尖锐湿疣患者Th1/Th2细胞因子的检测

目的 检测尖锐湿疣(CA)患者CD3⁺T细胞分泌细胞因子的水平,分析Th1细胞因子和Th2细胞免疫活动。方法 首先用刺激物刺激细胞,增加细胞内细胞因子的表达,然后加入荧光标记的特异性抗细胞因子单克隆抗体,特异性抗原抗体结合,最后应用流式细胞仪分析特异性细胞因子表达水平。结果 CA患者Th1型细胞因子干扰素γ、白介素2、肿瘤坏死因子α、白介素12表达水平较正常对照显著降低,Th2型细胞因子白介素10表达水平较正常人对照差异无显著性。结论 CA患者Th1型反应模式处于弱势状态,相对地Th2型反应模式处于优势状态,Th1/Th2平衡失调,Th1/Th2平衡向Th2方向漂移。CA患者体内Th2型细胞因子模式占优势状态,可能是病毒发生免疫逃逸引起免疫损伤的一种机制。     

基于Logistic回归分析尖锐湿疣患者复发的影响因素及与皮损局部辅助性T细胞1/辅助性T细胞2免疫失衡的相关性

目的 探讨尖锐湿疣(CA)患者复发的影响因素及与皮损局部辅助性T细胞1/辅助性T细胞2(Th1/Th2)免疫失衡的相关性。方法 选取2019年1月至2020年6月第四军医大学西京皮肤医院收治的224例CA患者作为研究对象。随访1年,统计复发率,并根据复发情况分为复发组和未复发组。比较两组的基线资料、Th1细胞因子[白介素-2(IL-2)、干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)]和Th2细胞因子[白介素-4(IL-4)、白介素-6(IL-6)、白介素-10(IL-10)]水平。构建预测复发的Logistic回归模型,分析模型及Th1/Th2细胞因子的预测价值。结果 文化程度、IL-2、IFN-γ、TNF-α是复发的保护因素,饮酒、性伴侣数量、性伴侣伴有CA、经常熬夜、IL-4、IL-6、IL-10是复发的危险因素(P<0.05);Logistic回归模型预测复发的曲线下面积(AUC)为0.742;IL-2、IFN-γ、TNF-α、IL-4、IL-6、IL-10联合预测复发的AUC为0.890。结论 文化程度、饮酒、性伴侣数量、性伴侣伴有CA、经常熬夜及皮损局部Th...     

尖锐湿疣患者外周血CD4~+ T细胞细胞因子的检测

目的检测尖锐湿疣(CA)患者外周血CD4+T细胞细胞因子IL-2,IL-12,IFN-γ和IL-4的水平,探讨其在CA发病中的作用。方法应用流式细胞仪对60例CA患者和20例正常对照者外周血CD4+T细胞细胞因子IL-2,IL-12,IFN-γ,IL-4的表达进行了检测。结果CA患者外周血IL-2,IL-12,IFN-γ-CD4+T细胞百分率均显著下降(P<0.001);IL-4-CD4T细胞百分率与正常对照组比较差异无显著性(P>0.05);Th1/Th2比值显著低于正常对照组(P<0.001)。结论CA患者存在Th1/Th2平衡失调,这可能是CA发生、发展的重要原因之一。     

寻常型银屑病皮损中Th17细胞相关因子的表达

目的:研究Th17细胞相关因子在银屑病发病机制中的作用.方法:用逆转录-聚合酶链反应法(RT-PCR)半定量分析寻常型银屑病患者皮损区、非皮损区及正常人皮肤组织中Th17细胞相关因子IL-17和IL-23(p19/p40)、IL-6 mRNA的表达.结果:银屑病患者皮损中IL-17、IL-23p19、IL-23p40和IL-6的 mRNA平均相对表达量分别为1.231±0.843、1.166±0.142、1.125±0.104和1.186±0.222.皮损组织中该4种细胞因子mRNA表达水平均高于非皮损组和正常皮肤组,非皮损组四者表达高于正常对照组,差异均有统计学意义(均P<0.05).结论:Th17细胞相关因子在银屑病患者皮损中过度表达,提示Th17型细胞因子在银屑病的免疫发病机制中可能起着非常重要的作用.     

系统性红斑狼疮患者Th1/Th2细胞及白介素12、18及其受体mRNA的表达

目的 探讨未经药物治疗的初发系统性红斑狼疮患者Th1/Th2细胞亚群分布,白介素12、白介素18及其受体的基因表达.方法 运用三色荧光标记流式细胞术检测35例初发SLE患者、10例正常人Th1/Th2细胞亚群分布;ABI7700Real-TimePCR法同时检测38例患者和28例正常人IL-12、IL-18及其受体mRNA表达水平.结果 ①初发狼疮患者Th1较正常人明显减低(P<0.05),但Th1/Th2无显着性改变.②与正常人组相比,SLE组患者IL-12、IL-18mRNA及其受体表达较正常人明显降低(P值均<0.05);③面部红斑组患者Th1/Th2、IL-12P35较正常人组降低(P均<0.05);④RNP阳性组患者IL-12P40较正常人组升高,IL-12P35、IL-18较正常人组降低(P均<0.05).结论 SLE是一种以Th1细胞下降,Th2细胞相对占优势的自身免疫性疾病,源于诱导向Th1细胞分化的一系列细胞因子及其受体减少和细胞因子间失衡所致.     

尖锐湿疣患者MyD88表达水平与血清TNF-α、IL-10的关系

目的:分析尖锐湿疣患者MyD88表达水平与血清TNF-α、IL-10的关系,探索尖锐湿疣的发病机制。方法:选择30例尖锐湿疣(CA)患者作为研究对象,15例正常包皮环切者作为对照组。使用免疫组织化学法检测CA患者皮损组织、对照组包皮组织中MyD88的表达水平,使用ELISA法检测所有研究对象血清TNF-α和IL-10水平。结果:免疫组化分析发现76%(23/30)的CA患者的皮损组织中呈现MyD88阳性表达,积分为5.01±0.48。CA患者两种血清炎症因子的水平均显著高于正常对照组[TNF-α:(2.09±1.35)μg/L vs(1.03±0.32)μg/L,P0.05;IL-10:(29.05±15.88)μg/L vs(18.38±5.99)μg/L,P0.05]。MyD88表达水平与TNF-α、IL-10呈正相关(r_(TNF-α)=0.23,P0.05;r_(IL-10)=0.46,P0.05)。结论:CA患者皮损组织的MyD88表达水平以及血清TNF-α、IL-10水平均显著升高,MyD88表达与TNF-α、IL-10呈正相关,提示CA患者免疫抑制与免疫损伤同时存在。     

光动力疗法对尖锐湿疣皮损朗格汉斯细胞的影响

目的观察光动力疗法(PDT)对尖锐湿疣(CA)患者皮损中朗格汉斯细胞(LC)的影响,进一步探寻PDT治疗CA的免疫机制。方法应用免疫组化标记方法,对15例PDT治疗前后的CA皮损及5例正常包皮组织对照进行染色,在高倍光镜下观察LC的形态、分布及数量等的变化。结果 15例CA皮损较正常对照组织[(18.8±4.0)个/高倍视野]LC数量明显降低(P0.01),表皮内LC分布不规则,大部分LC结构不完整,细胞突减少、缩短甚至消失,缺少LC的典型特征,点状或条索状。个别皮损表皮内LC缺失,而其真皮内见到不典型的LC。PDT治疗后即刻组LC数量[(9.2±2.0)个/高倍视野]较治疗前组[(6.2±1.8)个/高倍视野]明显升高(P0.05),治疗后7 d组LC数量[(6.4±1.1)个/高倍视野]回落接近治疗前水平(P0.05),差异无统计学意义。结论 CA患者皮损局部细胞免疫功能低下,作为抗原提呈细胞的LC,其数量和形态发生改变。PDT能诱导CA皮损局部LC数量及形态发生明显改变,具有调节局部免疫的作用。     

Th17细胞相关因子与寻常性进行期银屑病的相关性研究

目的探讨Th17细胞相关因子白细胞介素(IL)-17A、IL-17F、IL-21、IL-22与寻常性进行期银屑病发病的相关性。方法通过实时定量反转录聚合酶链式反应(RT-PCR)分别检测30例患者和20名正常人外周血单个核细胞(PBMC)、12例患者皮损、12名正常皮肤组织中上述4种细胞因子的mRNA表达水平。结果患者组PBMC中IL-17A、IL-17F、IL-21和IL-22的mRNA表达水平较正常组显著升高(P均0.05),患者组皮损中4种细胞因子的mRNA表达明显高于正常组(P均0.05)。结论 Th17细胞因子IL-17A、IL-17F、IL-21和IL-22的mRNA水平在患者组PBMC及皮肤组织中明显升高,提示Th17细胞因子可能与寻常性银屑病的发病有一定相关性。     

阿维A对寻常性银屑病皮损组织中Th1/Th2平衡的影响

目的:通过对阿维A治疗前后寻常性银屑病(PV)患者皮损中Th1、Th2细胞的研究,了解局部Th1、Th2、Th1/Th2平衡在PV病情发展中的变化.探讨阿维A治疗PV的作用机制.方法:采用免疫荧光双重标记法检测健康对照、阿维A治疗前后PV患者皮损中Th1(CD3+IFN-γ+)、Th2(CD3+IL-4+)细胞的数量变化.结果:①阿维A治疗前后的PV患者皮损真皮浅层T细胞分别为(24.03±8.03)个和(15.32±5.26)个、Th1细胞分别为(15.57±6.73)个和(7.66±4.28)个,数量较健康对照组增多IT细胞(5.44±0.17)、Th1细胞(3.78±0.98)个],P<0.05;Th2细胞数量在治疗前后分别为(6.29±8.39)个和(7.46±10.28)个,与健康对照组(2.44±0.77)个相比,差异无统计学意义,P>0.05;②Th1/Th2平衡在治疗前明显偏向于Th1一方,治疗前比值为6.49±8.40,治疗后为1.86±1.75,与健康对照1.58±0.25相比差异无统计学意义;③Th1/Th2平衡比例与银屑病皮损面积和严重度指数(PASI)评分呈正相关,治疗前相关系数r=0.86,治疗后相关系数r=0.69.结论:PV患者皮损处T细胞遵循1h1型免疫应答理论:Th1/Th2平衡与PASI相关;阿维A治疗PV有效,可使PASI降低;阿维A治疗PV时可能通过干预Th1/Th2平衡而起作用.     

尖锐湿疣局部细胞免疫功能状态与人乳头瘤病毒型的相关性研究

我们采用 (1)聚合酶链反应检测皮损中HPV6 11、16 18。 (2 )应用免疫组化技术对 6 1例CA患者疣体组织、疣旁“正常”组织及 2 0例对照包皮组织表皮中CD1a+ 朗格汉斯细胞 (LC)及表真皮中CD4 +T、CD8+ T细胞数目及CD4 + CD8+ 比值进行检测。探讨CA患者局部细胞免疫功能状态与HPV型的关系。结果CA疣体组织表皮CD1a+ LC细胞数量较正常对照明显降低 (P <0 .0 1) ,不同HPV亚型感染所致CA其CD1a+ LC数目差异无显著性 (P >0 .0 5 ) ;真皮中CD4 + T、CD8+ T细胞数目较正常对照升高 ,但CD4 + CD8+ 比值下降 (P <0 .0 1) ;CD4 + T、CD8+ T细胞数目及CD4 + CD8+ 比值相应的改变与不同HPV亚型无关。表皮CD1a+ LC与表皮、真皮中CD4 + T细胞及CD4 + CD8+ 比值成正相关 ,与CD8+ T细胞数目成负相关。因此 (1)CA患者皮损局部细胞免疫功能低下 ;疣体组织中CD1a+ LC细胞数目及CD4 + CD8+比值均降低。 (2 )CA患者局部免疫缺陷程度与HPV型无相关性。     

Skin scratching switches immune responses from Th2 to Th1 type in epicutaneously immunized mice

BACKGROUND: The balance between Th1 and Th2 subsets is important with respects to susceptibility and resistance to particular infection or autoimmune diseases. However, the mechanism controlling Th1/Th2 balance remains unclear, although several factors have been reported to induce Th1/Th2 differentiation. Atopic Dermatitis (AD) that is a chronic skin disorder has been known as Th2 biased nature characterized by high expression of IgE in the serum. In contrast, the chronic skin lesions express IFNgamma and some patients don't show IgE in the serum. Thus, the pathology is also complicated now. OBJECTIVE: We focused on skin scratching that is common feature in the patients. In this study, we investigated in order to determine whether skin scratching regulates immune responses in murine epicutaneous sensitization model. METHODS: The scratched mice on abdominal skin using wire brush were applied with keyhole limpet hemocyanin (KLH) on the skin using occlusive patch. We examined the immune responses including delayed type hypersensitivity (DTH) reaction, antigen-specific serum immunoglobulin formation, and cytokine expressions on the local skin in comparison with mice without scratching. RESULTS: We found that the epicutaneously sensitized mice with KLH on abdominal skin showed Th2 biased immune response including expression of antigen-specific IgE in the serum and IL-13 in the local skin. Surprisingly, scratching on local abdominal skin using wire brush exchanged the immune response from Th2 dominance to Th1, because the mice displayed DTH reaction and significant level of antigen-specific IgG2a and IgG2b but not IgE in the serum. Furthermore, the abdominal skin showed significant level of IFNgamma but not IL-13. CONCLUSION: These data demonstrate that skin scratching switches immune response from Th2 biased response to Th1. This suggests that skin scratching play critical roles as one of exogenous immune modulator. This murine sensitization model may help to understand natures of several allergic disorders including AD.     

Lipoteichoic acid-related molecule derived from the streptococcal preparation, OK-432, which suppresses atopic dermatitis-like lesions in NC/Nga mice

Bacterial stimulation may serve to control atopic disorders such as atopic dermatitis (AD) through inducement of Th1 cell-mediated immune response. The lipoteichoic acid (LTA)-related molecule (okLTA) from streptococcal preparation, OK-432, has been shown to be a potent Th1 inducer through the action of IL-12. Examination was made of the therapeutic effects of this okLTA injected intra- and/or subcutaneously into AD-like lesions in NC/Nga mice, particularly in the vicinity of the suppressor of cytokine signaling (SOCS) regulatory pathways. Using immunohistochemical staining with IL-4/IL-12p40 and phosphorylated STAT6/p-STAT4 and RT-PCR for IL-4/IL-12p40, STAT6/STAT4 and mRNA expression and in situ hybridization of SOCS3 and 5, evaluation was made of the immunoregulatory effects of this okLTA in the treatment of spontaneous AD-like lesions in NC/Nga mice. Following the injection of okLTA, remarkable improvement in the lesions of NC/Nga mice was noted. In okLTA-treated skin, IL-12p40/p-STAT4 positive cellular infiltration was extensive while IL-4/p-STAT6 positive cell infiltration was seen to diminish considerably, compared to untreated NC mice. SOCS3 in situ expression in okLTA-treated mice was noted to be significantly less compared to untreated NC mice, in which the expression was prominent. SOCS5 in situ expression was rather, though not significantly, strong in okLTA-treated mice. okLTA treatment is clearly shown to induce Th1 cellular response and down-regulate immune response in the Th2 pathway through SOCS3 reduction in AD-like lesions of NC/Nga mice. The present results demonstrate that bacterial wall components such as okLTA should serve as an effective new therapeutic approach for treating AD.     

复发与未复发女性尖锐湿疣患者Th_1/Th_2细胞因子检测

目的观察复发与未复发女性生殖器尖锐湿疣(CA)患者外周血Th1/Th2细胞因子水平,探讨CA复发与Th1/Th2细胞因子的相关性。方法选取女性生殖器CA初发患者37例,经电灼治疗后随访3个月(期间不使用任何免疫制剂),再次复发者计入A组(15例),未复发者计入B组(22例)。用ELISA法测定电灼治疗前各组血清标本的细胞因子IL-2、IL-12、IL-4和IL-10的浓度,比较A,B两组血清细胞因子及Th1/Th2比值的差异,分析CA复发与Th1/Th2细胞因子水平的相关性。结果A组治疗前的血清IL-2,IL-12,IL-4,IL-10分别为65.86±20.31,21.09±5.84,98.31±10.95,25.87±9.24,B组分别为82.81±27.24,25.74±10.63,89.02±13.80,25.62±9.84;用Th1/Th2比值来预测CA不复发的敏感性为86.4%,特异性为80.0%,ROC曲线下面积为0.812。结论CA复发与否与患者血清Th1/Th2细胞因子水平呈负相关,以Th1/Th2比值预测CA复发与否有较高的敏感性和特异性,Th/Th比值可作为评价CA复发与否的一个参考指标。     

Study on the T-helper cell 1/2 cytokine profile in blister fluid of patients with herpes zoster and its clinical significance

Herpes zoster (HZ) is a Varicella zoster virus infection disease. Previous studies have presumed the connection between development of HZ and involvement of cellular immunity in peripheral blood. However, whether cellular immunity plays a role in the local skin lesion has not been addressed. To explore the levels of T-helper cell (Th)1/Th2 type cytokine profiles in the blister fluid of the skin lesions from the patients with HZ and its role in pathogenesis, we used the cytometric bead array kit to compare the levels of cytokines (interleukin [IL]-2, tumor necrosis factor [TNF]-α, IL-10 and IL-4) in blister fluid from 46 patients with those from the suction blister fluids from 20 volunteers without any infectious disease (the control group). The results indicated that the levels of Th1 cytokines, IL-2 and TNF-α in the blister fluid from the patients' skin lesions were significantly lower than those from the control group, whereas the levels of Th2 cytokines IL-10 and IL-4 were significantly higher than those in the control group. Moreover, significant variation of the levels of Th1/Th2 cytokines (IL-2, TNF-α, IL-10 and IL-4) in the blister fluid from the HZ patients' lesions was also observed among different stages of the disease. It is concluded that a cytokine imbalance was present in the local lesions of patients with HZ during disease development. Our data suggested that the Th immunity was associated with disease activity, which may play an important role in the pathogenesis of HZ.     

Therapeutic effects of streptococcal preparation OK-432 on atopic dermatitis-like lesions in NC/Nga mice: possible shift from a Th2- to Th1-predominance

BACKGROUND: The inducement of Th1 cell-mediated immune response, possibly brought about through bacterial stimulation, may serve to control atopic disorders such as atopic dermatitis (AD). The streptococcal preparation, OK-432, has been shown a potent Th1 inducer through the action of IL-12. NC/Nga mice under ordinary conditions have been found to contract dermatitis similar to human AD. OBJECTIVE: Examination was made of the therapeutic effects of OK-432 local intra- and/or subcutaneous injections on AD-like lesions in NC/Nga mice. METHODS: Immunohistochemical staining with IL-4/IL-12p40 and CD80/86 and phosphorylated STAT4/p-STAT6 and RT-PCR for IL-4/IL-12p40 and STAT6/STAT4 mRNA was conducted for the evaluation of OK-432 treatment of spontaneous AD-like lesions in NC/Nga mice. RESULTS: At 5 weeks following injection of OK-432, for treating head and back lesions in NC/Nga mice, 10 of 12 OK-432 treated NC mice were found to have clinically improved quite considerably. On the head and back skin of OK-432-treated mice, IL-12p40/CD80 positive cellular infiltration was conspicuous, in contrast to non-treated mice. IL-4/CD86 positive cellular infiltrates in OK-432-treated mice had decreased significantly more than in non-treated mice and IL-4 mRNA expression was virtually absent in OK-432-treated mice. P-STAT4 positive cells could be seen abundantly present in OK-432-treated mice, and p-STAT6 positive cells were much fewer than in non-treated mice. CONCLUSIONS: OK-432-treatment appears to induce Th1 cellular response and to down-regulate that of the Th2 pathway in AD-like lesions of NC/Nga mice. The present results demonstrate bacterial components from such Streptococcus to likely constitute an effective new therapeutic approach in the treatment of AD.     

复发性尖锐湿疣细胞因子mRNA检测结果分析

目的：检测复发性尖锐湿疣患者(CA)疣体内多种细胞因子mRNA的表达，研究Th1／Th2型细胞因子在CA复发中的相互关系。方法：采用多探针核糖核酸酶切保护法检测12例复发性CA疣体IL-2，IFN-gamma，IL-4，IL-5，IL-9，IL-10，IL-13，IL-15mRNA的表达。结果：从被保护探针的放射性强弱发现复发性CA患者IL-2，／FN-gamma，IL-15的mRNA无表达，而IL-4，IL-5，IL-9，IL-10，IL-13的mRdNA表达。结论：从基因水平进一步证实复发性CA患者疣体内存在Th1／Th2细胞因子平衡失调，Th2型细胞因子表达基因普遍增强，机体免疫反应由Th1型细胞因子强势转为Th2型细胞因子强势可能在CA复发的发病机理中起着重要作用。     

Expression of cytokines and chemokine receptors in the cutaneous lesions of erythema multiforme and Stevens-Johnson syndrome/toxic epidermal necrolysis

BACKGROUND: Erythema multiforme (EM) and Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN) are caused by a dysregulation of cellular immunity. OBJECTIVES: To evaluate further the potential role of certain cytokines and chemokine receptors in cutaneous lesions of patients affected by EM and SJS/TEN and to establish whether such diseases are polarized preferentially towards a T-helper (Th) 1 or Th2 pattern. METHODS: Biopsy specimens from eight patients with EM, six with SJS/TEN and three healthy controls were stained for immunohistochemical examination using the alkaline phosphatase-antialkaline phosphatase method. The monoclonal antibodies used included those to tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, interleukin (IL)-2, IL-5, IL-13, receptor 3 for C-C chemokines (CCR3), receptor 3 for C-x-C chemokines (CXCR3) and CXCR4. RESULTS: The SJS/TEN specimens showed significantly higher expression of all the cytokines and chemokine receptors (except CXCR3) tested than the EM specimens. Both lesional series showed significantly higher expression of all the receptors tested than the healthy control specimens, with the sole exception of a lower expression of CCR3 in EM specimens. Comparison between molecules associated with a Th1 or Th2 response revealed a predominance of Th1 response in EM and no significant imbalance between Th1 and Th2 in SJS/TEN. CONCLUSIONS: We have provided further evidence that TNF-alpha is strongly expressed in SJS/TEN lesions and therefore it may be involved in the epidermal necrosis featured in such diseases. IFN-gamma may play an important role both in EM and SJS/TEN. IL-2, IL-5 and IL-13 may contribute to the cutaneous immunoinflammation in these diseases. Chemokine receptors may be involved strongly in the recruitment of inflammatory cells in lesional skin. In our cases we found a sharp polarization towards a Th1 pattern in EM, while the SJS/TEN lesions showed a mixed Th1/Th2 pattern.     

Expression of IL-18 in psoriasis

Abstract Interleukin-18 (IL-18) is a novel cytokine that plays an important role in the T-helper 1 (Th1) response, primarily via its ability to induce IFN-γ production in T cells and NK cells. Human keratinocytes produce IL-18, as do monocytes and macrophages, which are the two major sources of this molecule. It is thought that IL-18 derived from keratinocytes might be involved in the cutaneous Th1-type immune response. In the present study, we investigated the expression of IL-18 in psoriatic lesional skin and attempted to determine whether immunoreactive IL-18 in crude extracts of psoriatic scales is processed to the mature, active form. Immunohistochemical and RT-PCR analysis showed that the expression of IL-18 was increased in psoriatic lesional skin relative to that in normal skin. Western blotting and an ELISA for IL-18 in combination demonstrated that the immunoreactive IL-18 in extracts of psoriatic scales contained the mature form of IL-18, but most of the IL-18 was pro-IL-18. No bioactivity of IL-18 or IFN-γ inducibility in human PBMC could be detected in psoriatic scales. Taken together, these findings indicate that keratinocyte-derived IL-18 participates in the development of the Th1 response in psoriatic lesions, and that its bioactivity appears to be tightly regulated in cutaneous inflammation. Received: 18 October 2000 / Revised: 3 February 2001 / Accepted: 27 April 2001