Tomomitsu Miyagaki

Regulatory B cells control T-cell autoimmunity through IL-21-dependent cognate interactions

B cells regulate immune responses by producing antigen-specific antibodies. However, specific B-cell subsets can also negatively regulate T-cell immune responses, and have been termed regulatory B cells. Human and mouse regulatory B cells (B10 cells) with the ability to express the inhibitory cytokine interleukin-10 (IL-10) have been identified. Although rare, B10 cells are potent negative regulators of antigen-specific inflammation and T-cell-dependent autoimmune diseases in mice. How B10-cell IL-10 production and regulation of antigen-specific immune responses are controlled in vivo without inducing systemic immunosuppression is unknown. Using a mouse model for multiple sclerosis, here we show that B10-cell maturation into functional IL-10-secreting effector cells that inhibit in vivo autoimmune disease requires IL-21 and CD40-dependent cognate interactions with T cells. Moreover, the ex vivo provision of CD40 and IL-21 receptor signals can drive B10-cell development and expansion by four-million-fold, and generate B10 effector cells producing IL-10 that markedly inhibit disease symptoms when transferred into mice with established autoimmune disease. The ex vivo expansion and reinfusion of autologous B10 cells may provide a novel and effective in vivo treatment for severe autoimmune diseases that are resistant to current therapies.

Cutaneous Lymphoma International Consortium Study of Outcome in Advanced Stages of Mycosis Fungoides and Sézary Syndrome: Effect of Specific Prognostic Markers on Survival and Development of a Prognostic Model

PURPOSE Advanced-stage mycosis fungoides (MF; stage IIB to IV) and Sézary syndrome (SS) are aggressive lymphomas with a median survival of 1 to 5 years. Clinical management is stage based; however, there is wide range of outcome within stages. Published prognostic studies in MF/SS have been single-center trials. Because of the rarity of MF/SS, only a large collaboration would power a study to identify independent prognostic markers. PATIENTS AND METHODS Literature review identified the following 10 candidate markers: stage, age, sex, cutaneous histologic features of folliculotropism, CD30 positivity, proliferation index, large-cell transformation, WBC/lymphocyte count, serum lactate dehydrogenase, and identical T-cell clone in blood and skin. Data were collected at specialist centers on patients diagnosed with advanced-stage MF/SS from 2007. Each parameter recorded at diagnosis was tested against overall survival (OS). RESULTS Staging data on 1,275 patients with advanced MF/SS from 29 international sites were included for survival analysis. The median OS was 63 months, with 2- and 5-year survival rates of 77% and 52%, respectively. The median OS for patients with stage IIB disease was 68 months, but patients diagnosed with stage III disease had slightly improved survival compared with patients with stage IIB, although patients diagnosed with stage IV disease had significantly worse survival (48 months for stage IVA and 33 months for stage IVB). Of the 10 variables tested, four (stage IV, age > 60 years, large-cell transformation, and increased lactate dehydrogenase) were independent prognostic markers for a worse survival. Combining these four factors in a prognostic index model identified the following three risk groups across stages with significantly different 5-year survival rates: low risk (68%), intermediate risk (44%), and high risk (28%). CONCLUSION To our knowledge, this study includes the largest cohort of patients with advanced-stage MF/SS and identifies markers with independent prognostic value, which, used together in a prognostic index, may be useful to stratify advanced-stage patients.

Eotaxins and CCR3 Interaction Regulates the Th2 Environment of Cutaneous T-Cell Lymphoma

CC chemokine receptor 3 (CCR3), the sole receptor for eotaxins, is expressed on eosinophils and T helper type 2 (Th2) cells. In Hodgkins disease, eotaxin-1 secreted by fibroblasts collects Th2 cells and eosinophils within the tissue. Similarly, many Th2 cells infiltrate the lesional skin of cutaneous T-cell lymphoma (CTCL). In this study, we investigated the role of eotaxins in the development of the Th2 environment of CTCL. We revealed that fibroblasts from lesional skin of CTCL expressed higher amounts of eotaxin-3 messenger RNA (mRNA) compared with those from normal skin. Lesional skin of CTCL at advanced stages contained significantly higher levels of eotaxin-3 and CCR3 mRNA, compared with early stages of CTCL. IL-4 mRNA was expressed in some cases at advanced stages. Immunohistochemistry revealed that keratinocytes, endothelial cells, and dermal fibroblasts in lesional skin of CTCL showed a stronger expression of eotaxin-3 than did normal skin. CCR3(+) lymphocytes and IL-4 expression were observed in some cases of advanced CTCL. Furthermore, both serum eotaxin-3 and eotaxin-1 levels of CTCL patients at advanced stages were significantly higher than those of healthy individuals. The concentrations of these chemokines correlated with serum soluble IL-2 receptor levels. These results suggest that interaction of eotaxins and CCR3 regulates the Th2-dominant tumor environment, which is closely related to the development of CTCL.

IL-22, but not IL-17, Dominant Environment in Cutaneous T cell Lymphoma

Purpose: Both patients with cutaneous T-cell lymphoma (CTCL) and those with atopic dermatitis (AD) have pruritus, TH2-biased T cells, and a tendency to have bacterial infections, suggesting a common pathologic basis for these two diseases. Recently, interleukin (IL)-22–producing T cells were reported in skin of patients with AD. In this study, we investigated expression levels of TH22- and TH17-related molecules in lesional skin and sera isolated from patients with CTCL. Experimental Design: Skin biopsies and sera were collected from patients with CTCL or psoriasis and from healthy volunteers. Protein and mRNA expression levels of IL-22, IL-17A, IL-17F, IL-23p19, IL-10, IL-4, CCL20, CCR6, IL-8, and IL-20 were examined in lesional tissue and a subset of these molecules in sera. Phosphorylation of STAT3 was also assessed in lesional skin of CTCL and psoriasis by immunohistochemistry. Results: IL-22, IL-10, IL-4, CCL20, and CCR6 mRNA and protein levels, but not IL-17A, IL-17F, IL-23p19, IL-8, or IL-20, were significantly elevated in lesional skin of CTCL. Phosphorylation of STAT3 was detected in epidermis of CTCL skin. Moreover, serum IL-22, IL-10, and CCL20 levels were increased in CTCL and correlated with disease severity. Conclusions: Our results suggest that IL-22 is important in establishing the tumor microenvironment for CTCL. Enhanced expression of CCL20 may explain epidermal hyperplasia and migration of CCR6+ cells, such as Langerhans cells, into lesional skin. Relatively low expression of IL-17 may explain the lack of neutrophils in lesions of CTCL, which correlates with bacterial infections that commonly occur in skin affected by CTCL. Clin Cancer Res; 17(24); 7529–38. ©2011 AACR.

Association of the numbers of CD163+ cells in lesional skin and serum levels of soluble CD163 with disease progression of cutaneous T cell lymphoma

BACKGROUND Classically activated macrophages produce IL-12, IL-23, and TNF-α, whereas alternatively activated macrophages (M2 cells) produce IL-10 and express several receptors such as mannose receptor and CD163. Tumor-associated macrophages exhibit M2 phenotype, whose presence has been associated with poor prognosis in various tumors. OBJECTIVES To investigate distribution of CD163(+) cells in lesional skin and serum levels of soluble CD163 (sCD163) in patients with cutaneous T cell lymphoma (CTCL), atopic dermatitis (AD), or psoriasis. METHODS The numbers of CD163(+) and CD68(+) cells in lesional skin of CTCL, AD, or psoriasis, and in normal skin were examined by immunohistochemistry. Serum soluble CD163 (sCD163) levels were quantified by enzyme-linked immunosorbent assay. RESULTS The numbers of CD163(+) cells in lesional skin of CTCL, AD, or psoriasis were significantly larger than in normal skin. In CTCL, the numbers of CD163(+) or CD68(+) cells increased as more tumor cells infiltrated and they decreased after treatment with topical steroid and ultraviolet light. Moreover, CTCL patients with an increased number of CD163(+) cells showed worse prognosis. Serum sCD163 levels in patients with CTCL, AD, or psoriasis were significantly higher than those in normal controls. In CTCL patients, serum sCD163 levels significantly correlated with serum soluble interleukin-2 receptor and CCL17 levels. In AD patients, serum sCD163 levels correlated with serum IgE levels. CONCLUSION The numbers of CD163(+) cells in lesional skin and serum sCD163 levels were associated with disease progression of CTCL. Further study focusing on CD163(+) cells in CTCL lesional skin would be an interesting research field.

Regulatory B cells in human inflammatory and autoimmune diseases: from mouse models to clinical research

B cells have been generally considered to be positive regulators of immune responses because of their ability to produce antigen-specific antibodies and to activate T cells through antigen presentation. Impairment of B cell development and function may cause inflammatory and autoimmune diseases. Recently, specific B cell subsets that can negatively regulate immune responses have been described in mouse models of a wide variety of inflammatory and autoimmune diseases. The concept of those B cells, termed regulatory B cells, is now recognized as important in the murine immune system. Among several regulatory B cell subsets, IL-10-producing regulatory B cells are the most widely investigated. On the basis of discoveries from studies of such mice, human regulatory B cells that produce IL-10 in most cases are becoming an active area of research. There have been emerging data suggesting the importance of human regulatory B cells in various diseases. Revealing the immune regulation mechanisms of human regulatory B cells in human inflammatory and autoimmune diseases could lead to the development of novel B cell targeted therapies. This review highlights the current knowledge on regulatory B cells, mainly IL-10-producing regulatory B cells, in animal models of inflammatory and autoimmune diseases and in clinical research using human samples.

Increased CCL18 expression in patients with cutaneous T‐cell lymphoma: association with disease severity and prognosis

Background  CC chemokine ligand (CCL) 18 is expressed by monocytes and dendritic cells (DCs), and has potent chemotactic activity for T cells, B cells and DCs. CCL18 expression is up‐regulated in lesional skin of atopic dermatitis and bullous pemphigoid, suggesting its important roles in the development of these skin diseases.

Serum IL-31 levels are increased in patients with cutaneous T-cell lymphoma.

RESULTS As previously reported (4), serum IL-31 levels were significantly higher in patients with AD (2.13 ± 0.26 pg/ ml) than those of healthy controls (0.94 ± 0.28 pg/ml; p < 0.01; Fig. 1a). In addition, serum IL-31 levels were significantly higher in patients with CTCL (3.19 ± 0.74 pg/ml) than those of healthy controls (p < 0.05; Fig. 1a). We subsequently examined serum IL-31 levels in CTCL patients with different types of skin lesions. Serum IL31 levels in patients with patch and plaque, tumour and erythroderma were 1.05 ± 0.30 pg/ml, 6.25 ± 2.11 pg/ml and 5.00 ± 1.46 pg/ml, respectively (Fig. 1b). Serum IL-31 levels in patients with tumour were extremely high, which were significantly higher than those with healthy controls (p < 0.01) and patients with patch and plaque (p < 0.05). Serum IL-31 levels in patients with erythroderma were also significantly higher than those in healthy controls (p < 0.01). Serum IL-31 levels in patients with stage I, stage II, stage III and stage IV were 0.95 ± 29 pg/ml, 3.97 ± 1.43 pg/ml, 2.07 ± 0.00 pg/ ml and 7.98 ± 2.56 pg/ml, respectively (Fig. 1c). Serum IL-31 levels in patients with stage IV were significantly higher than those with healthy controls and patients with stage I (p < 0.05). Serum IL-31 levels in patients with stage II were significantly higher than those with healthy controls (p < 0.05). We also found that serum IL-31 levels correlated significantly with serum sIL-2R and LDH levels (r = 0.43, p < 0.05 and r = 0.34, p < 0.05, respectively; Fig. 1d, e), which have been reported to reflect disease activity of CTCL (8, 9). Thus, serum IL31 levels correlate with disease activity of CTCL.

High Levels of Soluble ST2 and Low Levels of IL-33 in Sera of Patients with HIV Infection

vascular adhesion molecules, or angiogenesis (Figure 2). When comparing C-Tg with C-NT mice, we found significantly more CD3þ cells, CD4þ cells, and CD8þ cells in the Tg strain’s dermis (all with Po0.001). Similarly, the B-Tg dermis had significantly more CD3þ cells (Po0.01), CD4þ cells (Po0.01), and CD8þ cells (Po0.05) compared with its B-NT counterpart. Both C-Tg and B-Tg mice had significantly more PECAMþ vessels per highpower field compared with their NT littermates (Po0.001), further supporting a role of angiogenesis in AD (Chen et al., 2008a). Similarly, C-Tg and B-Tg mice had more P-selectinþ and VCAMþ vessels than did their NT littermates (Po0.001); this was identical to the previous findings in AD patients and in an AD model (Sigurdsson et al., 2000; Chen et al., 2010). There was no difference in numbers of CD3þ and CD4þ lymphocytes, PECAMþ , and P-selectinþ vessels between the dermis of the two Tg strains. The physiological implications of more CD8þ cells and VCAMþ vessels per high-power field in the dermis but with less skin inflammation in C-Tg mice, as compared with B-Tg mice, are unclear (Figure 2b and c). The Th2 systemic immune milieu strongly enhances the rate and extent of cutaneous inflammation in our AD model by primarily increasing the expression of cutaneous Th2 cytokines and other proinflammatory cytokine. CONFLICT OF INTEREST The authors state no conflict of interest.

Erythrodermic cutaneous T-cell lymphoma: How to differentiate this rare disease from atopic dermatitis

Sézary syndrome and erythrodermic mycosis fungoides have been recognized as part of a broader spectrum of erythrodermic cutaneous T-cell lymphoma (E-CTCL). Atopic dermatitis (AD) is the most common, chronic inflammatory skin disease and can, in its most severe form, manifest as erythroderma. It is often difficult to clinically distinguish E-CTCL from various common and benign diseases presenting as erythroderma, including AD. Differentiating E-CTCL from benign inflammatory diseases is important to ensure proper disease management, and to provide accurate prognostic information. Clinical and laboratory features, including pruritus and serum levels of soluble interleukin-2 receptor, lactate dehydrogenase (LDH), immunoglobulin E (IgE), and several chemokines, do not differentiate E-CTCL from AD. In contrast, low serum allergen-specific IgE levels, presence of Sézary cells in peripheral blood, histological findings, and high CD4/CD8 ratio and CCR10 positivity in lesional skin are helpful in reaching a correct diagnosis. Patients with E-CTCL have been treated with oral etretinate, intravenous or subcutaneous interferon, bexarotene, extracorporeal photopheresis, total body surface electron beam, chemotherapy, or any combination of these modalities. Older patients, high serum LDH levels, and high number of circulating atypical lymphocytes are associated with poor prognosis.