RANTES在银屑病患者角质形成细胞中的表达及其作用

目的：探讨银屑病皮损局部T淋巴细胞高度浸润的原因，并分析了趋化因子RANTES在此过程中的作用。方法：培养银屑病患者皮损处角质形成细胞，通过微孔小室实验检测其上清液对T淋巴细胞的趋化功能；通过酶联免疫吸附(ELISA)法检测上清液中RANTES的表达。结果：银屑病皮损处角质形成细胞培养上清液对T淋巴细胞的趋化能力明显强于正常对照组；其分泌的RANTES水平也高于正常人。结论：银屑病患者皮损局部T淋巴细胞的大量浸润，部分是由于角质形成细胞具有较强的趋化T淋巴细胞能力，而银屑病角质形成细胞中高表达的RANTES可能是发挥此作用趋化因子之一。     

银屑病患者皮损局部朗格汉斯细胞数量异常机制的研究

目的：探讨银屑病患者皮损局部朗格汉斯细胞(LCs)数量异常的原因。方法：培养银屑病患者皮损处角质形成细胞，通过微孔小室实验检测其上清液对单核细胞的趋化功能；通过酶联免疫吸附法(ELISA)检测上清液中单核细胞趋化蛋白—1(MCP-1)的表达。结果：银屑病患者皮损处角质形成细胞分泌上清液对单核细胞的趋化能力明显强于正常对照组；其分泌的MCP-1水平也高于正常人。结论：银屑病角质形成细胞表达的趋化因子趋化了更多数量的单核细胞至皮损局部，因此，银屑病患者皮损局部LCs的数量理应是增多的。但由于银屑病角质形成细胞表达的其它一些因子也促使了单核细胞衍生的LCs的活化，活化的LCs会迁移至淋巴结或活化后凋亡，又导致其数量减少。因此，银屑病患者皮损局部朗格汉斯细胞数量是一动态变化过程。     

趋化因子与银屑病

趋化因子可分为４个亚家族,不同的趋化因子作用的靶细胞各不相同。趋化因子受体属于Ｇ蛋白偶联受体超家族,与趋化因子共同完成各种生理功能。银屑病患者皮损中单核细胞趋化蛋白－１、白介素－８、α－生长相关癌基因、γ－干扰素诱导的单核因子、γ－干扰素诱导蛋白、活化后可调节的、正常Ｔ细胞表达并可能分泌的因子的表达均与正常人不同,它们趋化各种炎性细胞、Ｔ细胞至皮损局部,白介素－８、α－生长相关癌基因还作用于角质形成细胞使其高度增殖而表现出银屑病的临床及病理特征。针对银屑病患者皮损中趋化因子及其受体的改变,介绍几种抗银屑病药物的新的作用机理。     

趋化因子与银屑病

趋化因子可分为4个亚家族,不同的趋化因子作用的靶细胞各不相同,趋化因子受体属于C蛋白偶联受体超家族,与趋化因子共同完成各种生理功能,银屑病患者皮损中单核细胞趋化蛋白-1、白介素-8、α-生长相关癌基因、γ-干扰素诱导的单核因子、γ-干扰素诱导蛋白、活化后可调节的、正常T细胞表达并可能分泌的因子角质形成细胞使基伉度增殖而表现出银屑病的临床及病理特征,针对银屑病患者皮损中趋化因子及其受体的改变,介绍几种抗银屑病药物的新的作用机理。     

银屑病患者角质形成细胞对朗格汉斯细胞免疫刺激功能的影响

为探讨银屑病患者皮损处角质形成细胞对朗格汉斯细胞（LCs）免疫刺激功能的影响，从健康者外周血分离单核细胞，经IL－4＋GM－CSF＋TGF－β1培养5天后分化发育为1Cs,再加入银屑病患者角质形成细胞培养上清继续培养2天，然后通过Leica显微镜观察其形态；通过混合淋巴细胞反应分析其种T的刺激功能。结果显示，以健康人角质形成细胞上清为对照，LCs经银屑病患者角质形成细胞上清培养2天后，拥有不规则突起的细胞明显较正常组增多。同时，其刺激T淋巴细胞增殖的能力也增强。结果表明，银屑病患者角质形成细胞上清培养的LCs表现出较强的免疫刺激功能，并在银屑病相关的免疫反应过程中发挥了重要作用，提示这可能与银屑病患者角质形成细胞表达的某些因子有关。     

角质形成细胞生长因子及神经细胞生长因子在银屑病患者皮损中的表达

目的 探讨角质形成细胞生长因子(KGF)及神经细胞生长因子(NGF)与银屑病的关系及银屑病皮损中间质细胞与角质形成细胞的相互作用。方法 应用免疫组化技术分别检测33例银屑病患者皮损、8例非皮损和13例正常人皮肤中KGF、KGF受体、NGF、NGF受体的表达。结果 KGF、KGF受体在银屑病患者基底层和棘细胞下层有极强的表达,与非皮损组及正常对照组间差异有显着性(P<0.05),而非皮损组与正常对照组间差异无显着性(P>0.05).KGF、KGF受体在真皮层未见表达。NGF、NGF受体则主要表达在颗粒层和棘细胞上层,皮损组与非皮损组之间及非皮损组与正常组之间均差异有显着性(P<0.05)。结论 银屑病患者皮损中KGF及NGF可能介导了角质形成细胞的增殖与分化不全;银屑病皮损中存在着间质细胞和角质形成细胞相互作用的紊乱。     

神经肽CGRP对银屑病单核细胞趋化功能的调节

为探讨神经肽对银屑病免疫细胞的调节，及其在银屑病神经免疫发病机制中的作用，本研究利用体外细胞培养技术分离培养单核细胞，分别加入外源性神经肽降钙素基因相关肽（calcitonin gene-related peptide,CGRP)及其受体拮抗剂CGRP8－37。用ELISA检测培养单核细胞上清液中趋化因子的含量；利用微型趋化小室，观察CGRP对单核细胞趋化活性的调节。结果CGRP诱导银屑病活化的单核细胞分泌趋化因子巨噬细胞炎性蛋白－1α(macrophage inflammatory protein-1α，MIP-1α）和单核细胞趋化性蛋白－1α（monocyte chemotactic protein-1α,MCP-1α)增加，受体拮抗剂CGRP8－37则抑制这种诱导作用，同时CGRP促进单核细胞对淋巴细胞和中性粒细胞的趋化活性，用CGRP8－37后则趋化活性减弱。提示银屑病皮损内神经肽CGRP可以通过受体诱导单核巨噬细胞分泌MIP－1α和MCP－1α趋化因子，使淋巴细胞和中性料细胞在局部皮损区定向迁移与聚集，促进局部炎性细胞的浸润。     

应用RT—PCR技术检测银屑病皮损中IGF—1受体mRNA

为了了解银屑病皮损中ＩＧＦ－１受体ｍＲＮＡ的表达,采用ＲＴ－ＰＣＲ对２０例银屑病和正常人皮肤组织块进行检测,结果证实银屑病患者表皮中ＩＧＦ－１受体ｍＲＮＡ表达显著高于对照（Ｐ〈０．０１）,真皮中未有ＰＧＦ－１受体与银屑病角度形成细胞在转录水平与增殖,分化有关,ＩＧＦ－１通过旁分泌机制作用于角质形成细胞上受体。     

葡萄球菌性超抗原对银屑病患者外周血淋巴细胞产生白介素-2、γ-干扰素的影响

银屑病患者外周血淋巴细胞对葡萄球菌性超抗原（葡萄球菌肠毒素B，SEB)和链球菌超抗原刺激反应增强，且经SEB刺激的银屑病患者外周血淋巴细胞(PBLC)培养上清液可促进角质形成细胞增殖，表达HLA-DR和Fas抗原，继而诱导细胞凋亡。为了进一步探讨培养上清液对角质形成细胞的作用机制，笔者检测了SEB刺激前后的银屑病患者淋巴细胞培养上清液中白介素(IL）-2、γ-干扰素（IFN-γ)的水平变化，现将结果报告如下。     

寻常性银悄病患者外周血淋巴细胞中红细胞趋化因子受体的表达

银屑病的发病机制尚不清楚，在银屑病患者体内存在着多种体液和细胞免疫紊乱。研究表明调节活化和正常T细胞表达及分泌的趋化因子（RANTES）、单核细胞趋化蛋白-1（MCP-1）和白介素8（IL-8）等趋化因子在银屑病患者局部皮损表达明显增强，它们在银屑病的发病中起重要作用。红细胞趋化因子受体（ECKR）即红细胞表面的达菲抗原/趋化因子受体（Duffy antigen/receptor for chemokines,DARC）,     

Psoriatic lesional keratinocytes promote the maturation of human monocyte-derived Langerhans cells

BACKGROUND: Langerhans cells (LCs) are important antigen-presenting cells in the epidermis and may play a key role in the pathogenesis of psoriasis. It has been proven that LCs isolated from psoriatic lesions are abnormal. However, the mechanism of the abnormality has not been reported so far. OBJECTIVE: In the present study, we investigated the effect of psoriatic lesional keratinocytes on the maturation of LCs. METHODS: Monocytes isolated from healthy peripheral blood could differentiate into LCs in the presence of granulocyte-macrophage colony-stimulating factor, interleukin (IL) 4 and transforming growth factor beta1 for 5 days. Then, human monocyte-derived LCs were cultured with supernatants from psoriatic lesional keratinocytes for another 2 days. Their phenotypes and phagocytic capacity were analyzed by flow cytometry. IL-12 secreted by LCs was determined by ELISA. RESULTS: Supernatants from psoriatic lesional keratinocytes could up-regulate the expression of HLA-DR and CD86 on LCs more significantly than supernatants from healthy keratinocytes, but less powerfully than lipopolysaccharide. The levels of IL-12 secreted by LCs also increased. In contrast, the expression of CD1a on LCs and their phagocytic capacity were reduced. CONCLUSION: Human monocyte-derived LCs cultured with supernatants from psoriatic lesional keratinocytes displayed the characteristics of maturation. This suggests that psoriatic lesional keratinocytes might secrete some factors that could promote the maturation of LCs, which may play important roles in immune reactions related to psoriasis.     

The inhibitory effect of VitD3 on proliferation of keratinocyte cell line HACAT is mediated by down-regulation of CXCR2 expression

Psoriasis is a disease characterized by inflammation and increased population of hyperproliferative keratinocytes. It is well known that chemokines and chemokine receptors, such as interleukin-8 and its receptors (CXCR1 and CXCR2), play important roles in the pathogenesis of psoriasis. So far, examination of CXCR2 expression in psoriatic lesional keratinocytes by FACS calibur has not been reported and whether VitD3 inhibits psoriatic lesional keratinocyte proliferation through down-regulation of CXCR2 expression has not been elucidated. In the present study, CXCR2 expression in psoriatic lesional keratinocytes and HACAT treated with VitD3 was detected by flow cytometry. The proliferative capacity of HACAT treated with VitD3 was assayed by MTT assay. The results showed that CXCR2 expression in psoriatic lesional keratinocytes was higher than that in normal human keratinocytes. At the correct concentration VitD3 could inhibit human keratinocyte proliferation and down-regulate CXCR2 expression in HACAT. The data demonstrate that the inhibitory effect of VitD3 on keratinocyte proliferation might be mediated by down-regulation of CXCR2 expression.     

银屑病患者角质形成细胞培养上清液对单核细胞形态、表型的影响

目的　探讨银屑病角质形成细胞 (keratinocytes ,KC)在单核细胞向郎格汉斯细胞 (langerhanscells ,LC)分化过程中的作用。方法　从健康志愿者外周血分离单核细胞 ,分别经银屑病患者及正常人KC培养上清单独培养 5天 ,然后通过Leica显微镜观察其形态 ,通过流式细胞仪分析其表型。结果　健康志愿者外周血单核细胞经银屑病患者KC上清培养后 ,细胞的聚集体明显较正常组减少 ,而与阴性对照组间无显著差异 ;CDla、CD80、CD86的表达均较正常人KC上清组减弱。结论　银屑病患者KC中缺乏某些因子而影响了单核细胞向LC的分化 ,或表达了某些因子而抑制了单核细胞向LC的分化。     

UM4D4+ (CDw60) T cells are compartmentalized into psoriatic skin and release lymphokines that induce a keratinocyte phenotype expressed in psoriatic lesions

UM4D4 (CDw60), the surface molecule of a novel antigen-independent T-cell activation pathway, was found to be highly expressed on lesional psoriatic T cells. To examine whether UM4D4 represents a T-cell activation pathway for psoriatic T cells, a T-cell line was initiated from an acute skin lesion and cloned by limiting dilution. Clonality was verified by analysis of T-cell receptor gene rearrangement. All T-cell clones tested, whether CD4+2H4+CD8-, CD4+2H4-CD8-, or CD4-CD8+CD11b-, expressed UM4D4 and were activated by the monoclonal antibody anti-UM4D4. Lesional psoriatic T-cell clones were heterogeneous in the degree of anti-UM4D4-induced proliferation and in their production of IL-2 and gamma-interferon. Lymphokines released by anti-UM4D4 activation were capable of inducing ICAM-1 and HLA-DR expression on cultured normal keratinocytes. Thus, the high expression of UM4D4 on T-cells in psoriatic skin provides an alternative mechanism for T-cell activation that may be operative in the psoriatic lesional milieu. Indeed, activation of lesional T-cells through the UM4D4 molecule resulted in release of lymphokines that directly induced keratinocytes to express a phenotype displayed in psoriatic skin lesions.     

Mast cell density and IL-8 expression in nonlesional and lesional psoriatic skin

BACKGROUND: An important cellular aberration at sites of psoriatic inflammation is an increase in the number of dermal mast cells. Being multifactorial immune effector cells, it is believed that mast cells play an essential role in perpetuating the inflammatory process of psoriasis. However, factors responsible for the infiltration and accumulation of mast cells in psoriatic lesions are largely unknown. Recent studies have demonstrated that Interleukin-8 (IL-8) exerts strong chemotactic effects on mast cells in vitro. Overexpression of IL-8 has also been reported in psoriatic lesions. In this study, we have found a correlation between the expression of IL-8 and dermal mast cell density in lesional psoriatic skin as compared to nonlesional psoriatic skin. METHODS: Four-mm punch biopsies were taken from 14 psoriatic patients and eight healthy volunteers. Using immunohistochemical techniques, 8 microm sections of lesional psoriatic, nonlesional psoriatic, and normal control samples were evaluated for dermal mast cell density and the density of IL-8 expressing keratinocytes. RESULTS: It was found that dermal mast cell density in lesional psoriatic, nonlesional psoriatic, and normal skin was 105.4 +/- 71.2, 42.3 +/- 30.1, and 47.5 +/- 32.5 mast cells/mm(2), respectively. IL-8+ keratinocyte density in lesional psoriatic, non lesional psoriatic, and normal skin was 171.5 +/- 67.1, 25.4 +/- 14.9 and 20.6 +/- 8.7 IL-8+ Keratinocytes/mm(2), respectively. CONCLUSIONS: The results of this study suggest that increased levels of IL-8 in the keratinocytes of psoriatic plaques play a contributing role in the migration of mast cells to lesion sites.     

维生素D_3抑制角质形成细胞MCP-1的表达

目的　观察维生素D3 对角质形成细胞单核细胞化学吸引蛋白质 1(monocytechemoattractantprotein 1,MCP 1)的表达影响 ,探讨其在治疗银屑病中的作用机制。方法　培养角质形成细胞株HACAT ,并用不同浓度的维生素D3 处理 2 4h ,通过酶联免疫吸附 (ELISA)法检测上清液中MCP 1的表达 ;通过RT PCR法进一步检测细胞中MCP 1mRNA的水平。结果　经过浓度为 7.8× 10 -12 mol/L～ 6.2 5× 10 -11mol/L的维生素D3 处理过的角质形成细胞株HA CAT ,其分泌至上清中的MCP 1水平明显受到抑制 ,细胞中MCP 1mRNA的水平低于正常对照组。结论　维生素D3 在治疗银屑病过程中很有可能是通过抑制了角质形成细胞中MCP 1的表达 ,从而减弱了银屑病患者角质形成细胞趋化单核细胞及巨噬细胞的能力 ,阻止了银屑病皮损局部的炎症反应。