Cytokine gene polymorphisms in psoriasis

BACKGROUND: Cytokine production is under genetic control, and certain allelic variants of cytokine genes are associated with higher or lower cytokine production in vitro and in vivo. Psoriasis is associated with an overexpression in the involved skin of T-helper cell type 1 (Th1) cytokines, e.g. interferon (IFN) -gamma and tumour necrosis factor (TNF) alpha and relative underexpression of Th2 cytokines, e.g. interleukin (IL) -4 and IL-10. Objective We investigated the hypothesis that allelic variants of genes for a high production of Th1 cytokines or TNF-alpha, or conversely low production of Th2 cytokines might represent a risk factor for developing psoriasis. METHODS: Genotyping for IFN-gamma, IL-10, IL-4 and TNF-alpha was undertaken for 84 patients with psoriasis and compared with control data on file. RESULTS: Genotype frequencies showed no differences between patients and controls for IFN-gamma, TNF-alpha or IL-4. For IL-10, patients with late onset psoriasis (over 40 years) were more likely to be heterozygous at position - 1082 (P = 0.02), corresponding to intermediate production of IL-10 in vitro and in vivo. CONCLUSIONS: Psoriasis is not determined by a genotype consistent with high production of Th1 cytokines or low production of Th2 cytokines. Thus, the Th1 cytokine profile found in psoriatic plaques is most likely a consequence of local factors.     

寻常型银屑病患者几种血清细胞因子的检测

目的:探讨血清中白介素12(IL-12)、IL-18、IL-4及IL-10水平与寻常型银屑病的发病关系。方法:采用双抗体夹心ELISA法检测30例寻常型银屑病患者及15例健康志愿者血清IL-12、IL-18、IL-4及IL-10的水平。结果:银屑病组IL-18水平显著增高,IL-10水平显著降低,与健康对照组相比差异均有统计学意义(P<0.05)。结论:IL-18、IL-10在银屑病的发病中可能发挥重要作用。     

Epidermal CD8+ T cells in chronic plaque psoriasis are Tc1 cells producing heterogeneous levels of interferon-gamma

The majority of epidermal CD8+ T cells in chronic plaque psoriasis are activated Tc1 cells producing interferon-gamma and no interleukin-4, a small proportion of which express NK-T receptors. To quantitate their level of cytokine production and characterize them further, CD8+ T cells were isolated from epidermal cell suspensions of lesional biopsies from 24 patients with chronic plaque psoriasis. T-cell lines (TCL) were established by culture of CD8+ T cells with feeders and IL-2 for 11 days and expansion with PHA. Ten TCL were stained for surface markers; 6 were cloned with PHA by limiting dilution. Interferon-gamma, interleukin-4 and interleukin-10 production was measured by ELISA after PMA/anti-CD3 activation of 15 TCL and 39 CD8+ T-cell clones. The 10 TCL stained were CD8alphabeta+ (93.3%), T-cell receptor-alphabeta+ (99.5%), costimulatory molecule CD28+ (90.1%), with a small CD8alphaalpha+ population (2.3%). No NK-T-cell receptor CD158a or CD158b expression was detected, whilst CD94 was expressed on 6.2% of cells in 6/9 TCL. All the TCL and 37/39 CD8+ T-cell clones produced interferon-gamma but no or minimal interleukin-4 or interleukin-10. The TCL produced a wide range of interferon-gamma levels (138 to 15,020 pg/ml). Clones from 3 patients showed low levels (60 to 1,410 pg/ml), from 2 patients high levels (6,105 to 43,040 pg/ml) and from 1 patient a wide range (405 to 36,010 pg/ml) of interferon-gamma production. Thus epidermal CD8+ Tc1 cells in chronic plaque psoriasis produce highly heterogeneous levels of interferon-gamma, which may reflect clinical diversity.     

寻常型银屑病患者外周血髓系树突状细胞CD47的表达

目的：检测寻常型银屑病患者外周血髓系树突状细胞（mDCs）表面整合素相关蛋白（CD47）的表达水平。方法：流式细胞仪检测寻常型银屑病患者和健康对照者外周血mDCs表面CD47及CD4+调节性T细胞（Treg）表达水平；Luminex液相芯片技术检测上清液IL-23、IL-17和TNF-α浓度。结果：共检测46例患者和46名健康对照，患者组mDCs细胞CD47表达水平为32.8低于对照组的63.6，差异有统计学意义（P<0.05）。患者组mDCs细胞CD47表达水平与银屑病皮损面积和严重程度指数（PASI）负相关（r=-0.651, P<0.05）；患者组Treg细胞比例显著低于对照组，差异具有统计学意义（P<0.05）；IL-23、IL-17和TNF-α浓度显著高于对照组，差异具有统计学意义（P<0.05）。结论：银屑病外周血mDCs表面CD47及Treg细胞表达下降。     

IL-10 secretion of allergen-specific skin-derived T cells correlates positively with that of the Th2 cytokines IL-4 and IL-5*

Abstract In atopic individuals, allergen-specific CD4+ T lymphocytes often belong to the T-helper 2 (Th2) subset as they secrete the marker cylokincs interleukin-4 (IL-4) and IL-5 but not intcrfcron-y (INF-y). IL-10 is a cytokine the production of which, in the mouse system has been described to be restricted to the Th2 subset, but in the human was found to be produced by both ThI and Th2 T cell clones (TCC). We have recently shown that house dust mite antigen (Dermatophagoides pteron-yssinus)-specific TCC isolated from skin of patients with atopic dermatitis have a more polarized Th2 cytokine production profile than TCC obtained from the peripheral blood of these patients. In this study, we report that skin-derived TCC secrete more IL-10, IL-4 and IL-5, than TCC isolated from the blood of the same individual (p < 0.05). The difference was more significant with specific TCC than with non-specific TCC. Furthermore, there was a positive correlation between the production of IL-10 and that of IL-4 and IL-5, respectively. In addition, the amount of IL-4 and IL-5 secreted by specific TCC from the skin correlated positively. These results were confirmed by the detection of mRNA by PCR. Finally, our data confirm that in human blood-derived TCC IL-10 secretion is not related to a particular cytokine production profile. We suggest that the skin of AD provides an unique environment for the development of aTh2-likc secretion pattern not only with respect to IL-4 and IL-5 but also regarding IL-10.     

寻常性银屑病患者白介素20基因多态性研究

目的:选取位于1q32区域的白介素(IL)-20基因,通过检测基因序列研究基因多态性与银屑病的相关性.方法:提取银屑病患者(203例)和正常人(91名)基因组DNA并进行扩增,通过自动测序的方法测定IL-20基因及启动子区域的序列,检测其单核苷酸多态性(SNP)位点,并以SPSS软件进行统计学处理.结果:①IL-20启动子SNP位点-1723 C>G多态性的G等位基因频率,在以上呼吸道感染为发病诱因或加重因素的寻常性银屑病患者组和正常对照组间的频率分别为45.8%和32.4%,两组间差异有统计学意义(X2=5.539,P=0.019).结论:位于IL-20基因启动子区的SNP位点-1723C>G可能与以上呼吸道感染为发病诱因或加重因素的寻常性银屑病相关.     

寻常型银屑病患者miR-124与Th17 Treg细胞及相关细胞因子表达的关系

 目的探讨寻常型银屑病患者miR-124与Th17 Treg细胞及相关细胞因子表达的相关性。方法选取寻常型银屑病患者100例作为实验组（进行期47例,稳定期53例）,同时选取50例体检健康者作为对照组,采集外周血样本;用PCR法检测各组miR-124表达水平;用流式细胞仪检测Th17和Treg细胞数量;用双抗体夹心ELISA法检测各组细胞因子TNF-α、IFN-γ、TGF-β、IL-6、IL-17、IL-21、IL-23水平;采用RT-PCR法检测各组ROR-γt mRNA和Foxp3 mRNA转录水平,同时分析miR-124表达与Th17 Treg细胞及相关细胞因子表达的相关性。结果银屑病进行期组和稳定期组miR-124表达水平均低于对照组（均P<0.05）。对照组Th17、Treg和Th17/Treg水平分别为（0.64±0.31）%、（5.11±1.45）%和（0.13±0.05）,均低于银屑病进行期组和稳定期组患者（均P<0.05）。寻常型银屑病进行期和稳定期患者TGF-β的浓度分别为（476.52±34.42）、（471.10±52.39） pg/L,均低于对照组（均P<0.05）;而其他细胞因子TNF-α、IFN-γ、IL-6、IL-17、IL-21和IL-23水平均高于对照组（均P<0.05）。对照组患者ROR-γt mRNA和Foxp3 mRNA值分别为9.14±2.98、2.45±0.97,均高于银屑病进行期组和稳定期组患者（均P<0.05）。对照组miR-124水平与Th17和Treg之间无相关性（r=0.30,P=0.542;r=0.35,P=0.612）,而银屑病进行期组和稳定期组患者miR-124水平与Th17和Treg之间均存在相关性（P<0.05）。IFN-γ、TGF-β、IL-17、IL-21和IL-23与miR-124表达之间均存在相关性（均P<0.05）。结论寻常型银屑病患者Th17和Treg细胞的分化失衡与miR-124表达有关,miR-124表达异常减少可能是寻常型银屑病患者发生发展的因素之一。     

Foxp3+ regulatory T cells and related cytokines differentially expressed in plaque vs. guttate psoriasis vulgaris

Background Differences in the number of Foxp3+ regulatory T cells (Tregs) in lesional skin and peripheral blood and their functioning in plaque vs. guttate psoriasis have not been reported. Objectives To investigate whether there is a differential expression of Foxp3+ Tregs and a differential regulation of inflammatory cytokines in plaque vs. guttate psoriasis vulgaris. Methods The number and the percentage of Foxp3+ cells in different phases of skin lesions of patients with plaque and guttate psoriasis vulgaris were assessed by immunohistochemical staining. The expression of Foxp3 and interleukin (IL)‐17 protein in CD4 populations was measured by flow cytometry. Inflammatory cytokine production by transforming growth factor‐β1‐induced Foxp3+ Tregs was assessed in an in vitro study. The cytokines in supernatant and serum were determined by enzyme‐linked immunosorbent assay. Results The percentage of Foxp3+ CD3+ cells in the papillary layer was higher than in the reticular layer of dermis and in epidermis (P < 0·05). The numbers of Foxp3+ Tregs in skin lesions and peripheral blood were higher in plaque than in guttate psoriasis, whereas the percentage of IL‐17+ CD4+ cells was higher in guttate than in plaque psoriasis (P < 0·05). The numbers of Foxp3+ cells were positively correlated with the Psoriasis Severity Index score of skin lesions (P < 0·0001), and the percentages of Foxp3+ CD4+ cells in peripheral blood were positively correlated with the Psoriasis Area and Severity Index score of patients (P < 0·05). The inhibitory functions of Tregs to IL‐17 and IL‐6 in guttate psoriasis and to tumour necrosis factor‐α in plaque psoriasis were deficient. Conclusions Differential expression and regulatory functioning for inflammatory cytokine production by Foxp3+ Tregs may imply a different immunopathogenesis for plaque and guttate psoriasis.     

Cytokine gene polymorphisms in Chinese patients with psoriasis

BACKGROUND: Previous studies have shown that cytokine gene polymorphisms may confer susceptibility to psoriasis. OBJECTIVES: To determine whether genetic polymorphisms of the cytokine genes might influence the development of psoriasis in Chinese patients in Taiwan. METHODS: DNA samples were obtained from 170 patients with psoriasis vulgaris (PV), 102 patients with psoriatic arthritis (PsA) and 210 control subjects. Using direct sequencing and microsatellite genotyping, we examined 28 polymorphisms in 11 cytokine genes including the interleukin (IL)-1alpha, IL-1beta, IL-1 receptor antagonist, IL-4, IL-8, IL-10, IL-12B, IL-13, tumour necrosis factor (TNF)-alpha, TNF-beta and interferon-gamma genes. Genotypes of HLA-Cw*0602, killer cell immunoglobulin-like receptor (KIR) genes and major histocompatibility complex class I chain-related gene A (MICA) were also determined in patients with PsA. RESULTS: The patients with PV were more likely to carry the +4496G allele of the IL-12B gene (59.4% vs. 49.3%, P = 0.0067, P(c) = 0.033). However, no significantly different allelic and genotypic distributions of the other analysed genes including IL-1beta, TNF-alpha, TNF-beta, KIR genes and MICA were found between the PV/PsA patients and controls. Moreover, no association was observed with disease onset, gender, peripheral arthritis or joint erosion. With regards to HLA-Cw*0602, its allele frequency was significantly increased in patients with early-onset PV (25.3% vs. 4.8%, P < 10(-7)), but not in patients with PsA. CONCLUSIONS: The IL-12B gene polymorphism conferred a risk for PV in our Chinese population, although the effect was more minor than that of HLA-Cw*0602. Cw*0602, KIR2DS1/S2 and MICA-A9 were unlikely to be risk alleles in our patients with PsA. The other analysed genetic polymorphisms of cytokine genes do not appear to be associated with susceptibility to PV and PsA in Chinese patients in Taiwan.     

Positive treatment effects of ustekinumab in psoriasis: Analysis of lesional and systemic parameters

Ustekinumab, a human anti‐interleukin (IL)‐12/IL‐23p40 monoclonal antibody has demonstrated significant efficacy in patients with moderate‐to‐severe psoriasis. Skin lesion biopsies, cell surface markers on peripheral blood lymphocytes, and ex vivo T‐helper (Th)1/Th2 cytokine responses from peripheral blood mononuclear cells (PBMC) from patients receiving ustekinumab 45 or 90 mg, or placebo were evaluated at baseline and week 12. Inflammatory serum protein levels were measured at baseline, week 2 and week 12. At week 12, median epidermal thickness decreased from 312.1 to 132.7 μm, and median levels of cellular proliferation (Ki67) and T‐cell infiltration (CD3) decreased by 84.3% and 70.7%, respectively, in the combined ustekinumab group (all P ≤ 0.002). Serum levels of tumor necrosis factor (TNF)‐α, C‐C motif ligand 27 (CCL27) and other inflammatory cytokines remained unchanged. Minimal variation in the percentage of T cells expressing cutaneous lymphocyte antigen (CLA) was observed following ustekinumab treatment, with no significant variation in the percentage of cells expressing CD45RA, CD45RO, CD25, human leukocyte antigen‐DR (HLA‐DR), and C‐X‐C motif receptor 3 (CXCR3). No apparent effect on the magnitude of Th1/Th2 responses to external stimuli in PBMC was observed following placebo or ustekinumab treatment. Ustekinumab improves histological psoriasis measures, with minimal impact on the systemic immune system.     

Local effects of TL01 phototherapy in psoriasis

The mechanisms whereby narrowband ultraviolet B (UVB) (311-313 nm, TL01) phototherapy are effective in psoriasis may differ from those occurring in broadband UVB phototherapy. In the present study, changes in epidermal cells as a result of TL01 therapy were assessed in the skin of patients with psoriasis. The non-lesional skin of five subjects with plaque psoriasis was biopsied before and after a series of 12 whole-body TL01 treatments. Following appropriate staining of skin sections, the numbers of p53-positive keratinocytes, sunburn cells and Langerhans cells in the epidermis were counted. TL01 therapy induced a threefold increase in the number of p53-positive epidermal cells, a 12-fold increase in sunburn cells and a twofold decrease in Langerhans cells. The increase in epidermal p53 expression and apoptosis of keratinocytes together with the depletion of Langerhans cells in the non-lesional skin of psoriasis patients are likely to contribute to the effectiveness of TL01 phototherapy.     

Immunologic implications of PUVA therapy in psoriasis vulgaris

Summary During the combined effects of psoralen and UVA irradiation (PUVA therapy) a significant decrease (P<0.005 in T cells has been noted in 10 psoriasis patients and 10 healthy controls especially after four exposures. Based on the fact that the total number of circulating lymphocytes of the patients and the PUVA-treated healthy controls remained within the normal range, this decrease may be due to temporary physicochemical changes of the membranes of these cells but not to T cell lysis. After eight exposures these decreased T cell values returned to starting range. The starting T cell range in psoriasis patients is significantly lower (P<0.005) compared to that of healthy controls.It is of importance that before PUVA therapy in all the patients antibodies reactive with a basal cell nuclear antigen belonging to the four main Ig classes (IgM, IgD, IgE, and IgA) could be removed from the membrane of circulating lymphocytes by means of acid elution. In contrast, mainly the IgA antinuclear basal cell antibody could be eluted from circulating PMN leukocytes in the patients under investigation. After eight PUVA exposures, however, corresponding antibodies, belonging to the three main Ig classes (IgM, IgD and IgE) could also be eluted from the membranes of circulating PMN-leukocytes of the same patients. This implies an exchange of molecules during photochemotherapy. Finally, it could be shown that after effective PUVA therapy antinuclear basal cell antibodies of the psoriasis patients under study were reactive not only with the nuclei of the basal cell layer but also with almost all the nucleic of the epidermis of the uninvolved and lesional skin. The latter finding points to the fact that PUVA treatment causes at least antigenic changes of nuclear proteins in all the nuclei of the epidermis of the PUVA treated skin. Moreover, inflammatory cells with Fab fragments within the cells present in the lesional skin before PUVA disappear during this treatment.Based on a poster session presented at the occasion of the seventh joint meeting of the ESDR, Amsterdam, The Netherlands, May 1977     

寻常性银屑病患者血清白细胞介素-26水平与疾病严重度相关性分析

目的:检测寻常性银屑病患者血清中白细胞介素(IL)-26的表达量,并分析其对疾病严重程度的影响.方法:选取2017年12月-2019年1月来山西医科大学第一附属医院就诊的寻常性银屑病患者84例作为观察组,其中进展期患者40例,静止期患者44例;选择同期在该院进行健康体检的志愿者40例作为对照组.采用流式细胞术检测2组患...     

CD4~+CD25~+调节性T细胞与Th17细胞在寻常型银屑病中的表达

目的:检测外周血中CD4~+CD25~+调节性T细胞(简称Treg细胞)与Th17细胞在寻常型银屑病(PV)中的表达。方法:34例寻常型银屑病患者,18例正常人作为对照。采用流式细胞术检测外周血中Treg细胞、FOXP3~+Treg细胞及Th17细胞的表达水平。结果:PV患者外周血中Treg细胞比例和FOXP3~+Treg细胞比例分别为(3.68±1.22)%和(0.53±0.19)%,低于正常对照组的(6.63±1.00)%和(0.76±0.14)%(P0.05)。Th17细胞比例为(2.20±0.78)%,高于正常对照组的(0.65±0.22)%(P0.05)。PV患者外周血中Treg细胞的表达与FOXP3~+Treg细胞的表达呈正相关(r=0.563,P0.05),而Treg细胞的表达与Th17细胞的表达呈负相关(r=-0.522,P0.05);Treg细胞和Th17细胞二者与PASI评分无相关性,与病程亦无相关性。结论:Treg细胞表达的减少及Th17细胞应答的增强,可能是导致PV免疫失衡的重要原因。     

Dramatic improvement of psoriatic erythroderma after acute hepatitis: analysis of cytokine synthesis capability in peripheral blood T cells

We report a patient with psoriasis and hepatitis C virus infection who initially presented with psoriatic erythroderma and eventually showed complete clearance of psoriatic lesions following acute hepatitis induced by etretinate treatment. Cytokine synthesis capabilities in peripheral blood T cells obtained at different stages were evaluated in this patient. A dramatic increase in the frequency of interferon-gamma-producing CD8+ T cells in peripheral blood was observed during the erythrodermic stage. In contrast, the frequencies of interleukin (IL)-4- and IL-13-producing CD4+ T and CD8+ T cells were remarkably high at the resolution stage. These results clearly indicate that a shift towards type 2 cytokine predominance contributes to the resolution of severe psoriasis.     

寻常型银屑病患者外周血中微粒子的表达

目的：检测微粒子在寻常型银屑病外周血中的表达。方法：高速离心法分离33例寻常型银屑病患者和33名正常人对照外周血T细胞、B细胞及单核细胞来源的微粒子,分别进行细胞特异性单克隆抗体标记,采用流式细胞仪检测三种细胞来源的微粒子数量。结果：银屑病组患者血浆中T淋巴细胞（CD3+）来源的微粒子、单核细胞（CD14+）来源的微粒子和B淋巴细胞（CD19+）来源的微粒子数量分别为（6.6±0.2）×105,（3.9±0.3）×105和（4.1±0.2）×105高于正常对照组的（2.0±0.3）×105,（1.6±0.3）×105和（1.1±0.2）×105.治疗后银屑病组患者微粒子的总数量较治疗前明显减少（P<0.01）。此外,银屑病患者血浆中微粒子水平与PASI评分具有相关性,尤其是T淋巴细胞源性微粒子。结论：微粒子的数量在寻常型银屑病外周血中明显升高,可能在银屑病的发病中起到重要作用。     

清热凉血汤对寻常性银屑病患者Th1／Th2型细胞因子的影响

目的观察清热凉血汤对寻常性银屑病患者Th1／Th2型细胞因子的影响。方法将88例患者随机分为两组，对照组口服阿维A胶囊，治疗组在此基础上给予清热凉血方。结果疗效观察：治疗8周后．两组疗效比较差异有统计学意义（P〈0．05），治疗8周后干扰素（IFN）-γ、白介素（IL）-2两组均有明显下降（P〈0．05），但治疗组下降更为明显（P〈0．05），与对照组相比有统计学意义（P〈0．05），治疗组IL-4、IL-10有明显升高（P〈0．05），与对照组相比有统计学意义（P〈0．05）。结论清热凉血汤对于Th1／Th2免疫反应的影响，主要是抑制Th1型免疫反应．提高Th2型免疫反应从而使银屑病患者失衡的Th1/Th2型反应相对地向Th2型免疫反应方向改变而达到临床症状的缓解。     

寻常型银屑病患者体内半乳糖凝集素家族蛋白表达研究

 目的:探讨寻常型银屑病（PV）患者体内半乳糖凝集素（Gal）家族蛋白表达情况。方法:采用ELISA法检测30例PV患者与30例正常对照者血清中Gal-1、Gal-3、Gal-7、Gal-9的水平;采用免疫组化检测15例PV患者皮损与10例正常皮肤中Gal-1、Gal-3、Gal-7、Gal-9的表达及细胞定位;以平均光密度值评价表达水平,比较其在PV患者与正常对照组中表达的差异。结果:与正常对照者相比,PV患者血清中Gal-1水平明显上调［（39.57±11.33） ng/mL比（31.93±7.98） ng/mL,t=3.02,P=0.004）］,且与PASI评分呈正相关（r=0.37,P=0.049）;皮损中Gal-1主要表达于真皮且表达明显上调（0.030±0.043比0.011±0.002,t=3.34,P=0.003）;PV患者表皮中Gal-3表达下调（0.016±0.001比0.060±0.018,t=-2.85,P=0.009）、 Gal-7上调（0.090±0.011比0.049±0.009,t=2.58,P=0.017）、 Gal-9下调（0.011±0.002比0.049±0.007,t=-6.16,P<0.001）。结论:Gal家族成员Gal-1、Gal-3、Gal-7、Gal-9在PV患者血清中的水平及皮损局部的细胞定位与表达各不相同,提示其在PV中的功能也存在差异,有可能参与了PV的发病机理。     

51例新疆寻常型银屑病患者皮损中FOXP1和hMSH2的表达

目的:检测新疆寻常型银屑病(PV)患者皮损中FOXP1和hMSH2的表达。方法:EnVision免疫组化法分别检测51例新疆PV患者皮损及29例正常人皮肤组织中FOXP1和hMSH2的表达。结果:FOXP1和hMSH2在PV皮损中的阳性表达率(92.16%,94.12%)均高于正常人皮肤组织(65.52%,55.17%),差异有明显统计学意义(P0.01)。结论:FOXP1和hMSH2可能与寻常型银屑病的发病有关。