Hanako Ohmatsu

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.

Prevalence of atopic dermatitis determined by clinical examination in Japanese adults

Dear Editor, Atopic dermatitis (AD) is an inflammatory skin disease that is characterized by pruritic and eczematous lesions persisting chronically. Studies on the prevalence of AD have produced widely varying figures which may be due to several factors such as subjects’ age, their community and the investigative methodology. There have been numerous studies on the prevalence of AD in children and adolescents, however, there have been few studies on AD in adults. Furthermore, most of these studies were based on hospital patient records or questionnaires. To the best of our knowledge, there has been no study of the prevalence of AD conducted by clinical examination in the general adult Japanese population. The objective of this study was to evaluate the prevalence of AD based on regular health check-ups by dermatologists in Japanese adults. The target population was government officials visiting the Health Service Center of Tokyo University for annual health check-ups in September 2004. Permission was obtained from the Board of the Health Service Center of Tokyo University. The government officials were told the purpose of the study, and those who granted consent participated in this study. AD was diagnosed by experienced dermatologists based on the Japanese Dermatological Association criteria for the disease. The severity of AD was graded as mild, moderate, severe or very severe according to the following criteria: (i) mild, skin involvement of mild eruption only; (ii) moderate, <10% surface area involvement of eruption with severe inflammation (severe eruption); (iii) severe, 710% but <30% skin involvement of severe eruption; and (iv) very severe, >70% of body involvement of severe eruption. The χ test was used to analyze the results, and P < 0.05 was considered statistically significant. A total of 2123 (1220 men and 903 women) officials were examined in this study. The average age was 38.8 ± 10.4 years (men: 39.6 ± 10.5; women: 37.7 ± 10.4) ranging 20–69 years. The prevalence of AD was 6.9% overall, and 9.8%, 8.7%, 4.4% and 2.6%, respectively, for those in their 20s, 30s, 40s and 50s/60s (Table 1). The prevalence of 30s was significantly higher than that of 40s (P < 0.0001). The prevalence of women was higher than that of men overall (9.3% vs 5.1%, P < 0.001), especially in 20s (13.1% vs 5.7%, P < 0.05) and 30s (11.5% 6.9%, P < 0.05). Table 2 depicts the severity of AD. Overall, 76.7%, 18.5%, 3.4% and 1.4% of those afflicted were in the mild, moderate, severe and very severe groups, respectively. There was no severe or very severe AD in those beyond their 40s, nor was there any very severe AD in men. This is the first study of the prevalence of AD determined by clinical examination in the general adult Japanese population. In 1999, Plunkett et al. reported the prevalence of AD based on clinical examination by dermatologists in adults in central Victoria, Australia. A total of 1457 people (670 men and 787 women) whose ages ranged 20–94 years were examined in their study. The prevalence of AD was 6.9% overall, and 5.7% and 8.1%, respectively, for men and women. There was a clear age-related variation: the prevalence was approximately 10%, 8%, 7%, 3%, 2%, respectively, for men in their 20s, 30s, 40s, 50s and 60s, and 22%, 13%, 7%, 7%, 2%, respectively, for women in their 20s, 30s, 40s, 50s and 60s. They classified AD into three categories: mild, moderate and severe. Of those with the disease, 82.8% were classified as being mild, 14.6% moderate, and 2.6% severe. Interestingly, their results and ours suggested the same tendency: (i) the overall prevalence of adult AD was approximately 7%; (ii) the prevalence of women was higher than that of men; (iii) the prevalence decreased with age; and (iv) approximately 80% of AD cases were in the mild group.

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.

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 Gastrin-Releasing Peptide Levels Correlate with Pruritus in Patients with Atopic Dermatitis

effectors of the Wnt pathway, i.e., c-myc and Skp2 were not expressed in KSCs by reverse transcription–PCR (Figure 1b). Further, probing cell lysates of WIF1-arrested keratinocytes in western blots revealed that Wnt3A/ WIF1 treatment resulted in increased p21 protein levels (Figure 2o) demonstrable quantitatively (Figure 2p). Thus, WIF1 may achieve its cell cycle arrest in keratinocytes at least in part through derepression of p21 transcription. In conclusion, we report that WIF1 is, to our knowledge, previously unreported as a marker of interfollicular KSCs, and that it inhibits cell cycle progression in human keratinocytes even in the presence of activating Wnt signals (Wnt3A). Although canonical Wnt signaling appears to be dispensable during development in the interfollicular epidermis (Huelsken et al., 2001; Nguyen et al., 2009), our data suggest that inhibition of Wnt signaling may be required for keeping interfollicular stem cells quiescent and differentiating cells from proliferating during homeostasis.

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.

A Randomized, Open-Label, Multicenter Trial of Topical Tacrolimus for the Treatment of Pruritis in Patients with Atopic Dermatitis

Background Pruritis caused by atopic dermatitis (AD) is not always well controlled by topical corticosteroid therapy, but use of tacrolimus often helps to soothe such intractable pruritis in clinical settings. Objective To determine the anti-pruritic efficacy of topical tacrolimus in treating AD in induction and maintenance therapy. Methods Prior to the study, patients were randomly allocated into two groups, induction therapy followed by tacrolimus monotherapy maintenance, and induction therapy followed by emollient-only maintenance. In the induction therapy, the patients were allowed to use topical tacrolimus and emollients in addition to a low dose (<10 g/week) of topical steroids. Patients showing relief from pruritis were allowed to proceed to maintenance therapy. Recurrence of pruritis in maintenance therapy was examined as a major endpoint. Results Two-thirds of patients (44/68; 64.7%) showed relief from pruritis after induction therapy. Pruritis recurred in 23.8% (5/21) of the tacrolimus monotherapy group and in 100% (21/21) of the emollient group during maintenance period, a difference that was statistically significant. Conclusion Use of topical tacrolimus is effective in controlling pruritis of AD compared to emollient.

TOX expression in different subtypes of cutaneous lymphoma

Early cutaneous T cell lymphoma clinically and histologically resembles benign inflammatory skin diseases, which sometimes makes it difficult to reach a correct diagnosis. It is recently reported that thymocyte selection-associated high mobility group box factor (TOX) serves as a molecular marker for histological diagnosis of early-stage mycosis fungoides (MF). To examine whether TOX could be a marker of tumour cells in different types of cutaneous lymphoma, we investigated immunohistochemical staining for TOX with the lesional skin of patch, plaque, and tumour MF, Sézary syndrome (SS), lymphomatoid papulosis (LyP), primary cutaneous anaplastic large cell lymphoma (PCALCL), adult T cell leukemia/lymphoma (ATLL), peripheral T-cell lymphoma, not otherwise specified (PTCL, NOS), atopic dermatitis (AD), and normal skin. TOX and CCR4 messenger RNA (mRNA) levels in lesional skin of MF/SS were also examined. Immunohistological staining showed that a high specific nuclear staining of TOX was observed at a high frequency in MF, SS, and PTCL, NOS. Tumour cells in LyP, PCALCL, and ATLL showed a slightly dim nuclear staining of TOX. TOX+ cells in MF and LyP expressed surface molecules characteristics of tumour cells in these diseases. Lesional skin of SS expressed higher levels of TOX mRNA, compared to normal skin or MF lesional skin. Moreover, TOX expression significantly correlated with CCR4 expression. TOX may be a specific marker for tumour cells in some types of cutaneous lymphoma.