Successful treatment of lichen amyloidosus associated with atopic dermatitis using a combination of narrowband ultraviolet B phototherapy, topical corticosteroids and an antihistamine.

Lichen amyloidosus (LA) is a type of primary localized cutaneous amyloidosis characterized by multiple pruritic discrete hyperkeratotic papules with amyloid deposition in the papillary dermis. Clinical regression is usually difficult to achieve, even after treatment. In this study, we report a case of an adult man with LA associated with atopic dermatitis (AD) which was successfully treated with narrowband ultraviolet B (NB-UVB) phototherapy, topical corticosteroids and an oral antihistamine. This case suggests that NB-UVB phototherapy may be a useful adjuvant for LA associated with AD.

Epinastine inhibits eosinophil chemotaxis and adhesion molecules in atopic dermatitis.

PURPOSE: To investigate the effects of epinastine on eosinophil chemotaxis and changes in eosinophil adhesion molecules induced by epinastine and three other antiallergic agents, using eosinophils of atopic dermatitis (AD) patients. RESULTS: Epinastine reduced eosinophil chemotaxis toward eotaxin when the eosinophils had been prestimulated with interleukin (IL)-5, but given alone it did not alter eosinophil chemotaxis toward IL-5. CD11b expression was inhibited when peripheral blood was prestimulated with IL-5, but eosinophil adhesion molecule expression was not altered. CONCLUSIONS: Epinastine suppresses allergic inflammation not only through its strong antihistamine and antimediator effects, but also by inhibiting eosinophilic chemotaxis and the expression of adhesion molecules involved in chemotaxis, especially CD11b.

Expression of NF-kappaB in epidermis and the relationship between NF-kappaB activation and inhibition of keratinocyte growth.

BACKGROUND: Nuclear factor-kappaB (NF-kappaB) is a transcription factor involved in a number of signalling pathways in many cell types. NF-kappaB in mice has been implicated as an important regulator of keratinocyte proliferation and differentiation. OBJECTIVES: To evaluate the role of NF-kappaB in keratinocyte growth in human beings, we examined its expression in keratinocytes both in culture and in situ, and studied the relationship between NF-kappaB activation and the inhibition of keratinocyte proliferation induced by known modulators of keratinocyte growth. METHODS: The expression of subunits of the NF-kappaB family was examined in human skin, primary cultured keratinocytes and an immortalized keratinocyte line by immunohistochemistry and reverse transcriptase-polymerase chain reaction analysis. NF-kB activation was examined in keratinocytes treated with various modulating agents by electrophoretic mobility shift assay (for DNA-binding activity) and by immunocytochemistry (nuclear translocation). The proliferative capacity of treated keratinocytes was also examined by 3H-thymidine incorporation, cell cycle analysis, and expression of Ki-67, a nuclear marker for cell proliferation. The involvement of NF-kappaB was assessed using sodium salicylate, which inhibits NF-kappaB activation. RESULTS: The NF-kappaB subunits, p50, p65, RelB, and c-Rel (but not p52), were detected in keratinocytes and in normal epidermis at mRNA and protein levels. The four subunits were expressed in a cytoplasmic (rather than a nuclear) pattern in both basal and suprabasal keratinocytes. Phorbol myristate acetate (PMA), tumour necrosis factor alpha, and interferon gamma each activated NF-kappaB and inhibited keratinocyte proliferation. Lipopolysaccharide and dexamethasone did not activate NF-kappaB and had the least effect on proliferation. Finally, a high concentration of calcium (Ca2+) and retinoic acid each failed to activate NF-kappaB, but were potent inhibitors of keratinocyte proliferation, respectively. PMA-induced cell cycle arrest of keratinocytes was blocked by pretreatment with sodium salicylate. CONCLUSIONS: NF-kappaB is constitutively expressed in a resting state in both human cultured keratinocytes and the epidermis. Activation of NF-kappaB is required for PMA-induced keratinocyte growth arrest.

The successful treatment of diffuse neonatal hemangiomatosis with flashlamp pulsed dye laser.

Diffuse neonatal hemangiomatosis is a rare disease with the distinctive features of multiple hemangiomas of the skin and visceral organs. These lesions have been treated with systemic corticosteroids, interferon-alpha, and their combination. We report a patient with diffuse neonatal hemangiomatosis who had multiple cutaneous and hepatic hemangiomas. Single therapy with the flashlamp pulsed dye laser was effective for cutaneous hemangiomas, whereas the hemangiomas of the liver remained stable and no liver dysfunction or hemorrhage has occurred so far, even with no treatment.

Overexpression of CD11b on eosinophils in atopic dermatitis: downregulation by cyclosporin A and upregulation by interleukin 5.

We examined the level of expression of CD11b on eosinophils in pripheral blood samples from patients with atopic dermatitis (AD) and non-AD volunteers. Eosinophils were defined using a new method employing CD14/CD45 and a backgate technique. Overexpression of CD11b was noted in eosinophils of AD patients. Treatment of AD with cyclosporin A resulted in clinical improvement as well as reduction in the expression of CD11b. Stimulation of eosinophils from patients with inactive AD by interleukin 5 upregulated the expression of CD11b on these cells. Our results suggest that the expression of CD11b surface molecule on eosinophils may play an important role in the activity of AD.

Enhanced production of RANTES, an eosinophil chemoattractant factor, by cytokine-stimulated epidermal keratinocytes.

In allergic skin diseases such as atopic dermatitis (AD), eosinophils migrate from the circulation to the skin. We investigated the mechanisms of eosinophil chemotaxis in atopic dermatitis by examining the effect of stimulation of epidermal keratinocytes (KC) by inflammatory cytokines, interferon-gamma (IFNgamma) and/or tumor necrosis factor-alpha (TNF alpha) on the production of eosinophil chemotactic factors. Simultaneous addition of IFNgamma and TNF alpha to culture KC synergistically increased eosinophil chemotaxis and the expression of RANTES mRNA and protein level on these cells. Anti-RANTES antibody blocked eosinophil chemotaxis by IFNgamma- and TNF alpha-stimulated KC. Our results indicate that the production of RANTES by KC may help to explain eosinophil infiltration into the skin in AD.

Steroid-induced changes of eosinophils in atopic dermatitis.

We investigated whether apoptosis of eosinophils is specific to atopic dermatitis (AD), or also occurs in other diseases with eosinophilia. We examined the survival of eosinophils cultured with corticosteroids: (1) Clinically, steroid administration significantly decreased high peripheral blood eosinophil cell counts in patients with AD. (2) Treatment with recombinant human (rh) IL-5 prolonged the life span of eosinophils derived from patients with AD and of those derived from non-AD patients with eosinophilia. However, there were differences in the survival rates in the presence of rhIL-5: the eosinophils from non-AD patients showed 1.4-fold higher survival rates than those from AD patients at 24 h. In the presence of steroids, the eosinophils from non-AD patients showed a survival rate double that of those from AD patients at 24 h. (3) In eosinophils from patients with AD, the survival rate decreased significantly in a time- and steroid-concentration-dependent manner. Steroid administration significantly inhibited the survival rate of eosinophils from patients with AD compared to those of monocytes and neutrophils. These findings suggest that apoptosis induced by steroids decreases the eosinophil count in vivo in patients with AD. There may be a difference in the incidence of steroid-induced apoptosis between eosinophil cells from patients with AD and those from patients with eosinophilia due to other underlying diseases.

Immunohistology of skin and oral biopsies in graft-versus-host disease after bone marrow transplantation and cytokine therapy.

BACKGROUND: Early diagnosis of graft-versus-host-disease (GVHD) after bone marrow transplantation is often difficult, particularly when the patients are immunosuppressed by chemotherapy or irradiation. OBJECTIVE: To investigate the influence of cytokines on skin lesions after bone marrow transplantation. METHODS: Biopsy specimens of skin and oral mucosa were obtained from bone marrow transplant patients with GVHD and were subjected to histologic and immunohistochemical examination. RESULTS: Administration of granulocyte-macrophage colony-stimulating factor caused atopic dermatitis-like lesions in two patients, who had infiltration of neutrophils, eosinophils, and lymphocytes around the hair follicles of the skin and no signs of GVHD in other organs. Only patients who were treated with cytokines developed acute GVHD. Immunohistochemical examination of skin biopsies from 18 patients with acute GVHD and 11 patients with chronic GVHD after cyclophosphamide administration or irradiation showed that the maculopapular skin lesions characteristic of acute GVHD contained infiltrates of CD4+ and CD8+ lymphocytes. There was also an increase in numbers of epidermal keratinocytes expressing intercellular adhesion molecule-I and HLA-DR antigens. CONCLUSION: These findings support the involvement of cytokines in GVHD and suggest that immunostaining of skin biopsies may be useful for the early diagnosis of this condition.

Role of staphylococcal enterotoxins in pathogenesis of atopic dermatitis: growth and expression of T cell receptor V beta of peripheral blood mononuclear cells stimulated by enterotoxins A and B.

In order to investigate the role of staphylococcal enterotoxins in the pathogenesis of dermatitis in atopic patients, the growth and expression of T cell receptor V beta in peripheral blood mononuclear cells (PBMCs) from atopic dermatitis patients induced by stimulation with staphylococcal enterotoxin A (SEA) or staphylococcal enterotoxin B (SEB) were examined. Lymphocyte stimulation tests (LST) using SEA or SEB were performed in atopic dermatitis (AD) patients (n = 10) and normal controls (n = 5). PBMCs from AD patients displayed significantly stronger responses to SEA or SEB than those from the controls. To ascertain further whether SEA acts as a superantigen in atopic dermatitis, the expression of 22 genes in the variable region of the beta chain (V beta) of T cell receptors (TcR) was examined before and after stimulation with SEA by a reverse transcriptase-polymerase chain reaction (RT-PCR). Before stimulation, only weak expression of V beta was observed, and the expression of the various V beta segments was uniform in the normal controls (n = 3). In the AD patients (n = 3), the expression of V beta was enhanced, but was not uniform in 2 out of 3 patients and the pattern of expression was characteristic in each individual. This suggests that V beta expression varies in individual AD patients and displays restricted heterogeneity, reflecting the diversity of the etiology of the disease. After culture of the SEA-stimulated cells, no difference was observed in the expression of TcR V beta segments in the 3 normal controls as compared with that prior to stimulation, but particular V beta segments were intensely expressed in 3 AD patients, displaying distinct patterns (case I: V beta 9, V beta 10, V beta 18; case 2: V beta 6.1-3; case 3: V beta 6.1-3, V beta 18). Many of these V beta segments corresponded with those known to be induced by SEA. These results suggest oligoclonal proliferation of T cells in the peripheral blood of AD patients and high responsiveness in each clone, and since the expression of V beta segment after SEA stimulation was restricted, the actions of staphylococcal enterotoxins as superantigens were suggested.

[Analysis of the mechanism of trophoblast infiltration].

A cytotrophoblast (CT) infiltrates into the stroma, forming an extravillous trophoblast (EVT) in the placenta early in gestation and the phenomenon is strictly controlled, differing from the infiltration of cancer cells. The expression of matrix metalloproteinase 2 (MMP2) and matrix metalloproteinase 9 (MMP9), which deeply involve infiltrative metastasis of cancer, and the reactivity to transforming growth factor beta 1 (TGF beta 1), which controls the expression of these MMPs and inhibits the growth of epithelial cells, were investigated in CT derived from villi at normal gestational week 6 (early CT) and CT derived from villi at normal gestational week 37 (full-term CT), and also the choriocarcinoma cell line BeWo (BeWo). The ability of normal epithelial cells and BeWo cells to proliferate and infiltrate were evaluated in vitro by northern blotting, gelatin zymography, and invasion assay. It was revealed that early CT had a higher capacity for infiltration than full-term CT as well as BeWo. MMP2 and MMP9 appeared in the early CT, whereas only MMP9 was observed in the full-term CT. MMP2 and MMP9 were more abundantly observed in the early CT and the full-term CT rather than in BeWo. In uterine stroma-derived cells, membrane type matrix metalloproteinase (MT-MMP), which activates MMP2, was observed. These results indicated that the motility of normal villous cells was higher in the early CT than in the full-term CT. The expression of MMP2 in the early CT, which was not observed in the full-term CT, was thought to be related to this difference in motility. As for the responsiveness to TGF beta 1, which is a growth inhibiting factor for epithelial cells, the villous carcinoma cell line was insensitive to the growth inhibiting effect of TGF beta 1, but the early CT was sensitive to this effect. When TGF beta 1 was added, MMP2 and MMP9 increased in the early CT. This response was also seen in BeWo. That is, it was assured that the growth capacity was not inhibited in BeWo, but was certainly inhibited in the early CT. The overall results of these evaluations indicated that the development to EVT by infiltration of the early CT was associated with the increase in the mobility of cells caused by MMP2 and the increase in amounts of MMP2 and MMP9 caused by TGF beta 1, and the predominant inhibitory effect of TGF beta 1 on the growth of normal epithelial cells could explain why normal epithelial cells do not grow as cancer cells do.

Elevated plasma RANTES levels in patients with atopic dermatitis.

Eosinophils and T cells are involved in the pathologic process of atopic dermatitis. To further understand the role of these cells, and the possible involvement of RANTES, in the pathogenesis of atopic dermatitis, we measured the plasma level of RANTES using the sandwich ELISA. The mean plasma level of RANTES in 11 patients with atopic dermatitis was significantly higher than that of 15 normal control subjects. RANTES levels were higher in patients with severe form of atopic dermatitis than that of patients with mild disease. These findings suggest that RANTES may play a role in the recruitment and activation of eosinophils and T cells in atopic dermatitis.

RANTES mRNA expression in skin and colon of patients with atopic dermatitis.

The expression of RANTES mRNA in dermal and colonic tissue was examined in patients with atopic dermatitis by the reverse transcription polymerase chain reaction method. RANTES mRNA was detected in the colon in 8 of 10 patients and in 1 of 5 control patients. It was present in rashes in 9 of 10 patients and at non-eruptive sites in 5 of 7 patients. These findings suggest that RANTES is involved in eosinophil infiltration and T cell infiltration in atopic dermatitis.

Corticosteroid-induced apoptosis of eosinophils in atopic dermatitis patients.

We examined the mechanism by which steroid administration significantly decreases the high eosinophil cell count in peripheral blood in patients with atopic dermatitis (AD). Eosinophils were isolated from the peripheral blood of patients with moderate or severe adult AD, and cultured. After steroid was added to the culture medium, we examined the changes in eosinophils, i.e., 1) the survival rates, 2) morphological changes and 3) fragmentation of DNA with respect to 2 factors, the steroid concentration and culture time. The steroid or interleukin-5 (IL-5) were added to eosinophils from patients with AD and those from non-AD patients with eosinophilia to compare serial changes in the survival rate. In eosinophils from patients with AD, the survival rate significantly decreased time-dependently and steroid concentration-dependently. Steroid administration significantly inhibited the survival rate of eosinophils from patients with AD compared to the survival rates of monocytes and neutrophils. The nuclei of eosinophils were serially reduced, and disappeared 72 hours after steroid administration. Simultaneously, cell size decreased, although the cell membrane remained intact. Granules developed in the cell membrane. In the steroid-treated group, apoptotic cells appeared earlier than in the untreated group. The number of cells showing apoptosis was increased steroid concentration-dependently. The number of DNA ladders was increased time-dependently and steroid concentration-dependently. In eosinophils derived from patients with AD and those derived from non-AD patients with eosinophilia, treatment with recombinant human (rh) IL-5 prolonged the life-span of cells. However, there were differences in the survival rates. In the presence of rhIL-5, the eosinophils from non-AD patients survived 1.4 times longer than those from AD patients at 24 hours (P < 0.05). In the presence of steroid, the eosinophils from non-AD patients survived twice as long as those from AD patients at 24 hours (P < 0.01). These findings suggest that apoptosis induced by steroids decreases the eosinophil count in vivo in patients with AD. There may be a difference in the incidence of steroid-induced apoptosis between eosinophil cells from patients with AD and those from patients with eosinophilia due to other underlying diseases.

Serum levels of anti-Staphylococcus aureus-specific IgE in patients with atopic dermatitis.

We investigated the role of Staphylococcus aureus-specific IgE in patients with atopic dermatitis (AD). The titer of serum S. aureus-specific IgE was measured using the RAST method in 67 patients with AD and correlated with serum LDH, eosinophil count and total IgE. The titer of S. aureus-specific IgE was elevated in 41 patients but was not detected in 26 patients. The mean serum level of total IgE was higher in the positive group than in the negative group, but the eosinophil count and LDH levels were not different between the two groups. S. aureus was detected and cultured from the skin of 33/41 (80%) patients in the positive group, but only from the skin of 5/26 (19%) patients of the negative group. Our results suggest that S. aureus-specific antibody is present in patients with moderate-to-severe atopic dermatitis and may be involved in the pathogenesis of AD.

The POU domain transcription factor Brn-2 is required for the determination of specific neuronal lineages in the hypothalamus of the mouse.

We generated mice carrying a loss-of-function mutation in Brn-2, a gene encoding a nervous system specific POU transcription factor, by gene targeting in embryonic stem cells. In homozygous mutant embryos, migratory precursor cells for neurons of the paraventricular nuclei (PVN) and the supraoptic nuclei (SO) of the hypothalamus die at approximately E12.5. All homozygous mutants suffered mortality within 10 days after birth, possibly because of a complete deficiency of these neurons in the hypothalamus. Although neither developmental nor histological abnormalities were observed in heterozygous mice, the levels of expression of vasopressin and oxytocin in the hypothalamus of these animals were half these of wild-type mice. These results strongly suggest that Brn-2 plays an essential role in the determination and development of the PVN and SO neuronal lineages in the hypothalamus.

[mRNA expression and protein localization of placental tissue protein 11, 12, 19 in gynecologic malignant tumors].

In recent years, the localization and function of placental tissue proteins (PPs), extracted by Bohn et al., have been extensively studied, the genetic code has been identified for each of the PPs. The present study was carried out to clarify the mRNA expression and protein localization of PP11, PP12, PP19 at the cell level and also to define PP19 the nature of which has remained obscure. PP19 was said to be placenta-derived S-100P. PP11 mRNA was not expressed in cytotrophoblast-derived normal placental tissue or endometrium-derived normal cells, but was expressed in syncytiotrophoblast-derived normal placental tissue, and in choriocarcinoma and endometrial adenocarcinoma, suggesting its involvement in carcinogenesis. PP12 mRNA was expressed in cytotrophoblast-derived normal placental tissue or endometrium-derived normal cells, but was not expressed in syncytiotrophoblast-derived normal placental tissue, choriocarcinoma or endometrial adenocarcinoma, suggesting that this protein serves in the function of normal cells. PP19 mRNA was expressed in the squamous epithelial cells of the uterine cervix and the villous cells, PP19 was localized in more differentiated regions, where cells tended toward keratinization, in both normal and dysplastic uterine cervices. In squamous cell carcinoma, PP19 was localized in more differentiated cells with a large cytoplasm. PP19 mRNA was not expressed in normal endometrial glands, but was detected in endometrial adenocarcinoma, suggesting its involvement in cell differentiation in cervical epitherial cells.