慢性荨麻疹患者外周血单一核细胞CC型趋化因子的表达

目的 研究CC型趋化因子家族中调节激活正常T细胞表达和分泌的细胞因子(RANTES)、单核细胞趋化蛋白-1(MCP-1)、单核细胞趋化蛋白-3(MCP-3)和巨噬细胞炎症蛋白(MIP-1α)的mRNA在慢性荨麻疹患者外周血单一核细胞中的表达情况及其相互关系;探讨CC型趋化因子的水平与慢性荨麻疹患者血清总IgE和嗜酸粒细胞阳离子蛋白水平的相关性.方法 逆转录聚合酶链反应(RT-PCR)方法检测31例慢性荨麻疹患者和22例正常人对照外周血单一核细胞中RANTES、MCP-1、MCP-3和MIP-1α的mRNA的表达水平.结果 ①与正常人对照相比,慢性荨麻疹患者外周血单一核细胞中RANTES、MCP-1、MCP-3和MIP-1α的mRNA表达水平均明显增高(P<0.001).②慢性荨麻疹患者中RANTESmRNA的表达水平与MCP-1、MCP-3、MIP-1α的水平呈正相关(P<0.05),MCP-1与MIP-1α的mRNA表达水平之间亦呈正相关(P<0.01),但MCP-1与MCP-3之间、MCP-3与MIP-1α之间的mRNA表达水平无显著相关(P>0.05).③慢性荨麻疹患者中RANTES的mRNA表达水平与血清总IgE和嗜酸粒细胞阳离子蛋白的水平呈负相关(P<0.05);MCP-1、MCP-3和MIP-1α的mRNA表达水平与血清总IgE和嗜酸粒细胞阳离子蛋白的水平之间无显著相关(P>0.05).结论 慢性荨麻疹患者外周血单一核细胞中RANTES、MCP-1、MCP-3和MIP-1α的mRNA表达水平的上调可能与荨麻疹的发病有关.     

Serum chemokine profile in patients with bullous pemphigoid

BACKGROUND: Bullous pemphigoid (BP) is an autoimmune inflammatory disease causing blister formation at the dermoepidermal junction. Cutaneous infiltration of activated CD4+ T cells and eosinophils is an early event in blister formation during the disease process, suggesting that the trafficking of circulating leucocytes through the sites of inflammation is crucial in the pathogenesis of the disease. While the accumulated evidence suggests that some cytokines are involved in the pathogenesis, there have been few reports about serum chemokine profiles in patients with BP. OBJECTIVES: To determine serum profiles of various chemokines and their clinical association in patients with BP. METHODS: Concentrations of 10 chemokines - interferon (IFN)-gamma-inducible protein-10 (IP-10), monokine induced by IFN-gamma (MIG), macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, RANTES, eotaxin, monocyte chemoattractant protein (MCP)-1, MCP-2, MCP-3 and growth-regulated oncogene-alpha- were measured in serum samples from 38 patients with BP, 16 with pemphigus vulgaris (PV) and 17 normal controls using a sandwich immunoassay-based multiplex protein array system. RESULTS: While there was no significant increase in any serum chemokine levels in patients with PV, serum levels of IP-10 and MCP-1 were significantly increased in patients with BP compared with healthy controls. Furthermore, serum levels of IP-10, MIG, MCP-1 and eotaxin in patients with BP increased significantly with disease severity as determined by the area affected. CONCLUSIONS: These observations suggest that an elaborately orchestrated network of chemokines, especially MCP-1 and IP-10, contributes to the pathomechanism of BP.     

The role of chemokines in allergic contact dermatitis

Chemokines are important mediators of immune-mediated skin diseases. Allergic contact dermatitis (ACD) is the most thoroughly investigated T cell-mediated disorder because of the ability to easily reproduce the lesions in humans and the availability of an excellent mouse model. Migration of dendritic cells from the skin to lymph nodes is absolutely required for induction of hapten sensitization, and depends upon expression of CCR7 by mature dendritic cells and SLC in the lymph nodes. During expression of ACD, recruitment of T lymphocytes is driven by chemokines exposed on the surface of endothelial cells or released by activated resident skin cells such as mast cells, fibroblasts and keratinocytes. Chemokines are produced in a coordinated and sequential manner, with IL-8 and RANTES induced by TNF-alpha during early stages, and MCP-1, IP-10, Mig, I-TAC, I-309 and MDC induced by IFN-gamma during later stages. Infiltrating monocytes, dendritic cells and T cells are additional sources of chemokines for further leukocyte accumulation. Distinct T cell subsets express different chemokine receptors, with type 2 cells mostly attracted by eotaxin, MDC, TARC and I-309, and type 1 cells sensitive to IP-10, Mig, I-TAC, RANTES and MIP-1beta. MCP-1 is effective on both subsets. T regulatory cells, which inhibit dendritic cell function and are probably involved in the termination of ACD, are sensitive to MCP-1, MIPs and TARC, but express high levels of CCR8 and are more specifically attracted by I-309. Targeting chemokines and chemokine receptors may offer new opportunities for therapeutic interventions in ACD and other chronic inflammatory skin diseases.     

Serum chemokine profiles in patients with alopecia areata

BACKGROUND: Although chemokines play an important role in various inflammatory diseases, there have been few studies about the role of chemokines in alopecia areata (AA). OBJECTIVES: To determine serum levels of chemokines in patients with AA and their clinical correlations. METHODS: Serum samples from 85 patients with AA, 20 patients with atopic dermatitis, 20 patients with psoriasis vulgaris and 28 normal controls were examined by the cytometric bead array assay assessing monokine induced by interferon (IFN)-gamma (MIG), RANTES, interleukin-8 (IL-8), IFN-inducible protein-10, monocyte chemoattractant protein-1, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta and eotaxin levels. Secreted chemokine levels from peripheral blood mononuclear cells (PBMC) of patients with AA were also investigated. RESULTS: Serum MIG, RANTES, IL-8 and eotaxin levels were selectively increased in patients with AA compared with normal controls. Levels of MIG, RANTES and IL-8 secreted from PBMC of patients with AA were also increased. Furthermore, elevated serum MIG and RANTES levels significantly correlated with the disease activity. RANTES levels were nonsignificantly associated with a predisposition to atopy. CONCLUSIONS: These results suggest that MIG and RANTES play an important role in the development of AA and are useful as markers of disease activity and as therapeutic targets.     

Effects of basophil-priming and stimulating cytokines on histamine release from isolated human skin mast cells

Cell priming and stimulation of different cytokines (which include chemokines and growth factors) are typical features of human basophils. Recently, it has been shown that the macrophage chemotactic protein-1 (MCP-1), RANTES and macrophage inflammatory protein-1α (MIP-1α) are potent direct secretagogues for human basophils and that interleukin-3 (IL-3), IL-5 and granulocyte/macrophage colony-stimulating factor (GM-CSF) are priming factors for subsequent potentiation of mediator release from basophils induced by different stimuli. This observation may be clinically important for the activation and recruitment of inflammatory cells in different immune responses of the skin (e.g. late-phase reactions). The aim of the present study was to investigate whether cytokines and chemokines are also capable of priming or stimulating isolated human skin mast cells (SMC). SMC were either stimulated directly with the cytokines alone or preincubated with these factors for 10 min before being activated with suboptimal concentrations of anti-IgE, A23187 or substance P. IL-3, IL-5, GM-CSF, platelet factor-4 (PF-4), IL-8, MCP-1 and MIP-1α (each at concentrations of 1 ng/ml to 1 μg/ml, log steps) did not significantly modulate histamine release from SMC induced by the three different secretagogues. RANTES exhibited a weak but significant potentiating effect on IgE-mediated activation. Stem cell factor (SCF) as a positive control was able to prime mast cell histamine release strongly. In addition, PF-4, MCP-1, RANTES and MIP-1α were incapable of inducing direct histamine release from SMC. In experiments with isolated human peripheral basophils, however, we observed potent Fc _ε RI-mediated priming effects evoked through IL-3, IL-5, and GM-CSF. We conclude that SMC derived from healthy donors are not targets of (immuno)modulatory factors that prime or stimulate basophils. Received: 21 June 1995     

大疱性类天疱疮患者血清几种趋化因子的检测

目的 探讨大疱性类天疱疮(BP)患者CC型趋化因子家族中嗜酸粒细胞趋化因子(eotaxin)、调节激活正常T细胞表达和分泌的细胞因子(RANTES)、单核细胞趋化蛋白-l(MCP-1)和巨噬细胞炎症蛋白(MIP-lα)的表达.方法 用双抗体夹心ELISA方法检测28例活动期BP患者与30例正常人对照血清eotaxin、RANTES、MCP-1和MIP-lα水平.结果 BP患者血清eotaxin和MCP-1明显高于正常人,P分别<0.001,0.05.RANTES和MIP-lα水平与正常人比较差异无统计学意义(P均>0.05).结论 BP发病中存在趋化因子的异常,eotaxin和MCP-1可能与BP的发病有关.     

趋化因子MCP-1、MIP-lα、RANTES与自身免疫性疾病

趋化因子是一组复杂的小分子量分泌蛋白超家族。自身免疫性疾病的发病机制目前还尚未明了,近年来研究显示在广泛的炎症和自身免疫性疾病中已经观察到上调或下调趋化因子和趋化因子受体的表达,能够影响疾病的易感性、发展和严重性。本文对趋化因子的概况、MCP-1、MIP-lα、RANTES结构与生物学功能以及三者在自身免疫性疾病中的作用作一综述。     

Chemokine release from activated human dermal microvascular endothelial cells – implications for the pathophysiology of scleroderma?

Abstract Long-term exposure to silica (SiO₂) may induce silicosis as well as extrapulmonary diseases such as scleroderma. Infiltration of mononuclear cells and release of proinflammatory cytokines from these cells have been suggested to play a role in the development of inflammatory and immunological events typical of scleroderma as well as of silica-induced scleroderma. We showed that silica is able to directly activate cytokine expression in blood monocytes, collagenase expression in cultured dermal fibroblasts and ICAM-1 expression in human dermal microvascular endothelial cells. In the study reported here we found that silica and TNF· induce mRNA and protein of the chemokines RANTES and MCP-1 in endothelial cells. In addition, we demonstrated that culture supernatants of silica-treated endothelial cells are chemotactic for mononuclear cells from peripheral blood, suggesting that activation of endothelial cells may contribute to the chemotactic gradient necessary for extravasation of inflammatory blood cells into the surrounding tissue found in early scleroderma. However, a polyclonal anti-RANTES antibody failed to block chemotaxis suggesting that other proteins are involved in this phenomenon. We also studied the expression of RANTES in situ in the skin of systemic sclerosis patients and of healthy individuals. We found abundant RANTES mRNA expression in the skin of SSc patients, whereas in control skin no expression was found. From our data we conclude that RANTES and MCP-1 induction by silica may be an initiating event in inflammatory infiltration, whereas TNF·-mediated inflammation may propagate the disease more efficiently. Received: 4 August 1999 / Revised: 17 January 2000 / Accepted: 11 February 2000     

Enhanced production of CC-chemokines (RANTES, MCP-1, MIP-1alpha, MIP-1beta, and eotaxin) in patients with atopic dermatitis

Abstract CC-chemokines are potent molecules that direct the migration of leukocytes to inflammatory foci. To determine their role in inflammation associated with atopic dermatitis (AD), we determined serum levels and spontaneous production of CC-chemokines by peripheral blood mononuclear cells (PBMC) in AD patients using an ELISA. Serum levels of RANTES, MCP-1, MIP-1β, and eotaxin were increased in AD patients (n = 52) compared with normal controls (n = 22). Serum levels of RANTES, MCP-1, and MIP-1β were increased in AD patients with severe disease (n = 19) compared with normal controls (n = 22). Spontaneous production of RANTES, MCP-1, MIP-1α and MIP-1β by PBMC was augmented in AD patients (n = 39) and in patients with severe AD (n = 14) compared with normal controls (n = 20). Serum RANTES levels correlated with total serum IgE levels, eosinophil numbers, and serum lactate dehydrogenase levels. Our results suggest that augmented production of CC-chemokines correlates with inflammation associated with AD. Received: 27 June 2000 / Revised: 25 October 2000 / Accepted: 3 March 2001     

Chemokines and the allergic response

Abstract The β subfamily of chemokines contains-cytokine-like factors which are chemotactic for human basophils and eosinophils. They also stimulate these cells to secrete pro-inflammatory substances such as histamine or eosinophil cationic protein. MCAF/MCP-1, MCP-2, MCP-3, RANTES and MIP-lα all attract and stimulate basophils; MCP-I and MCP-3 are the most potent. RANTES, MCP-3 and to a lesser degree MIP-Ia are chemotactic factors and activators of eosinophils. Cytokines such as IL3, IL5 and GM CSF can augment the responses of these cells to the various chemokines and function as primers. These substances may have particular importance as mediators of allergic inflammation, particularly the late phase component of the response.     

Age-related alterations in the inflammatory response to dermal injury.

Previous studies have documented that the ability to heal wounds declines with age. Although many factors contribute to this age-associated deficit, one variable that has not been carefully examined is leukocyte recruitment and function in wounds. This investigation compares the inflammatory response in excisional wounds of young (age 8 wk) and aged (age 22 mo) mice. In the early inflammatory response, neutrophil content of wounds was similar for both aged and young mice. In contrast, macrophage levels were 56% higher in aged versus young mice (81 +/- 20 vs 52 +/- 13 cells per mm2). In the later inflammatory response, wounds of aged mice exhibited a delay in T cell infiltration, with maximum T cell levels at day 10 in aged mice versus day 7 in young mice. Despite this delay, the eventual peak concentration of T cells was 23% higher in the wounds of aged mice (152 +/- 11 cells per mm2 vs 124 +/- 21cells per mm2). The observed alterations in inflammatory cell content suggested that chemokine production might be altered with age. An elevation of monocyte chemoattractant protein (MCP-1) levels was observed in wounds of aged mice. RNase protection studies, however, revealed that the production of most chemokines, including MIP-2, MIP-1alpha, MIP-1beta, and eotaxin, tended to decline with age. Because optimal wound healing requires both appropriate macrophage infiltration and phagocytic activity, phagocytosis was examined. Compared to young mice, wound macrophages from aged mice exhibited a 37%-43% reduction in phagocytic capacity. Taken together, the data demonstrate age-related shifts in both macrophage and T cell infiltration into wounds, alterations in chemokine content, and a concurrent decline in wound macrophage phagocytic function. These alterations may contribute to the delayed repair response of aging.     

咪唑斯汀联合粉尘螨注射液对慢性荨麻疹患者血清CC型趋化因子水平的影响

目的：研究粉尘螨特异性免疫治疗对慢性荨麻疹患者CC型趋化因子表达水平的影响，探讨特异性免疫治疗的作用机制。方法：将64例粉尘螨过敏的慢性荨麻疹患者随机分为两组，分别采用粉尘螨注射液联合咪唑斯汀治疗或单用咪唑斯汀治疗。应用双抗体夹心酶联免疫吸附试验（ELISA）检测治疗前、后以及正常对照血清中调节激活正常T细胞表达和分泌的细胞因子（RANTES）及单核细胞趋化蛋白-1（MCP-1）的水平。结果：两组慢性荨麻疹患者治疗前血清RANTES和MCP-1的水平均显著高于各自治疗后第16周末（P分别〈0．001和0．05）及正常对照（P均〈0．001）；联合治疗组在治疗后第16周末的RANTES和MCP-1水平以及每周咪唑斯汀用量的评分均显著低于单用咪唑斯汀组（P〈0．01）。结论：RANTES和MCP-1在慢性荨麻疹的发病中起重要作用，特异性免疫治疗对慢性荨麻疹的治疗效果可能与其使RANTES和MCP-1下调的作用有关。     

Expression of CCR2 on monocytes and macrophages in chronically inflamed skin in atopic dermatitis and psoriasis

Monocytes form a significant component of the inflammatory reaction taking place in the skin of atopic dermatitis and psoriasis. Chemokines are pivotal in mediating the attraction of leucocytes to sites of inflammation. The CC-chemokine, monocyte chemotactic protein 1 (MCP-1/CCL2), is expressed by keratinocytes in both atopic dermatitis and psoriasis. MCP-1 binds to the chemokine receptor CCR2 which is known to be expressed on monocytes and macrophages. We examined the expression of CCR2 on peripheral blood monocytes from patients with psoriasis (n=8) and atopic dermatitis (n=7) and found it to be expressed on approximately 90% of the cells, whereas monocytes from healthy donors had a significantly lower CCR2 expression (p<0.05). Skin biopsies from patients suffering from atopic dermatitis and psoriasis revealed that CCR2-positive cells expressed CD163, a marker for monocytes/macrophages. However, not all CD163-positive cells expressed CCR2, which could be interpreted as a mechanism for retaining the macrophages in the skin. Furthermore, we found that keratinocytes are able to express MCP-1, when stimulated with tumour necrosis factor-alpha and/or interferon-gamma in a dose-dependent manner. Thus MCP-1 and CCR2 interaction is likely of importance for the monocyte/macrophage trafficking of inflammatory skin disorders.     

Treatment of recalcitrant pustular psoriasis with infliximab: effective reduction of chemokine expression

Tumour necrosis factor (TNF)-alpha is thought to play a major role in the pathophysiology of psoriasis. Good clinical responses of psoriasis to anti-TNF-alpha-based therapies have recently been demonstrated. We studied the effect of infliximab, a monoclonal antibody against TNF-alpha, on chemokine expression in pustular psoriasis. A 61-year-old man with a 2-year history of severe pustular psoriasis of von Zumbusch type who did not respond to conventional therapies responded rapidly to treatment with infliximab. The clinical response was reflected by an immediate and effective reduction of the neutrophil-attractant chemokines interleukin (IL)-8 and growth-related oncogene (Gro)-alpha as well as of monocyte chemoattractant protein (MCP)-1, as determined by mRNA in situ hybridization of lesional skin. No expression before or after treatment was seen for monokine induced by interferon (IFN)-gamma (MIG) and IFN-inducible protein (IP)-10. Thus, in pustular psoriasis the chemokine expression pattern is dominated by neutrophil-attractant chemokines and MCP-1 while, in contrast to plaque psoriasis, IFN-gamma-inducible lymphocyte-attractant chemokines such as IP-10 and MIG are not abundant. We conclude that anti-TNF-alpha treatment with infliximab is an effective therapy in severe pustular psoriasis which is reflected by downregulation of disease-promoting chemokines such as IL-8, Gro-alpha and MCP-1.     

Angioimmunoblastic lymphadenopathy-type peripheral T-cell lymphoma with cutaneous infiltration: report of a case and its gene expression profile

Angioimmunoblastic T-cell lymphoma is a type of peripheral T-cell lymphoma that is clinically characterized by high fever and generalized lymphadenopathy with or without cutaneous involvement. A 55-year-old Japanese man presented with red papular lesions on the trunk and limbs, oedema, and generalized lymphadenopathy. Histological findings in the lymph nodes showed destructive germinal centres, proliferation of arborizing postcapillary venules, and atypical medium-sized lymphocytes. The cutaneous lesions also contained atypical lymphocytes. Immunohistochemical studies indicated that the neoplastic cells were mature CD4+ T lymphocytes. Southern blot analysis detected a clonal expansion of T-cell receptor beta. Based on these findings, a diagnosis of angioimmunoblastic T-cell lymphoma with cutaneous infiltration was made. Despite systemic chemotherapy, the disease exhibited a high level of activity and continued on a fatal course. An analysis of gene expression profiling using complementary DNA microarrays revealed significant expression of some chemokines and cytokines, e.g. secondary lymphoid tissue chemokine, macrophage inflammatory protein (MIP)-1beta, MIP-3alpha, MIP-3beta, B-lymphocyte chemokine, interleukin-16 and tumour necrosis factor-beta, and an apoptosis-inhibitory protein (FLICE inhibitory protein) in the affected lymph nodes. Profiling of gene expression patterns for a variety of genes in additional cases may be helpful in determining which factors predict the biological and clinical behaviour of angioimmunoblastic T-cell lymphoma or other aggressive malignant lymphomas.     

外阴阴道念珠菌病小鼠模型阴道组织趋化因子MCP-1和MIP-2 mRNA水平测定

目的 探讨趋化因子在白念珠菌性阴道炎模型小鼠阴道组织中的表达情况.方法 采用雌激素化小鼠念珠菌性阴道炎模型,另以雌激素化但未接种白念珠菌组、接种等量白念珠菌但未雌激素化组为对照组,动态观察小鼠阴道灌洗液的真菌生长情况,并在接种后不同时间随机摘取小鼠的阴道组织,半定量RT-PCR方法检测组织内单核细胞趋化蛋白-1(MCP-1)、巨噬细胞炎性蛋白-2(MIP-2)mRNA的表达情况.结果 与未雌激素化的小鼠比较,雌激素化并接种白念珠菌小鼠阴道灌洗液自第2天开始即可发现念珠菌生长并持续到接种后21d,自第4天起阴道组织内有高水平的MCP-1 mRNA表达,并持续高于另外两组直到观察期末.3组小鼠的MIP-2 mRNA表达水平在接种后第2天明显高于完全阴性对照组,但以后各时间点3组间两两比较差异无统计学意义.结论 MCP-1 mRNA高水平表达可能与白念珠菌持续的阴道感染过程有关,而MIP-2在感染过程中的作用不明显.