Takatoshi Shimauchi

Augmented expression of programmed death‐1 in both neoplastic and non‐neoplastic CD4+ T‐cells in adult T‐cell leukemia/lymphoma

Adult T‐cell leukemia/lymphoma (ATL) is a CD4+CD25+ T‐cell malignancy infected with human T‐cell leukemia virus type‐I (HTLV‐I). HTLV‐I infection causes the T‐cell dysfunction, which contributes to the immunodeficient state of the patients. Programmed death‐1 (PD‐1) can negatively regulate T‐cell response, when its ligand, PD‐L1 or PD‐L2 mainly expressed on antigen presenting cells, binds to this B7 family receptor. We investigated whether PD‐1 is expressed on CD4+ neoplastic (and/or non‐neoplastic) cells or CD8+ cytotoxic cells in peripheral blood mononuclear cells from 11 patients with ATL. By flow cytometry, we found that the levels of PD‐1 expression on both CD4+CD25+ and CD4+CD25− T‐cell populations were increased in ATL patients compared to normal healthy volunteers, while PD‐1 levels on CD8+ T‐cells were comparable between the patients and normal subjects. In stimulation with anti‐CD3 antibody, the proliferation of PD‐1‐expressing T‐cells from ATL patients was weak when compared to that of PD‐1‐nonexpressing normal T‐cells. In addition to PD‐1, PD‐L1 was coexpressed on ATL cells in some patients, and PD‐L1 expression was enhanced by stimulation with anti‐CD3 antibody. Finally, the production of cytokines such as TNF‐α by ATL cells was restored by blockade of PD‐1/PD‐L1 interaction. These findings suggest that CD4+ T‐cells are the main PD‐1‐expressing cells rather than CD8+ T‐cells in ATL patients, and both neoplastic and normal CD4+ cells are exhausted as a result of PD‐1 expression, and additionally PD‐L1 expression on the neoplastic cell.

Type of skin eruption is an independent prognostic indicator for adult T-cell leukemia/lymphoma

Cutaneous involvement is seen in ~ 50% of adult T-cell leukemia/lymphoma (ATLL) patients. We investigated the association between skin eruption type and prognosis in 119 ATLL patients. ATLL eruptions were categorized into patch (6.7%), plaque (26.9%), multipapular (19.3%), nodulotumoral (38.7%), erythrodermic (4.2%), and purpuric (4.2%) types. When the T stage of the tumor-node-metastasis-blood (TNMB) classification of mycosis fungoides/Sézary syndrome was applied to ATLL staging, 16.0% were T1, 17.7% T2, 38.7% T3, and 4.2% T4, and the remaining 23.5% were of the multipapular and purpuric types. For the patch type, the mean survival time (median survival time could not be estimated) was 188.4 months. The median survival times (in months) for the remaining types were as follows: plaque, 114.9; multipapular, 17.3; nodulotumoral, 17.3; erythrodermic, 3.0; and purpuric, 4.4. Kaplan-Meier curves of overall survival showed that the erythrodermic type had the poorest prognosis, followed by the nodulotumoral and multipapular types. The patch and plaque types were associated with better survival rates. Multivariate analysis demonstrated that the hazard ratios of the erythrodermic and nodulotumoral types were significantly higher than that of the patch type, and that the eruption type is an independent prognostic factor for ATLL. The overall survival was worse as the T stage became more advanced: the multipapular type and T2 were comparable, and the purpuric type had a significantly poorer prognosis than T1.

IL-10-Producing Langerhans Cells and Regulatory T Cells Are Responsible for Depressed Contact Hypersensitivity in Grafted Skin

Although skin grafting is a common surgical technique, the immunological state of grafted skin remains unelucidated. An experimental model has shown that the development of murine contact hypersensitivity (CHS) is depressed when mice are sensitized with a hapten through full-thickness grafted skin. We explored the immunological mechanisms underlying this hyposensitization, focusing on the fate of Langerhans cells (LCs). When FITC was applied to grafted skin, FITC-bearing LCs were capable of migrating to the draining lymph nodes. Epidermal cell suspensions isolated from the grafted skin produced a high amount of IL-10 as assessed by real-time PCR. Adoptive transfer of immune lymph node cells from the sensitized mice suppressed the CHS response of recipients in an antigen-specific manner. CD4(+)CD25(+) but not CD4(+)CD25(-) T cells purified from lymph node cells were responsible for this suppression. Finally, we detected high expression of receptor activators of nuclear factor kappa-B ligand (RANKL) in the grafted skin, and found that recombinant RANKL stimulated LCs to produce IL-10. These findings suggest that the hyposensitization of CHS through the grafted skin is not attributable merely to the reduction of LC number but that IL-10-producing LCs exert a downmodulatory effect by inducing regulatory T cells.

Aberrant aquaporin 5 expression in the sweat gland in aquagenic wrinkling of the palms

Aquagenic wrinkling of the palms (AWP) is an uncommon disease characterized by the rapid and transient formation of edematous whitish plaques on the palms on exposure to water. Although this disease is occasionally accompanied by hyperhidrosis, the pathophysiology of AWP remains unknown. Herein we describe a patient with AWP. The location of wrinkling was limited to the areas positive for iodine-starch test after water exposure, which suggests that AWP is etiologically related to hyperhidrosis. Histologic examination revealed hyperplastic and papillated eccrine glandular epithelium with the enlarged diameter of eccrine coils. Immunohistochemically, while aquaporin 5 (AQP5), one of the water channel AQP families, was present exclusively in the dark cells of sweat glands of healthy donors, an aberrant AQP5 staining, extending to the clear cells, was found in the patient with AWP. The hyperplastic glandular epithelium and aberrant AQP5 staining in the patients sweat glands suggest that AWP stems from dysregulation of sweating.

Adult T-cell leukemia/lymphoma cells from blood and skin tumors express cytotoxic T lymphocyte-associated antigen-4 and Foxp3 but lack suppressor activity toward autologous CD8+ T cells

Adult T cell leukemia/lymphoma (ATL) cells share the CD4+CD25+ phenotype with regulatory T (Treg) cells. However, it is still controversial whether ATL cells are Treg cells. The aim of the present study was to investigate the Treg nature of ATL cells obtained from peripheral blood and skin tumors in terms of their phenotype and function. By flow cytometry and immunohistochemistry, the expression of the Treg‐associated molecule cytotoxic T lymphocyte‐associated antigen (CTLA)‐4 and Foxp3 was examined in freshly isolated circulating and skin‐infiltrating tumor cells from 21 ATL patients with skin eruptions. The expression of CTLA‐4 on freshly isolated circulating tumor cells was elevated in two of 15 patients, and Foxp3 was expressed intracytoplasmically at high levels in three of nine patients. In five of the patients examined, skin‐infiltrating tumor cells bore variously elevated CTLA‐4 with high Foxp3 expression. The potentiality of ATL cells as Treg cells was further addressed by stimulating ATL cells with anti‐CD3/CD28 monoclonal antibodies and monitoring CTLA‐4 expression. With the stimulation, even CTLA‐4‐low ATL cells expressed higher levels of CTLA‐4 than normal CD4+CD25+ cells. To study function, ATL cells isolated from blood and skin tumors were tested for their ability to suppress the proliferation of autologous CD8+ T cells stimulated with allogeneic lymphocytes. Despite the expression of CTLA‐4 and Foxp3, these tumors were incapable of suppressing the proliferation of autologous CD8+ T cells. ATL cells are phenotypically Treg cells in at least some patients, but lack immunoregulatory functions, at least toward CD8+ T cells. (Cancer Sci 2008; 99: 98–106)

Production of Thymus and Activation-Regulated Chemokine and Macrophage-Derived Chemokine by CCR4+ Adult T-Cell Leukemia Cells

Purpose: Adult T-cell leukemia/lymphoma (ATL) is a peripheral CD4+CD25+ T-cell malignancy caused by human T-cell leukemia virus type I. The tumor cells frequently infiltrate in the skin, lymph nodes and other organs and especially form prominent cutaneous masses. Recently, ATL cells have been shown to express Th2 chemokine receptor CCR4. The aim of this study is to investigate the possibility that CCR4 ligands, thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC), are produced by CCR4+ ATL cells per se. Experimental Design: CD4+ or CD4+CD14− cells were purified from peripheral blood mononuclear cells of 11 ATL patients with cutaneous involvement and normal healthy volunteers. Tissue-infiltrating cells were isolated from skin tumors. The expression of chemokine receptors on these cells were analyzed by flow cytometry. The production of chemokines and cytokines by the neoplastic cells was assessed by ELISA and reverse transcription-PCR after cultivation for 96 hours in the presence or absence of anti-CD3/CD28 monoclonal antibodies. Finally, TARC and CCR4 expressions were examined by immunohistochemistry. Results: ATL cells highly expressed CCR4 but did not necessarily exhibit the Th2 cytokine profile. The cells also produced TARC and MDC. The production level of MDC was higher in the skin tumor formation group than that in the nontumor group. Immunohistochemically, both CCR4 and TARC were expressed by the tumor cells in the lesional skin. Conclusions: ATL cells not only express CCR4 but also produce TARC and MDC. The skin tumor formation as well as the monoclonal integration of proviral DNA are the factors that are associated with the high production of Th2 chemokines by ATL cells.

Inhibition of T helper 2 chemokine production by narrowband ultraviolet B in cultured keratinocytes

Background  Narrowband ultraviolet B (NB‐UVB) has recently been used for the treatment of various skin disorders. Its effects on the production of cytokines and chemokines by keratinocytes are unknown.

Alterations of serum Th1 and Th2 chemokines by combination therapy of interferon-γ and narrowband UVB in patients with mycosis fungoides

BACKGROUND Mycosis fungoides (MF) is a T cell neoplasm with elevation of serum Th2 chemokines. Although interferon-gamma (IFN-gamma) administration and narrowband-UVB (NB-UVB) phototherapy are used for the treatment of MF, a combination therapy of these two modalities is not fully established. OBJECTIVES To define whether the combination of IFN-gamma and NB-UVB affects the balance of serum levels of Th1 and Th2 chemokines in patients with MF. METHODS Twelve patients with MF received intravenous or intramuscular injections of recombinant IFN-gamma (rIFN-gamma) or natural IFN-gamma (nIFN-gamma) in combination with NB-UVB phototherapy. As control, three MF patients were treated with NB-UVB monotherapy. At the beginning and cessation of therapy, the concentrations of serum Th2 chemokines, TARC/CCL17 and MDC/CCL22, and Th1 chemokines, IP-10/CXCL10 and MIG/CXCL9 were measured by ELISA. RESULTS Before treatment, not only Th2 chemokines but also Th1 chemokines were elevated in the patients. Whereas no significant changes were observed in the levels of TARC and MDC, IP-10 and MIG were further elevated by the combination of IFN-gamma and NB-UVB. On the other hand, NB-UVB monotherapy did not change the level of either Th1 or Th2 chemokine. CONCLUSIONS The combination of IFN-gamma and NB-UVB elevated serum Th1 chemokines but unaffected Th2 chemokines. Since NB-UVB monotherapy could not affect the chemokine levels, the effect of the combination therapy is attributable to IFN-gamma. Given the role of Th1 chemokines for tumor-attacking T cell recruitment at the early stage of MF, the therapy may exert a beneficial effect for early MF.

Defective epidermal innate immunity and resultant superficial dermatophytosis in adult T cell leukemia/lymphoma

Purpose: Superficial dermatophytosis is quite commonly seen in patients with adult T-cell leukemia/lymphoma (ATLL), as approximately 50% of the patients develop cutaneous mycotic infections. Because superficially infected fungi in the stratum corneum of the epidermis cannot directly contact with T cells infiltrating in the upper dermis, some perturbation of epidermal innate immunity has been postulated. Interleukin (IL)-17–producing helper T cells (Th17) can induce the keratinocyte production of antimicrobial peptides such as human β defensin (HBD)-2 and LL-37, which play an essential role in cutaneous innate immunity. Experimental Design: We investigated the frequency of circulating Th17 cells, serum levels of cytokines, and epidermal expression of HBD-1, 2, 3, and LL-37 in ATLL patients with or without superficial dermatophytosis. Results: The frequency of peripheral Th17 cells and the serum level of IL-17 was significantly decreased in ATLL patients, whereas the serum IL-10 and TGF-β1 levels were increased as compared with healthy controls. Furthermore, ATLL patients with dermatophytosis had higher IL-10 and TGF-β1 levels and lower IL-17 levels than did those without dermatophytosis. Immunohistochemical study revealed that the epidermal expression of both HBD-2 and LL-37 were significantly lower in ATLL patients with dermatophytosis than in non-ATLL patients with dermatophytosis. Conclusions: Taken together, these results suggest that the keratinocyte production of antimicrobial peptides promoted by Th17 cells is reduced in ATLL patients, leading to the perturbed innate immunity and the frequent occurrence of superficial dermatophytosis. Clin Cancer Res; 18(14); 3772–9. ©2012 AACR.

Purpuric adult T-cell leukaemia/lymphoma: expansion of unusual CD4/CD8 double-negative malignant T cells expressing CCR4 but bearing the cytotoxic molecule granzyme B

A 78‐year‐old Japanese woman with adult T‐cell leukaemia/lymphoma (ATL) presented with an unusual purpuric and erythematous eruption on the face and trunk. Immunohistochemical and flow cytometric analyses showed that the tumour cells were CD4/CD8 double‐negative, and expressed CCR4 T‐helper (Th) 2 chemokine receptors. Despite these features, the cells aberrantly produced granzyme B, which is a cytotoxic molecule usually produced by CD8+ cytotoxic T cells, natural killer cells, or occasionally by Th1 cells. In a purpuric lesion, extravasation of erythrocytes was associated with an infiltrate of these cytotoxic tumour cells. Our case suggests phenotypical and functional heterogeneity of tumour cells in ATL, which may be closely related to the clinical appearance of the skin eruption.