他克莫司联合NB-UVB对小鼠IL-10表达的影响

目的:确定他克莫司联合应用NB-UVB对小鼠IL-10表达的影响.方法:30只BALB/c小鼠随机分为空白对照组、他克莫司组、他克莫司+NB-UVB组、NB-UVB+他克莫司组及单用NB-UVB组.采用酶联免疫吸附试验(ELISA)检测皮肤及血液中IL-10的含量.结果:单用NB-UVB、他克莫司+NB-UVB、NB-UVB+他克莫司以及单用他克莫司组的皮肤及血液IL-10的含量与正常对照组的差异均有显著性(P＜0.05).结论:单用NB-UVB或他克莫司及二者联合应用均可使血清及皮肤组织悬液中IL-10含量升高,但联合应用对小鼠局部及系统免疫抑制无协同作用.     

茶多酚、吡美莫司、他克莫司外用对莫诺苯宗诱导白癜风样模型小鼠的疗效比较

目的 探讨茶多酚、吡美莫司、他克莫司对莫诺苯宗诱导的白癜风样模型小鼠的疗效差异。 方法 45%莫诺苯宗诱导C57BL/6三周龄雌小鼠脱色,建立白癜风样动物模型。并研究他克莫司、吡美莫司、茶多酚对白癜风样模型小鼠脱色的治疗效果。通过肉眼观察毛发脱色及脱色面积,实验结束后取非用药部位脱色皮肤行组织学检查,HE染色检测淋巴细胞浸润情况,共聚焦激光扫描显微镜（RCM）观察小鼠皮肤的黑素和黑素细胞,免疫荧光检测CD8+ T细胞 。 结果 模型组小鼠在用药部位及非用药部位均有脱色现象。他克莫司组、吡美莫司组、茶多酚组小鼠脱色减少,出现时间晚和面积指数均较模型组低;且用药部位脱色斑局部淋巴细胞和CD8+ T细胞浸润减少。其中他克莫司组疗效优于其他组。 结论 他克莫司、吡美莫司、茶多酚对白癜风样模型小鼠均有疗效。     

他克莫司影响朗格汉斯细胞迁移机制的研究

目的探讨他克莫司影响朗格汉斯细胞迁移能力的原因。方法用不同浓度的他克莫司处理人角质形成细胞株HaCaT，24h后收集上清液，通过ELISA法检测上清液中的单核细胞化学吸引蛋白质1(monocyte chemoattractant protein 1，MCP—1)浓度，并通过RT—PCR法检测细胞中MCP—1 mRNA水平。结果经质量浓度为625～5000ng／mL他克莫司处理过的HaCaT株，其分泌至上清液中的MCP-1水平明显受到抑制，细胞中MCP-1 mRNA的水平低于正常对照组。结论他克莫司通过抑制角质形成细胞中MCP-1的分泌、表达，进而影响了朗格汉斯细胞前体及朗格汉斯细胞趋化至皮损处。阻止或减弱了某些免疫反应。     

他克莫司治疗口腔扁平苔藓20例疗效评价

为了评价0.1%他克莫司软膏治疗口腔扁平苔藓(OLP)的疗效与安全性,我们采用0.1%他克莫司软膏外用治疗20例OLP,以曲安奈德局部注射治疗作对照.治疗组有效率80%,对照组有效率75%,两组疗效差异无显著性(P>0.05).治疗组2例出现局部轻度灼痛感,不需治疗.0.1%他克莫司软膏外用治疗OLP与曲安奈德局部注射疗效相当,有较高的安全性.     

强脉冲光联合他克莫司软膏治疗面部脂溢性皮炎疗效观察

目的探讨强脉冲光联合他克莫司软膏治疗面部脂溢性皮炎的疗效.方法治疗组24例患者应用强脉冲光照射,每3周治疗1次,3次为1个疗程,同时外用0.03％他克莫司软膏,每日2次.对照组21例仅外用0.03％他克莫司乳膏,每日2次,共4周.两组均于治疗结束后半个月判定疗效.结果两组瘙痒、红斑、鳞屑均有明显改善,治疗组和对照组临床有效率分别为87.50％和57.14％,治疗组疗效优于对照组.外用他克莫司软膏的7例患者局部有一过性刺激反应.治疗组面部经表皮丢失水分、皮肤油脂、皮肤红斑指数、皮肤弹性较治疗前有明显改善,差异有统计学意义(P＜0.05).结论强脉冲光联合外用他克莫司软膏治疗面部脂溢性皮炎安全、有效.     

他克莫司软膏治疗特应性皮炎疗效和安全性评价

目的：系统评价外用他克莫司软膏治疗特应性皮炎（AD）的临床疗效及安全性。方法：计算机检索Cochrane图书馆、Cochrane协作网皮肤病专业试验数据库、Medline、OVID数据库和中文生物医学期刊数据库．收集所有外用他克莫司与安慰剂、氢化可的松的随机对照试验（RCT）,对其进行系统评价。结果：共纳入RCT13篇论文,共5320例患者。Meta分析治疗有效率,结果显示：0．03％和0．1％他克莫司在12周疗程内疗效均优于安慰剂;0．03％和0．1％他克莫司3周疗程均高于1％醋酸氢化可的松,均不高于0．1％丁酸氢化可的松,但0．1％他克莫司在6个月疗程时疗效优于合用1％醋酸氢化可的松（用于头面部）和0．1％丁酸氢化可的松（用于躯干和四肢）;0．1％他克莫司在疗程12周内疗效优于0．03％他克莫司。最常见的不良反应是皮肤刺激和烧灼感。结论：他克莫司软膏治疗AD效果优于安慰剂及弱效糖皮质激素,长期疗效可能超过中强效糖皮质激素。目前外用他克莫司临床上是安全的,但尚需进行更多长期的RCT。     

外用他克莫司与卡泊三醇治疗斑块状银屑病的疗效与安全性评价

目的:评价外用他克莫司和卡泊三醇软膏对四肢斑块状银屑病的疗效和安全性.方法:选取门诊斑块状银屑病患者,四肢左右两侧分别外用他克莫司软膏及卡泊三醇软膏,治疗2周及6周各复诊1次,观察四肢左右侧皮损消退情况,用单侧银屑病皮损面积及严重度指数(PASI)评分进行评价.结果:29例患者参加自身左右侧药物对比,有3例患者(10.3％)应用他克莫司后出现皮肤刺激症状而停用,其中1例患儿出现红皮病样改变.2周时有13例患者复诊,达到PASI50的患者中他克莫司侧4例,卡泊三醇侧5例；达到PASI75的患者中他克莫司侧1例,卡泊三醇侧2例.6周时14例患者复诊,达到PASI50的患者中他克莫司侧14例,卡泊三醇侧13例；达到PASI75的患者中他克莫司侧7例,卡泊三醇侧9例.2周及6周时比较双侧PASI50及PASI75,用Fisher精确概率检验法,发现两者差异无统计学意义.结论:对于四肢的斑块状银屑病,外用他克莫司和卡泊三醇软膏均有肯定疗效.部分患者应用他克莫司出现皮肤刺激症状,停药后症状多可消退.     

外用他克莫司和吡美莫司治疗红斑狼疮皮肤损害的研究进展

他克莫司和吡美莫司是两种新的钙调神经磷酸酶抑制剂,皮肤红斑狼疮的免疫源性的病因假说提示了他克莫司和吡美莫司有效的可能.临床报道他克莫司和吡美莫司治疗红斑狼疮皮肤损害有一定疗效.     

外用他克莫司软膏治疗白癜风临床观察

目的：观察他克莫司软膏治疗白癜风的临床疗效及安全性。方法：给予33例白癜风患者他克莫司软膏，每日2次外用，观察治疗后靶皮损复色情况，并与治疗前进行对比。结果：33例患者共计81处皮损，外用他克莫司软膏1～4个月后有效率为60．49％。2例患者用药后出现轻微瘙痒，2例患者局部出现炎性丘疹，均不影响治疗。结论：外用他克莫司软膏治疗白癜风疗效较好，不良反应少。     

0.03%他克莫司软膏治疗儿童特应性皮炎的疗效观察

目的:观察外用0.03%他克莫司软膏治疗儿童特应性皮炎的临床疗效和安全性。方法:将60例患者随机分为2组,每组30例,分别对两组患者外用0.03%他克莫司软膏和赋形剂,每天1次,疗程为3周。比较两组疗效。结果:他克莫司软膏组和外用赋形剂的对照组的有效率分别为85.7%和36.7%,两组比较差异有统计学意义(2=12.58,P0.05)。治疗组中7.14%的患者局部有刺激反应,症状于1周后消失。结论:他克莫司软膏用于治疗儿童特应性皮炎疗效明显,个别病例出现局部瘙痒不良反应。     

Interleukin-1beta but not tumor necrosis factor is involved in West Nile virus-induced Langerhans cell migration from the skin in C57BL/6 mice

Langerhans cells are bone marrow-derived epidermal dendritic cells. They migrate out of the epidermis into the lymphatics and travel to the draining lymph nodes where they are responsible for the activation of T cells in the primary immune response. Tumor necrosis factor and interleukin-1beta, have previously been shown to be responsible for Langerhans cell migration in response to contact sensitizers in BALB/C mice; however, which cytokines are responsible for mediating Langerhans cell migration in response to a replicating cutaneously acquired virus such as the West Nile Virus, are not known. We have devised a method for identifying Langerhans cells in the draining lymph nodes using E-cadherin labeling and flow cytometry. We infected tumor necrosis factor-deficient gene knockout mice (tumor necrosis factor-/-) intradermally with West Nile Virus and found that levels of Langerhans cell emigration and accumulation in the draining lymph nodes were similar to wild-type C57BL/6 mice. This was borne out by the finding that high levels of systemic neutralizing anti-tumor necrosis factor antibody failed to inhibit the migration of Langerhans cells from the epidermis and their accumulation in the draining lymph nodes in wild-type C57BL/6 mice. In West Nile Virus-infected, tumor necrosis factor-/- mice treated with systemic neutralizing anti-interleukin-1beta antibodies, however, migration of Langerhans cells from the epidermis and their accumulation in the draining lymph nodes were significantly inhibited compared with control antibody-treated, infected animals. The results indicate that Langerhans cell migration, accumulation in the draining lymph nodes and the initiation of lymph node shut-down in response to a cutaneous West Nile Virus infection is dependent on interleukin-1beta and can occur in the absence of tumor necrosis factor.     

Imiquimod, a topical immune response modifier, induces migration of Langerhans cells

Langerhans cells are bone marrow derived dendritic cells that represent the major antigen-presenting cells in the skin. Langerhans cells take up and process antigen within the epidermis and present processed antigen to T lymphocyte in the regional lymph nodes and thus form an integral part of the cutaneous immune response. The cutaneous immune response can be modified by a number of pharmacologic agents, including corticosteroids, cyclosporine, and retinoids as well as physical agents, such as ultraviolet light. For the most part these agents act by suppressing immune function. A topical immune response modifier, imiquimod has been shown to enhance the cutaneous immune response. Imiquimod has anti-viral and anti-tumor effects in animal models and has been approved for the topical treatment of external genital and perianal warts in humans. The biologic activity of imiquimod in part is due to its effect as a cytokine inducer. Preliminary data suggested that imiquimod could have an effect on Langerhans cells. In order to clarify this effect on Langerhans cells, we examined Langerhans cell morphology and migration in imiquimod-treated skin. The density of Ia + cells decreased 2 d after treatment, falling to approximately 43% by day 10. The Ia positive in cells remaining in the skin appeared larger and more dendritic suggesting an activated state. ATPase staining of epidermal sheet confirmed the decreased number of Langerhans cells. To clarify status of Langerhans cells, the activation of B7 was examined. Activation of B7-1 or B7-2 was not detected. Imiquimod, however, did enhance Langerhans cell migration from skin to draining lymph nodes. This enhanced Langerhans cell migration was also associated with an enhanced allergic contact hypersensitivity. These results suggest that the mechanism of modulation of immune response by imiquimod is in part due to effects on Langerhans cells.     

Antigen presenting cells and T lymphocytes in the induction phase of contact hypersensitivity

When a sensitizing substance that induces contact hypersensitivity, fluorescein isothiocyanate (FITC), was painted on abdominal skin of mice, FITC+ cells appeared in the inguinal lymph node after 24 hours. The FITC+ cell in the lymph node was relatively large in size, and it did not appear to be a T lymphocyte. When FITC was painted on either murine tail skin or skin pre-treated by tape stripping, the number of FITC+ cells in the inguinal lymph node was significantly less than that in the positive control. In the mesenteric lymph nodes, which have a different lymph flow from that of the skin regional lymph node, FITC+ cells did not increase in number, and the few FITC+ cells were not significantly different in number among above-mentioned experimental systems. In the inguinal lymph nodes on the 4th day after painting of picryl chloride (PCl) on the abdominal skin of mice, L3T4+ cells, which expressed an interleukin 2 receptor (IL-2R), increased in number. On the other hand, when PCl was painted on either tail skin or skin treated by tape stripping, L3T4+ IL-2R+ cells did not increase in the skin regional lymph nodes. The number of L3T4+ IL-2R+ cells in the mesenteric lymph nodes did not increase in any of the experimental systems mentioned above. These results suggest some relationship between antigen presenting cells and T lymphocytes, as well as one between the skin and the regional lymph nodes, in an induction phase of sensitization in contact hypersensitivity.     

Langerhans cells migrate to local lymph nodes following cutaneous infection with an arbovirus

Whereas there has been recent interest in interactions between dendritic cells and pathogenic viruses, the role of dendritic cells in the initiation of protective immunity to such organisms has not been elucidated. The aim of this study was to examine whether a resident dendritic cell population in the skin, Langerhans cells, respond to cutaneous viral infections which are effectively cleared by the immune system. We therefore characterized the ability of Langerhans cells to migrate to local draining lymph nodes following infection with the arthropod-borne viruses, West Nile virus or Semliki Forest virus. The data show that major histocompatibility complex class II+/NLDC145+/E-cadherin+ Langerhans cell numbers are increased in the draining lymph nodes of infected mice and this increase is accompanied by a concomitant decrease in the Langerhans cell density in the epidermis. Langerhans cell migration is associated with an accumulation of leukocytes in the lymph node, which is one of the earliest events in the initiation of an immune response. Both the migratory response and the draining lymph node leukocyte accumulation were abrogated if ultraviolet-inactivated instead of live viruses were used, suggesting the activation and subsequent migration of Langerhans cells requires a live, replicating antigen. Our findings are likely to have wider implications for the development of epidermally delivered vaccines and suggest that mobilization of dendritic cells may be involved in the development of immune responses to arthropod-borne viruses.     

Cytokines and chemokines in the initiation and regulation of epidermal Langerhans cell mobilization

Langerhans cells (LC) are members of the wider family of dendritic cells. LC reside in the epidermis where they serve as sentinels of the immune system, their responsibilities being to sample the external environment for changes and challenges and to deliver information (antigen) to responsive T lymphocytes within skin draining lymph nodes. The ability of LC to migrate from the epidermis to regional lymph nodes is therefore of pivotal importance to the induction of cutaneous immune responses. The journey that LC have to make from the skin has a number of requirements. Initially it is necessary that LC disassociate themselves from surrounding keratinocytes and are liberated from other influences that encourage their retention in the epidermis. Subsequently, migrating LC must successfully traverse the basement membrane of the dermal-epidermal junction and make their way, via afferent lymphatics, to draining lymph nodes. Effective entry into lymph nodes is necessary, as is correct positioning of cells within the paracortex. There is increasing evidence that both cytokines and chemokines, and their interaction with appropriate receptors expressed by LC, orchestrate the mobilization and movement of these cells. We here consider the parts played by these molecules, and how collectively they induce and direct LC migration.     

Regulation of the cutaneous allergic reaction by humidity

Humidity is 1 of the environmental factors which regulate skin conditions. Effects of humidity on the cutaneous immune reaction were examined. Contact hypersensitivity to 2,4,6-trinitrochlorobenzene was elicited in C57BL/6 mice. The reaction was greater in mice housed under low humidity conditions (about 10%) for 2 days, at either the induction or elicitation phase, than in mice housed under rather high humidity conditions (80%). After housing under controlled humidity for 2 days, the number of I-A positive cells was 16% higher in the epidermis exposed to the dry condition. The increased population of FITC-positive cells were in regional lymph nodes after painting of FITC during housing under lower humidity. Our study demonstrated that the cutaneous immune reaction is regulated by environmental humidity and suggested 2 possible mechanisms, i.e., increase in Langerhans cells and increased penetration of allergen with low humidity.     

Ultraviolet B radiation suppresses endocytosis, subsequent maturation, and migration activity of langerhans cell-like dendritic cells

Langerhans cells capture exogenous antigens through fluid phase pinocytosis and receptor-mediated endocytosis and migrate to lymph nodes, where they present processed antigen to T cells. Ultraviolet B radiation impairs the antigen-presenting function of Langerhans cells, resulting in antigen-specific immunosuppression of contact hypersensitivity. We tested the notion that ultraviolet B radiation inhibits the endocytic activity of Langerhans cells, leading to impaired migration and maturation. Human monocyte-derived Langerhans cell-like dendritic cells that took up lucifer yellow or fluorescein isothiocyanate dextran exclusively migrated in response to 6Ckine/secondary lymphoid chemokine, and matured, as evidenced by an increase in CD54 and CD86 expression and potent stimulatory activity in allogeneic mixed lymphocyte reaction. Exposing Langerhans cell-like dendritic cells to 20-40 mJ per cm2 of ultraviolet B radiation reduced their endocytic activity in fluid phase pinocytosis (measured by uptake of lucifer yellow) and in receptor-mediated endocytosis (measured by uptake of fluorescein isothiocyanate dextran). Membrane ruffling and CD32 expression were also suppressed by ultraviolet B radiation. Ultraviolet B-irradiated, endocytosing Langerhans cell-like dendritic cells had less movement towards 6Ckine, expressed less CD54 and CD86, and had less effective stimulatory activity in allogeneic mixed lymphocyte reaction than nonirradiated, endocytosing Langerhans cell-like dendritic cells. Endocytosis upregulated tumor necrosis factor alpha production by Langerhans cell-like dendritic cells, but prior ultraviolet B radiation inhibited this enhancement. These data suggested that impaired endocytosis and subsequent inhibitory migration and maturation of Langerhans cells by ultraviolet B radiation could contribute to local immunosuppression of contact hypersensitivity.     

Human epidermal Langerhans cells express the tight junction protein claudin-1 and are present in human genetic claudin-1 deficiency (NISCH syndrome)

Abstract:  Claudin-1 (CLDN1) is a structural tight junction (TJ) protein and is expressed in differentiating keratinocytes and Langerhans cells in the epidermis. Our objective was to identify immunoreactive CLDN1 in human epidermal Langerhans cells and to examine the pattern of epidermal Langerhans cells in genetic human CLDN1 deficiency [neonatal ichthyosis, sclerosing cholangitis (NISCH) syndrome]. Epidermal cells from healthy human skin labelled with CLDN1-specific antibodies were analysed by confocal laser immunofluorescence microscopy and flow cytometry. Skin biopsy sections of two patients with NISCH syndrome were stained with an antibody to CD1a expressed on epidermal Langerhans cells. Epidermal Langerhans cells and a subpopulation of keratinocytes from healthy skin were positive for CLDN1. The gross number and distribution of epidermal Langerhans cells of two patients with molecularly confirmed NISCH syndrome, however, was not grossly altered. Therefore, CLDN1 is unlikely to play a critical role in migration of Langerhans cells (or their precursors) to the epidermis or their positioning within the epidermis. Our findings do not exclude a role of this TJ molecule once Langerhans cells have left the epidermis for draining lymph nodes.     

Stimulation of Langerhans cell migration by tumor necrosis factor alpha (TNF-alpha).

Following topical exposure of mice to skin-sensitizing chemicals, Langerhans cells (LC), many of which bear antigen, are stimulated to migrate via the afferent lymphatics to draining lymph nodes. Consistent with the acquisition of potent immunostimulatory activity, LC while in transit to lymph nodes, are subject to a functional and phenotypic maturation thought to be mediated by granulocyte/macrophage colony-stimulating factor (GM-CSF) and possibly other epidermal cytokines. An interesting question is the nature of the stimulus that initiates the migration of LC from the epidermis. We have examined the influence of intradermal tumor necrosis factor alpha (TNF-alpha), another epidermal cytokine, on the accumulation of dendritic cells (DC) in draining lymph nodes. Murine, but not human, recombinant TNF-alpha caused a rapid and concentration-dependent increase in the frequency of DC in draining nodes. The conclusion drawn is that local production of TNF-alpha provides one signal for LC migration during cutaneous immune and inflammatory responses.     

Visualization and characterization of migratory Langerhans cells in murine skin and lymph nodes by antibodies against Langerin/CD207

Dendritic cells are professional antigen-presenting cells that initiate primary immunity. Migration from sites of antigen uptake to lymphoid organs is crucial for the generation of immune responses. We investigated the migratory pathways specifically of epidermal Langerhans cells by tracing them from the epidermis to the draining lymph nodes. This was possible with a new monoclonal antibody, directed against murine Langerin/CD207, a type II lectin specific for Langerhans cells. In situ, resident, and activated Langerhans cells express Langerin in the epidermis and on their way through dermal lymphatic vessels. Both emigrated and trypsinization-derived Langerhans cells expressed high levels of Langerin intracellularly but reduced it upon prolonged culture periods. Sizeable numbers of Langerin+ cells were found in skin draining lymph nodes but not in mesenteric nodes. Langerin+ cells localized to the T cells areas and rarely to B cell zones. Numbers of Langerin-expressing cells increased after application of a contact sensitizer. In the steady state, Langerhans cells in the skin-draining nodes expressed maturation markers, such as 2A1 and costimulatory molecules CD86 and CD40. These molecules, CD86 and CD40, were further upregulated upon inflammatory stimuli such as contact sensitization. Thus, the novel anti-Langerin monoclonal antibody permits the unequivocal visualization of migratory Langerhans cells in the lymph nodes for the first time and thereby allows to dissect the relative immunogenic or tolerogenic contributions of Langerhans cells and other types of dendritic cells.