不同预后尖锐湿疣组织中Toll样受体3、9的表达

目的 探讨不同预后尖锐湿疣（CA）患者皮损局部Toll样受体3（TLR3）和Toll样受体9（TLR9）表达定位及TLR3 mRNA、TLR9 mRNA的表达情况。方法 免疫组化SP法及反转录聚合酶链反应（RT-PCR）法检测10例CA复发者、14例CA无复发者及10例包皮组织中TLR3 及TLR9表达情况。结果 TLR3、TLR9在无复发CA皮损中表达以棘层、颗粒层为主;在复发组CA皮损中表达以基底层、棘层为主。无复发组CA组织中TLR3mRNA表达较对照组及复发组均明显升高（P ＜ 0.01、P ＜ 0.05）,TLR9 mRNA较对照组明显升高（P ＜ 0.05）,较复发组差异无统计学意义。结论 表皮TLR3 及TLR9的表达部位和表达量的改变可能与CA预后有关,TLR3的影响可能更大。     

Detection of migration inhibitory factor (MIF) by a monoclonal antibody in the microvasculature of inflamed skin

Summary Macrophage migration inhibitory factor (MIF) is known as a mediator of cellular immunity with specific effects on the differentiation of mononuclear phagocytes. There is little information on the production of MIF in vivo and its role in the pathophysiology of inflammation. We studied the distribution of MIF in various tissues with a monoclonal antibody against human MIF (1C5/B) using the indirect immunoperoxidase method. Here, we investigate the expression of MIF on endothelial cells of dermal vessels. Our results show that dermal vessels may constitutively express MIF and can be strongly activated to express MIF in acute inflammations such as eczema and psoriasis in contrast to the chronically infiltrated skin from patients with pseudolymphomas and sarcoidosis. In these cases a possibly MIF defective state of vessels and a restriction of positive vessels to distinct anatomical sites of the inflamed skin was detected. The significance of the described association of MIF with vascular endothelium is still a matter of speculation. MIF expression on endothelium may provide an important differentiogenic signal for mononculear phagocytes on their way to the tissue site.     

Preferential expression of alphaEbeta7 integrin (CD103) on CD8+ T cells in the psoriatic epidermis: regulation by interleukins 4 and 12 and transforming growth factor-beta

BACKGROUND: Intraepidermal T lymphocytes are a critical element for sustaining the lesional pathology of psoriasis. Integrin alphaEbeta7 (CD103), a ligand for E-cadherin, may play a role in the localization of pathogenic T cells within the epidermis of psoriatic lesions. However, little information is available regarding alphaEbeta7 expression on intraepidermal T cells in psoriasis. OBJECTIVES: To examine alphaEbeta7 expression on intraepidermal T cells in psoriatic lesions and the regulation of alphaEbeta7 expression on T cells in response to cytokines. METHODS: T-cell expression of alphaEbeta7 was examined by immunohistochemistry and flow cytometry. In vitro regulation of alphaEbeta7 expression on CD4+ or CD8+ T cells purified from peripheral blood of healthy donors was also examined. RESULTS: Immunohistochemical staining revealed expression of alphaEbeta7 on a greater proportion of epidermal T cells than dermal T cells. Nearly 30% of intraepidermal CD4+ T cells were found to express alphaEbeta7 on flow cytometry, whereas more than 80% of intraepidermal CD8+ T cells expressed this integrin. In contrast, few T cells expressed alphaEbeta7 in the peripheral blood of psoriatic patients. The in vitro culture experiment confirmed that alphaEbeta7 was preferentially expressed on CD8+ T cells after stimulation with anti-CD3 monoclonal antibodies. Addition of transforming growth factor-beta and interleukin-4 upregulated alphaEbeta7 expression on T cells, whereas interleukin 12 downregulated this. Furthermore, alphaEbeta7 expression on established memory CD8+ T cells was not so reversible as that on CD4+ T cells. CONCLUSIONS: Preferential and stable expression of alphaEbeta7 on CD8+ T cells may be involved in the lesional pathology of psoriasis.     

The severity of irritant contact dermatitis in various strains of mice correlates with endothelial expression of migration inhibitory factor (MIF)

Summary Irritant contact dermatitis to croton oil in BALB/cByJ, C57Bl/6J and six recombinant inbred CxB strains of mice was investigated in relation to variations in endothelial migration inhibitory factor (MIF) reactivity. MIF has been shown to be a mediator of cellular immunity and operates as a differentiation signal inducing an inflammatory type of macrophage. The intensity of the ear swelling response reached a maximum 8 h after induction of contact dermatitis, with highest values in BALB/cByJ and CxB4/ByAH mice and weak reactions in CxB2/ByAE, CxB7/ByAK, C57Bl/6J and CxB1/ByAD mice. After the same time period (8 h) cryostat sections were immunostained for capillary endothelium expressing MIF. The most pronounced MIF expression was observed in BALB/ cByJ mice, and CxB4/ByAH mice showed intermediate reactions and the other strains weak reactions. Endothelial MIF expression correlated well with the intensity of ear swelling (Pearson's correlation coefficient 0.82). Patterns of endothelial MIF expression in recombinant inbred strains suggest that endothelial MIF expression is not under the control of a single gene. Our data support the hypothesis that endothelial MIF expression plays a prominent role in inflammatory events and correlates with the severity of inflammation.     

Reduction of inflammatory slan (6‐sulfo LacNAc) dendritic cells in psoriatic skin of patients treated with etanercept

Dermal dendritic cells (DCs) play a central role in the immunopathology of psoriasis. We previously identified slanDCs as pro‐inflammatory TNF‐α, IL‐23‐ and IL‐12‐producing DCs in human blood and as prominent inflammatory dermal TNF‐α secreting and CD11c‐positive DC subset in psoriasis. Here, we ask for the effects of TNF‐α‐inhibition on inflammatory slanDCs in skin and blood of 10 patients with psoriasis during 24 weeks of treatment with etanercept. Treatment with etanercept reduced the frequency of dermal slanDCs but did not induce apoptosis as determined by lack of increased active caspase‐3‐expression. In parallel, we found increased frequencies of slanDCs in blood which expressed lower levels of HLA‐DR. Stimulating slanDCs isolated from the blood of healthy donors in vitro induced a strong production of IL‐1β, IL‐6, IL‐23 and IL‐12p70. This capacity was efficiently reduced in the presence of etanercept, thereby indicating that TNF‐α is an autocrine stimulus for maturation and pro‐inflammatory cytokine production of slanDCs. In vivo, we noticed that treatment with etanercept did reduce the number of dermal slanDCs in parallel to the overall expression of TNF‐α and IL‐23p19. However, successful treatment did not down‐regulated the percentage of dermal slanDCs that stained positive for TNF‐α and IL‐23p19 indicating that remaining slanDCs kept their pro‐inflammatory capacity. This study provides novel insights into the immune regulatory properties of etanercept at the level of inflammatory slanDCs in vivo in skin and blood as well as in vitro.     

Elevation of circulating neutrophil extracellular traps,interleukin (IL)-8, IL-22, and vascular endothelial growth factor in patients with venomous snake mamushi (Gloydius blomhoffii) bites

Mamushi bites cause swelling and pain that extend from the bitten site. The coagulopathic, anti-coagulopathic, and vasculopathic actions of mamushi venom result in various laboratory abnormalities, occasionally with muscular, renal, and other organ damage. We investigated the serum biomarkers that were associated with the pathogenesis of mamushi bites, focusing on markers related to tissue-damage and neutrophil activation. Twenty patients (one case of grade 2, 13 cases of grade 3, and six cases of grade 4 of severity) seen by us in one summer season were enrolled. Peripheral blood samples were taken from the patients on day 0, day 2, and day 7 after mamushi bites. In addition to routine blood examination, serum samples were subjected to enzyme-linked immunosorbent assay for citrullinated histone H3 (CitH3), interleukin (IL)-8, IL-17A, IL-22, vascular endothelial growth factor (VEGF), high mobility group box protein 1 (HMGB1), tumor necrosis factor (TNF)-α, and IL-33. Creatinine kinase (CK) values significantly correlated with prothrombin time (PT) levels, suggesting that muscular damage is associated with exaggerated coagulation and fibrinolysis. In the vast majority of patients, HMGB1, TNF-α, and IL-33 were under detection levels. Neutrophil counts did not correlate with PT or CK, indicating that the coagulation disorder and muscular damage were virtually independent of the neutrophil activation. The neutrophil number significantly correlated with CitH3, a representative marker of neutrophil extracellular traps. Moreover, there were significant correlations between neutrophil number, CitH3, IL-8, IL-22, and VEGF. Our study suggests that there are two major cascades in mamushi bites. One is an already characterized venom effect on coagulation, vessels, and muscles. In the other novel cascade, we propose that neutrophil activation with IL-8 leads to the production of IL-22 and VEGF. This sequential event may contribute to both vascular damage and repair.     

Macrophage migration inhibitory factor (MIF) in the stratum corneum: a marker of the local severity of atopic dermatitis

Different biomarkers are used to evaluate the severity of atopic dermatitis (AD); however, it remains difficult to determine the severity of localized skin lesions. MIF plays an essential role in the pathophysiology of skin inflammation. To establish whether the MIF level in the stratum corneum (SC) serves as a marker of the severity of AD lesions, we examined the SC MIF (scMIF) levels in AD patients. The SC of the cheek, neck and upper arm skin was collected using tape stripping, and the scMIF levels were measured. Consequently, the scMIF levels were found to be significantly higher in the involved skin lesions than the uninvolved areas within the same patient. Moreover, the scMIF levels were significantly correlated with the severity of local skin lesions. These findings suggest that the scMIF level can be used as an effective marker for evaluating the local severity of AD.     

丹参对急性胰腺炎患者血栓素A2／前列腺环素平衡和白介素-6、白介素-8水平的影响

目的探讨急性胰腺炎（AP）患者血栓素A2/前列腺环素（TXA2/PGI2）平衡和白细胞介素-6（IL-6）、IL-8的变化情况,及丹参治疗急性胰腺炎的作用机制。方法50例AP住院AP患者,包括重症急性胰腺炎（SAP）组13例、轻症急性胰腺炎（MAP）组37例,比较组间TXA2/PGI2平衡和IL-6、IL-8水平的差异。50例患者给予禁食、胃肠减压、生长抑素、质子泵抑制剂、抗感染及营养支持等治疗,并随机分为丹参治疗组（A组）,在上述治疗基础上给予丹参注射液250ml,静脉点滴2次／d,7d为一个疗程）和一般治疗组（B组）。同时取健康体检者20例作为对照组（C组）。治疗前后取外周静脉血标本,测定TXA2、PGI2、IL-6、IL-8的含量。结果治疗前与C组比较,SAP组、MAP组患者外周血TXA：含量明显升高,PGI2含量降低,IL-6、IL-8含量升高（均P〈0．05）。与MAP组比较,SAP组TXA2、PGI2、IL-6、IL-8含量的异常变化更明显。治疗7d后,A组TXA2、PGI2含量与治疗前比较,已基本恢复正常（均P〈0．05）,B组TXA2、PGI2含量与治疗前比较,无明显变化（均P〉0．05）;治疗7d后,A组、B组IL-6、IL-8含量均显著低于治疗前（均P〈0．05）,与B组比较,A组下降更加明显（均P〈0．05）。结论AP患者存在TXA2/PGI2平衡紊乱和IL-6、IL-8异常升高现象。丹参治疗AP的机制可能是通过纠正血管活性物质分泌失衡,使血浆TXA2含量下降,PGI2含量上升,使TXA2/PGI2比值失衡恢复,并且抑制体内IL-6、IL-8的异常分泌,减轻AP患者全身炎症反应的水平,改善微循环障碍,最终使AP患者的病程得以逆转。     

白芍总苷对角质形成细胞增殖及血管内皮生长因子和白介素-23表达的影响

目的 研究白芍总苷（TGP）对角质形成细胞增殖和分泌血管内皮生长因子（VEGF）和白介素（IL）-23的影响,探讨可能涉及的信号转导通路。方法 不同浓度TGP作用于体外培养的HaCaT细胞株,噻唑蓝（MTT）法观察TGP对HaCaT细胞增殖活性的影响。将HaCaT细胞分为三组,即对照组不加任何刺激因素,TGP组分别加入6种不同浓度的TGP,SB203580组在加入10 mol/L SB203580预处理2 h后加入125 mg/L TGP。实时定量PCR（RT-PCR）方法和ELISA方法检测TGP对HaCaT细胞VEGF和IL-23表达的影响;免疫印迹技术观察TGP作用于HaCaT细胞后p38的磷酸化及SB203580对p38磷酸化的影响。结果 TGP在低浓度（0.5、2.5 mg/L）时对HaCaT细胞增殖有促进作用,浓度≥12.5 mg/L时反而对细胞的增殖有抑制作用,至125 mg/L时抑制作用最强。TGP在低浓度（0.5、2.5 mg/L）时对HaCaT细胞VEGF和IL-23 mRNA和蛋白的表达有促进作用,12.5 ～ 125 mg/L时内可抑制HaCaT细胞VEGF mRNA和蛋白的表达,62.5 ～ 125 mg/L时可抑制IL-23 mRNA和蛋白的表达。TGP可时间依赖性地诱导HaCaT细胞p38的磷酸化,磷酸化p38 蛋白于125 mg/L TGP作用5 min后达到高峰,表达水平为0.3314 ± 0.0245,10 min后减弱至0.2173 ± 0.0189,但均高于对照组水平;30 min后表达水平降为0.1664 ± 0.0201;SB203580可减弱其作用,SB203580预处理组磷酸化p-p38 表达水平为0.1529 ± 0.0147。结论 TGP可抑制HaCaT细胞的增殖及VEGF和IL-23 mRNA和蛋白的表达,p38MAPK信号途径可能介导其抑制作用。     

Efficacy and safety of ixekizumab treatment in Japanese patients with moderate‐to‐severe plaque psoriasis: Subgroup analysis of a placebo‐controlled,phase 3 study (UNCOVER‐1)

The present study describes a subgroup analysis of 33 Japanese patients participating in UNCOVER‐1, an international, placebo‐controlled, phase 3 study of ixekizumab in patients with moderate‐to‐severe psoriasis. Patients were randomized to a placebo (n = 13) or ixekizumab 80 mg every 4 (IXEQ4W, n = 12) or 2 (IXEQ2W, n = 8) weeks, from week 0–12. At week 12, ixekizumab‐treated patients with a static Physician Global Assessment score 0 or 1 (sPGA [0,1]; n = 16) were re‐randomized to a placebo (n = 6), ixekizumab 80 mg every 12 (IXEQ12W, n = 5) or 4 (IXEQ4W, n = 5) weeks, from week 12–60. At week 12, more ixekizumab‐treated versus placebo‐treated patients achieved sPGA (0,1) (≥66.7% vs 0%), ≥75% improvement in Psoriasis Area and Severity Index (≥75% vs 0%), and sPGA (0) or 100% improvement in Psoriasis Area and Severity Index (both ≥33.3% vs 0%), with improved symptoms and quality of life. At week 60, 100% (IXEQ4W), 40.0% (IXEQ12W) and 16.7% (placebo) had maintained sPGA (0,1). From week 0–12, treatment‐emergent adverse events were 76.9% (placebo), 75.0% (IXEQ4W) and 87.5% (IXEQ2W), and from week 12–60 were 66.7% (placebo) and 100% (IXEQ12W, IXEQ4W). Ixekizumab‐treated patients had no severe treatment‐emergent adverse events, and one serious TEAE (IXEQ4W); infection was the most frequent treatment‐emergent adverse event. In conclusion, ixekizumab for 60 weeks was effective and safe for Japanese patients with moderate‐to‐severe psoriasis, in line with the overall findings from UNCOVER‐1.     

Increased frequency of intracellular interleukin (IL)-13 and IL-10, but not IL-4, expressing CD4+ and CD8+ peripheral T cells of patients with atopic dermatitis

BACKGROUND: A number of studies exist demonstrating the increased expression of type 2 cytokines and decreased capacity to produce interferon-gamma (IFN-gamma) in peripheral blood mononuclear cells (PBMCs) of patients with atopic dermatitis (AD). OBJECTIVES: To clarify the results of recent studies concerning the role of interleukin (IL)-4 and IL-13 in PBMCs of AD patients, we analysed the activation status of lymphocyte subpopulations. METHODS: We measured the intracellular expression and serum levels of certain type 1 and type 2 cytokines, using cell surface and intracellular cytokine staining, flow cytometry and enzyme-linked immunosorbent assay techniques. RESULTS: The frequency of IL-10 and IL-13 producing CD4+ and CD8+ T cells was significantly higher in patients with AD, while the frequency of IFN-gamma secreting helper and cytotoxic T cells was significantly lower in patients with AD than in control subjects. The serum levels of IL-10 and IL-13 were also significantly increased. There were no significant differences observed between the experimental groups in the frequency of IL-4 producing CD4+ and CD8+ cells. CONCLUSIONS: This study demonstrates a type 2 cytokine production in the CD4+ and CD8+ T cells of AD patients, which is characterized by an elevated IL-13, but not by IL-4 secretion, and by an increased level of the immunoregulatory IL-10, which can contribute to a decrease in IFN-gamma expression.