remission of MF was achieved in all cases. Conclusions. This preliminary report indicates that UVA1 phototherapy might be an efficient treatment for early‐stage MF.     

Restricted T-cell receptor Vβ gene usage in the skin of patients with guttate and chronic plaque psoriasis

A strong association between acute guttate psoriasis and group A, β-haemolytic streptococcal infections is well established. Furthermore, streptococcal M proteins and toxins have been shown to act as superantigens, stimulating subpopulations of T lymphocytes expressing particular Vβ families. We have therefore studied the possible role of streptococcal superantigens in psoriasis by staining peripheral T lymphocytes and skin sections from patients with guttate or chronic plaque psoriasis for the expression of nine TCR Vβ families, using a range of monoclonal antibodies. A marked over-representation of Vβ2⁺ T lymphocytes was observed in the dermis and epidermis of patients in both groups, when compared with T lymphocytes in their peripheral blood. A less marked dermal increase in Vβ5.1⁺ T lymphocytes was also observed in these patients. These findings are consistent with the involvement of a superantigen, possibly streptococcal, in the pathogenesis of psoriasis.     

The small-cell variant of mycosis fungoides. A clinicopathological and quantitative electron microscopic study on 14 patients

Abstract Small-cell variants of Sézary syndrome and mycosis fungoides (MF) have been described. However, in these studies the nuclear area of the small-cell variant of MF (SC-MF) as compared to histological classical MF (CL-MF) was not characterized objectively by quantitative electron microscopy. In a 14-year follow-up period, of a total of 76 patch/plaque stage MF patients seen in the Department of Dermatology of the University Hospital Utrecht, 14 (18%) had an infiltrate composed of atypical lymphocytes characterized by a distinctly smaller cell diameter and smaller, hyperchromatic, deeply indented nuclei as compared to the usual cell type of MF. The aim of the investigation was to confirm this observation objectively using quantitative electron microscopy (morphometry) and to define SC-MF as compared to CL-MF. The study was performed on the 14 patients with SC-MF, and 10 patients with clinical and histological CL-MF and 4 patients with chronic eczema. Electron micrographs of sections obtained from each biopsy were analysed by computer to produce the following data: a nuclear contour index (NCI), the mean nuclear area (MNA), the mean nuclear area of the cells above the 75th percentile (P75NA) and the percentage of cells larger than 30 μm². The values of MNA differed significantly between patients with SC-MF and those with CL-MF (17.6 vs 23.2 μm²; P = 0.02), as did the values of P75NA (20.7 vs 27.9 μm²; P = 0.01). The NCI of the SC-MF and CL-MF patients were similar. These results are consistent with our observations that SC-MF does indeed exist. Received: 7 April 1998 / Received after revision: 27 July 1998 / Accepted: 7 August 1998     

CCR4 is expressed on infiltrating cells in lesional skin of early mycosis fungoides and atopic dermatitis

CCR4 is expressed on tumor cells of mycosis fungoides (MF) and Sézary syndrome (SS). In MF, most infiltrating cells in patches and plaques express CXCR3, while tumor cells express CCR4 in advanced stages. Poteligeo Test IHC (CCR4 staining kit) is a newly developed staining kit that can examine the presence of CCR4 expressed on tumor cells of adult T‐cell leukemia/lymphoma, peripheral T‐cell lymphoma and cutaneous T‐cell lymphoma before treatment of anti‐CCR4 antibody using paraffin‐embedded samples. In this study, we analyzed CCR4 expression in lesional skin of MF, SS, atopic dermatitis (AD) and psoriasis with this new kit. CCR4 was expressed on infiltrating cells in lesional skin of patch, plaque, tumor MF and SS, and the number of positive cells increased as the disease progressed. Immunohistochemistry with frozen sections also showed some positive cells scattered in the dermis, although the quality was not high enough to quantify positive cells. There were significant positive correlations between CCR4⁺ cells and serum lactate dehydrogenase levels. Interestingly, CCR4⁺ cells were also detected in AD skin, whose number was larger than that in psoriatic skin. Previous studies showed only scattered CCR4⁺ cells in skin samples by standard immunohistochemical staining. The new, sensitive CCR4 staining kit has revealed that CCR4 is expressed on infiltrating cells in lesional skin of early MF and AD as well as advanced MF and SS. These cells can be therapeutic targets for patients who are resistant to standard treatments.     

Expression of complement receptor CR2 (CD21) on human subcorneal keratinocytes in normal and diseased skin.

Human keratinocytes are able to synthesize and express cell surface moieties characteristic of effector and/or accessory cells of the immune system (CD16, CD36, HLA-DR, intercellular adhesion molecule-1). In the present study, skin biopsies from healthy volunteers, from patients with psoriasis vulgaris (PV), mycosis fungoides (MF), purpura pigmentosa chronica (PPC), acute urticaria (AU) and from positive tuberculin skin tests were investigated with regard to the reactivity with the monoclonal antibodies to complement receptors CR1 CR2 and CR3 by means of a multistep immunoperoxidase method. In the clinically involved skin of all patients with PV, MF or PPC, and in biopsies obtained from positive tuberculin tests, specific epidermal intercellular staining with OKB7 and Leu anti-CR2 was seen on subcorneal keratinocytes. This finding suggests a differentiation-linked expression of CR2 on human keratinocytes in cytokine-mediated skin diseases whereas CR1 and CR3 are apparently not expressed.     

Hematopoietic stem cell transplantation in advanced cutaneous T‐cell lymphoma

We retrospectively reviewed data pertaining to five patients with cutaneous T‐cell lymphoma (CTCL) who had received hematopoietic stem cell transplantation (HSCT) between 2004 and 2015 at Kurume University Hospital, along with their clinical data until March 2016. For patients with advanced CTCL eligible for HSCT, autologous HSCT was performed when they responded well to chemotherapy, and allogeneic HSCT was selected for patients with advanced mycosis fungoides (MF)/Sézary syndrome (SS) and CTCL other than MF/SS with poor chemosensitivity. Two patients (primary cutaneous anaplastic large cell lymphoma and primary cutaneous CD8⁺ aggressive epidermotropic cytotoxic T‐cell lymphoma) who responded well to chemotherapy received autologous HSCT: one patient was alive in partial remission and the other died due to therapy‐related acute myeloid leukemia without disease relapse. In the remaining three patients with MF or SS, allogeneic HSCT was performed. Although one patient with MF died due to disease progression, the remaining two patients were alive in complete remission. Although there were two deaths in this study, the outcomes were considered satisfactory.     

Effect of syphilitic rabbit sera taken at different periods after infection on treponemal motility, treponemal attachment to mammalian cells in vitro, and treponemal infection in rabbits.

The time course of antibody synthesis during syphilis was studied in experimentally infected rabbits. A rapid antibody response was seen; the rabbits became positive in both the rapid plasma reagin (RPR) test and Treponema pallidum haemagglutination assay (TPHA) by nine days after infection. Treponemal immobilising antibodies were also seen as early as nine days after infection. Antibody inhibition of treponemal attachment to baby rabbit genital organ (BRGO) cells in culture occurred with immune sera taken 30 days after infection but not earlier. When T pallidum was mixed with immune syphilitic rabbit sera taken at different stages of the infection and used to infect normal rabbits the rabbits became partially resistant to T pallidum only when the treponemes were mixed with sera taken at least 30 days after syphilitic infection. This appearance correlated well with the development of antibodies which blocked attachment of T pallidum to host cells. These antibodies may be involved in the resistance to reinfection which develops in syphilis as the disease progresses.     

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.     

Human leukocyte antigens in cutaneous T cell lymphoma

Mycosis fungoides (MF) and Sézary syndrome (SS) are malignant non-Hodgkin's lymphomas, characterized by the proliferation of helper type T lymphocytes with a predilection for the skin. Because of the similarities in cytologic, histologic, cytogenetic, immunologic, and functional aspects of the malignant cells, as well as overlapping clinical features, these disorders are currently classified as cutaneous T cell lymphoma (CTCL). Though the etiology of these disorders remains obscure, environmental factors as well as viral infection have been implicated. In this study, seventy-six white patients with CTCL were typed for human leukocyte antigen (HLA)-A, -B, and -C to assess genetic susceptibility as determined by the major histocompatibility complex. An increase in the frequency of B8 and Bw35 was seen in SS patients but not in MF patients.