Hiromichi Kai

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.

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.

CCR10 and CCL27 are overexpressed in cutaneous squamous cell carcinoma.

C-C chemokine receptor (CCR)10 is a specific receptor for chemokine ligand (CCL)27, a selective chemoattractant for skin-associated memory T cells to cutaneous sites. In melanoma, CCR10 increases the ability of neoplastic cells to grow, invade tissues, disseminate to lymph nodes, and escape the host immune responses. In this study, we investigated the expression of CCR10 and its ligand CCL27 in squamous cell carcinoma (SCC). CCR10 and CCL27 were expressed in SCC, actinic keratosis (AK), Bowens disease, and seborrheic keratosis (predominantly prickle cell type), but not in seborrheic keratosis (predominantly basal cell type) and basal cell carcinoma. Furthermore, CCR10 and CCL27 were overexpressed in SCC relative to Bowens disease, an early stage of SCC. Consistently, a human SCC cell line, A253 cells, and HaCaT cells exhibited CCL27 production that was strongly induced by tumor necrosis factor-α and interleukin-1β. Finally, A253 cells expressed stronger intracellular CCR10 compared to HaCaT cells by flow cytometry. These results suggest that CCR10 and CCL27 overexpression in SCC is related to the progression of SCC and is useful for the diagnosis of SCC.

Variations in serum TARC and I-TAC levels reflect minor changes in disease activity and pruritus in atopic dermatitis.

Atopic dermatitis (AD) is a chronic or relapsing inflam-matory skin disease. Scratching in AD patients results in proinflammatory cytokine and chemokine production. Thus, serum levels of monocyte chemotactic protein-1 (MCP-1), regulated on activation, normal T-cell expressed and secreted (RANTES ), macrophage inflammatory pro-tein (MIP)-1β, eotaxin, thymus and activation-regulated chemokine (TARC), and macrophage-derived chemokine (MDC) were increased in AD patients, compared with normal controls (1–4). With regards to Th1 chemokines such as interferon (IFN)-γ-inducible protein 10 (IP-10), IFN-γ-inducible T-cell α-chemoattractant (I-TAC), and monokine induced by IFN-γ (MIG), their expression in le-sional AD skin was confirmed by immunohistochemistry ±(5). They may negatively contribute to the development of AD because macrophages from AD patients produced lower levels of IP-10 compared to cells from healthy con -trols in response to α-toxin (6) and the expression of MIG and IP-10 was lower in Langerhans cells from patients with AD than from patients with psoriasis, whereas the opposite was observed for TARC and MDC (7).Visual analogue scale (VAS) is a valuable method to assess pruritus intensity in patients with pruritic der-matoses (8). In this study , we focus on temporal variation of pruritus in each patient and compare serum samples taken at different time points when there were only slight, if any, changes in disease activity. The aim of this study was to highlight the most sensitive chemokine associated with changes in pruritus in AD patients. MATERIALS AND METHODS

Analysis of serum chemokine levels in patients with HIV-associated eosinophilic folliculitis

Background  Patients with human immunodeficiency virus (HIV) infection exhibit various skin diseases. HIV‐associated eosinophilic folliculitis (EF) and pruritic papular eruption (PPE) are frequently seen.

Extensive skin pigmentation caused by deposits of metallic particles following total elbow arthroplasty: metallosis or not?

patients with localized psoriasis of palms and soles, generalized pustular psoriasis and acrodermatitis continua, conditions which were associated with a high incidence of arthritis. This may suggest that our patient had a psoriatic background, developing acrodermatitis continua of Hallopeau, which was induced by oral terbinafine. Oral terbinafine, therefore, should not be considered as first-line therapy for onychomycosis in psoriatic individuals. F . N I SH IWAK I Y . MAT SUMURA N. MOR I TA S . KORE -EDA Y. MIYACH I M. OMOTO* Department of Dermatology, Graduate School of Medicine, Kyoto University, 54 Shogoin, Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan *Department of Dermatology, Hikone Municipal Hospital, 1882 Yasaka-cho, Hikone-city, Shiga 522-8539, Japan E-mail: fuyuko@kuhp.kyoto-u.ac.jp

Depsipeptide and roxithromycin induce apoptosis of lymphoma cells by blocking extracellular signal-regulated kinase activation

Depsipeptide (FK228), a histone deacetylase inhibitor, was recently approved for use in cutaneous T‐cell lymphoma. Roxithromycin (RXM) is a macrolide antibiotic that can induce apoptosis of some T‐cell lines. In this study, we investigated whether combination of FK228 and RXM had a synergistic inhibitory effect on cell survival of various lymphoma cells and which signaling pathway was affected by the drugs in the presence or absence of chemokines, which were reported to inhibit apoptosis of some tumor cells. FK228 and RXM additively decreased the number of HUT‐78, Ki‐JK and EL‐4 lymphoma cells at doses over 50 nmol/L and 50 μmol/L, respectively. These drugs inhibited phosphorylation of Akt and extracellular signal‐regulated kinase (ERK) of EL‐4 cells in a dose‐dependent manner. Significant association between ERK phosphorylation and cell number or annexin V+ cells suggested that the ERK pathway may be critical for survival of EL‐4 cells. Combination of 10 or 50 nmol/L of FK228 and 10 μmol/L of RXM decreased cell number of HUT78 and EL‐4 compared to a single use of each drug. Our in vitro study suggested that combination of FK228 and RXM may be helpful for enhancing tumor killing effects. Although further study is necessary, this combination may be applicable to patients with cutaneous T‐cell lymphoma in the future.

High Levels of CCL26 in Blister Fluid and Sera of Patients with Bullous Pemphigoid

necrosis factor-a. Out of 160 genes, 53 ‘‘synergistic’’ genes were identified in at least one of the aforementioned published studies; our RNA-seq analysis identified 14 additional upregulated DEGs (Supplementary Table S2 online), further emphasizing the contribution of this immunological circuit in the pathogenesis of psoriasis. One of these genes was SPRR2B; staining for the protein product of this gene confirmed increased expression in psoriatic lesional skin (Figure 2c), corroborating differences at the protein level of a gene identified uniquely by RNA-seq that is potentially informative with regard to active molecular pathways in psoriasis. The transcriptomic profiling of psoriasis has led to an increased understanding of disease pathogenesis. Although microarray technologies have been instrumental in this regard, it is clear that these tools detect an incomplete set of DEGs. RNA-seq can be used to supplement these prior technologies. Here, the use of RNAseq methods substantially increased the number of psoriasis-related DEGs. Furthermore, DEGs that were uniquely identified by RNA-seq, but not in other published microarray studies, further supported the role of IL-17 and tumor necrosis factor-a synergy in psoriasis. Examination of one of these factors at the protein level confirmed that RNA-seq is a powerful tool that can be used to identify molecular factors present in psoriasis lesions, and may be useful in the identification of therapeutic targets that to our knowledge have not been reported previously. Further studies are in progress to determine the biological significance of DEGs uniquely discovered by RNA-seq.

Elevated serum galectin‐9 levels in patients with atopic dermatitis

Galectin‐9 is a member of the galectin family that has a wide spectrum of biological functions. Among them, galectin‐9 has been known mainly as a potent chemoattractant for eosinophils. In addition, galectin‐9 alters the T‐cell balance by negatively regulating T‐helper (Th)1 and Th17 cells, resulting in Th2 polarization. Atopic dermatitis (AD) is a skin allergic disease characterized by peripheral eosinophilia, mast cell activation and predominance of Th2 cells. To investigate possible roles of galectin‐9 in AD, we measured serum galectin‐9 levels in AD patients and investigated galectin‐9 expression in lesional skin by immunohistochemistry. Serum galectin‐9 levels in patients with AD were significantly higher than those in healthy controls and correlated with the Eczema Area and Severity Index. Serum galectin‐9 levels were decreased after treatment, accompanied by improvement of skin lesions. Immunohistochemical study revealed that galectin‐9 was expressed on epidermal keratinocytes and mast cells in lesional skin of AD. Our results suggest that elevated galectin‐9 expression is associated with progression of AD and that galectin‐9 could be a therapeutic target in AD.