雷公藤多甙对变应性接触性皮炎患者外周血调节性T细胞的影响

目的：通过检测变应性接触性皮炎（ACD）患者经雷公藤多甙（TG）治疗前后外周血CD4^＋、CD25^＋、CD127^-调节性T细胞（Treg）占总T淋巴细胞的百分比的变化,初步探讨TG治疗该病的作用机制。方法：选取斑贴试验确证镍为主要过敏原的ACD患者,随机纳入TG试验组52例和外用润肤霜对照组30例,在治疗前和治疗2周后,分别采患者外周血,通过流式细胞仪检测患者的Treg的百分比,同时观察患者的临床疗效。结果：TG治疗组患者外周血Treg占总T细胞的百分比较治疗前明显升高（P〈0．05）,并且患者症状有效改善;对照组治疗前后各指标比较无统计学差异。结论：TG提高ACD患者外周血Treg的百分比,可能是其发挥治疗作用的机制之一。     

调节性T细胞在变应性接触性皮炎中的作用

调节性T细胞是一类具有免疫调节作用的T细胞亚群。近年来研究发现调节性T细胞除了CD4+CD25+Foxp3+T细胞外,还有多种亚型,通过多种免疫调节机制作用于效应细胞和抗原提呈细胞发挥免疫抑制作用,参与机体多种炎症和自身免疫相关疾病。变应性接触性皮炎是一种常见的炎症性皮肤病,是抗原特异性T细胞介导的皮肤超敏反应。调节性T细胞通过多种免疫调节通路,在其致敏阶段和诱发阶段均发挥重要作用,从而阻止弱抗原引发的接触性皮炎,减弱强抗原引发的皮肤炎症反应。     

干扰素诱导蛋白10对变应性接触性皮炎小鼠T细胞亚群的影响

目的 探讨干扰素诱导蛋白10(IP-10)对体外培养变应性接触性皮炎(ACD)小鼠T淋巴细胞亚群的影响,探讨ACD的发病机制.方法 建立ACD小鼠模型,分离纯化小鼠脾淋巴细胞,并和不同浓度的IP-10共同培养,应用流式细胞术检测与IP-10共同孵育的小鼠脾淋巴细胞中CD4+、CD8+和CD3+水平,以方差分析进行结果　分析.结果 IP-10各浓度组体外培养的ACD小鼠CD4+、CD8+、CD3+T淋巴细胞的含量均高于对照组,但仅高浓度组与对照组之间的差异具有统计学意义(P<0.01).结论 IP-10可升高成熟T淋巴细胞比率,非特异性地增加Ⅳ型变态反应主要效应细胞CD4+T及CD8+T亚群的表达,扩大变应性接触性皮炎的免疫反应.     

特应性皮炎患儿外周血维生素D水平与辅助性T细胞/调节性T细胞相关性分析

目的:探讨特应性皮炎(AD)患儿血清中25羟维生素D_3[25(OH)D_3]水平与AD严重程度、外周血中辅助性T细胞(Th17)及调节性T细胞(Treg)表达水平的相关性分析。方法:选取2018年4月—2019年6月来该院皮肤科就诊的71例AD患儿(AD组),采用配对的方式选取该院儿科71例健康体检儿童作为对照组,利用特应性皮炎积分指数评分(SCROAD)评估AD组患儿疾病严重程度,并检测2组血清25(OH)D_3、外周血Th17及Treg水平。结果:AD组患儿25(OH)D_3含量及Treg表达水平均显著低于对照组;而Thl7表达水平明显高于对照组,差异均有统计学意义(P0.05)。AD组患儿血清25(OH)D_3水平与SCORAD评分呈负相关(r=-0.41,P0.05),外周血Treg水平与血清25(OH)D_3含量呈正相关(r=0.37,P0.05),外周血Th17水平与血清25(OH)D_3含量呈负相关(r=-0.41,P0.05)。结论:维生素D(VitD)水平与AD疾病严重程度相关,VitD水平可能与Th17/Treg失衡相关。     

接触性皮炎的发病机制研究进展

接触性皮炎的发病涉及到表皮角质形成细胞、效应T细胞以及炎症因子的相互作用.除了特异反应性T细胞,不依赖抗原提递细胞提呈的淋巴细胞亚群,也在皮肤变态反应的启动和快速放大中起重要作用.本文就近年来角质形成细胞、免疫细胞及炎症因子在接触性皮炎发病机制中的研究进展做一综述.     

特应性皮炎患者外周血CD4+CD25+调节性T细胞的检测

目的 探讨CD4+CD25+调节性T细胞（CD4+CD25+ Treg）在特应性皮炎（AD）发病中的作用机制及临床意义。方法 流式细胞仪分析AD患者外周血中CD4+CD25+ Treg细胞数量,实时荧光定量PCR检测外周血单核细胞（PBMC）中Foxp3 mRNA水平,ELISA检测血清中IL-2、IL-4、IL-10、IFN-γ水平。结果 AD患者外周血中CD4+CD25+ Treg细胞占CD3+ T细胞及CD4+ T细胞的比例均明显低于正常人对照组（t′ = 3.775、4.533,P值均 ＜ 0.01）;外周血中CD4+CD25+ Treg细胞占CD3+ T细胞比例在AD患者急性期明显低于慢性期（t = 2.217,P < 0.05）,而在急性期与亚急性期、亚急性期与慢性期之间差异均无统计学意义（t = 1.558、0.49,P值均 ＞ 0.05）。AD患者PBMC中Foxp3 mRNA的水平低于正常人对照组（z = -2.368,P ＜ 0.05）;其外周血中CD4+CD25+ Treg细胞与血清中IL-2和IL-10成正相关（r = 0.512、0.494,P值均 ＜ 0.05）,与IL-4和IFN-γ的相关性无统计学意义（r = -0.110、-0.237,P值均 ＞ 0.05）。结论 在AD患者中,外周血中CD4+CD25+ Treg细胞数量及Foxp3 mRNA水平均下降,从而可能减少对Th2细胞增生及其细胞因子分泌的抑制,使Th2占优势,参与AD的发病。     

外用茶油对变应性接触性皮炎发生的影响

目的 观察茶油对小鼠变应性接触性皮炎发生的影响。方法 建立二硝基氟苯致小鼠变应性接触性皮炎模型,外用药物进行治疗,观察发生皮炎的小鼠耳片的组织学变化（HE染色）,RT-PCR检测组织中炎症细胞因子IL-2、IFN-γ mRNA的表达水平。结果 成功建立二硝基氟苯致小鼠变应性接触性皮炎的动物模型,且皮肤中IL-2、IFN-γ mRNA的表达显著增高。哈西奈德溶液组和茶油组的耳片组织肿胀明显减轻,炎症反应轻微,炎性细胞浸润数目少,组织病理变化轻。在茶油和哈西奈德溶液的干预下,炎性组织IL-2、IFN-γ的mRNA表达均不同程度下降。结论 茶油减轻二硝基氟苯致小鼠变应性接触性皮炎的病理损害,下调了炎症因子的表达。     

变应性接触性皮炎模型鼠体内Th17细胞活性变化

目的:研究变应性接触性皮炎(allergic contact dermatitis,ACD)模型鼠外周血及皮损处Th17细胞活性变化情况及其意义。方法:利用2,4-二硝氟苯(DNFB)诱导构建ACD小鼠模型,分别于激发前及激发后的第3、6、9、12、24 h取小鼠耳组织及外周血,评估小鼠耳组织炎症情况,采用实时荧光定量PCR法检测小鼠皮损处IL-17及RORγt mRNA的表达水平,利用酶联免疫吸附试验检测血清中IL-17水平。结果:激发后24 h内小鼠耳组织肿胀程度逐渐加重,皮损中IL-17和RORγt mRNA表达量也逐渐升高,激发后各组与激发前组比较差异有统计学意义(均P<0.05);耳组织IL-17和RORγt mRNA表达量与其肿胀程度呈正相关;激发后小鼠血清中IL-17水平升高,在6小时达到最高,随后逐渐下降。结论:在ACD小鼠的急性炎症期,皮损及外周血中Th17细胞活性增强,并且局部皮损处的Th17细胞活性与炎症程度密切正相关。     

过敏性接触性皮炎反应中浸润T细胞类型研究

为了观察过敏性接触性皮炎（ＡＣＤ）反应浸润Ｔ细胞的类型，本实验用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）及免疫组化方法对１８例ＡＣＤ急性期患者皮损进行了白介素２ｍＲＮＡ及白介素２、γ干扰素、白介素４表达的研究。结果：与对照组织相比，绝大多数ＡＣＤ标本有Ｔｈ１型细胞因子的表达，且定位于真、表皮浸润的淋巴细胞胞浆中。结果提示ＡＣＤ反应是以Ｔｈ１型细胞增殖为主的反应。     

用曲泰斯贴剂检测变应性接触性皮炎

目的 观察曲泰斯斑贴试剂在我国应用的可行性。方法 对100例皮肤病门诊患者用曲泰斯贴剂进行斑贴试验研究。其中确诊变应性接触性皮炎患者21例，非变应性接触性皮炎患者19例，可疑变应性接触性皮炎60例。结果 确诊变应性接触性皮炎组获与临床符合的阳性结果19例；非变应性接触性皮炎组阳性4例，均与临床无关。斑贴试验的灵敏度为90.5%，特异度78.9%，准确性85.0%，除轻度瘙痒外，未见不良反应。结论 曲泰斯贴剂在我国检测变应性接触性皮炎安全有效。     

变应性接触性皮炎患者外周血淋巴细胞趋化因子受体CCR5表达的研究

目的：探讨趋化因子受体5（CCR5）在接触过敏发病中的作用。方法：采用免疫荧光标记一流式细胞仪检测了25例变应性接触性皮肤炎患者外周血淋巴细胞在培养前，后CCR5的表达，结果：接触性皮炎患者的淋巴细胞在培养前，后CD^4,CCR^5细胞均分别高于正常对照组，有显著差异（P＜0．05和P＜0．01）；淋巴细胞培养48小时后，接触性皮炎患者的CD^4,CCR^5高于培养前（P＜0．01），而正常对照组则低于培养前（P＜0．01），结论：接触性皮炎患者的T细胞表达Th1淋巴细胞的表面标志CCR5的能力增强，有利于Th1细胞归巢到Th1型炎症反应部位。     

Increased “in vivo” lymphocyte blastogenesis in the peripheral blood of patients with atopic dermatitis and allergic contact dermatitis

Summary The spontaneous ³H-thymidine (³HT) labelling of some lymphocyte subpopulations has been studied in the peripheral blood of five patients with atopic dermatitis and five with widespread allergic contact dermatitis and compared with that in 10 healthy subjects. One hour after addition of ³HT to heparinized blood, lymphocytes were separated and processed with two different rosetting techniques (E-rosette test and Active-E-rosette test). The cell suspensions were cytocentrifuged and autoradiography undertaken.An increased number of ³HT labelled lymphocytes was observed in the peripheral blood of patients with dermatitis as compared to controls. These labelled lymphocytes were E-rosette-forming cells (T cells) and E-non-rosetteforming cells (non-rosette-forming T cells and non-T cells). The ratio between the labelling index (LI) of E-rosette- to the LI of non-E-rosette-forming cells was in favour of T cells in allergic contact dermatitis (ratio=3.09) whereas in atopic dermatitis (ratio=0.93) the DNA synthesis was relatively greater in the non-rosette-forming cells.It is suggested that this increased LI of peripheral blood lymphocytes could be related to the increased dermal mononuclear cell ³HT-labelling that has been reported previously in these inflammatory skin diseases.D. Van Neste is recipient of a grant from INSERM, Paris and Administration des relations culturelles internationales (ARCI), Bruxelles     

尖锐湿疣患者外周血T细胞抗原受体γδ T细胞的检测

目的：探讨外周血T细胞抗原受体（TCR）γδT细胞在尖锐湿疣发病机制中的作用。方法：采用双色荧光抗体染色技术经流式细胞仪检测20例尖锐湿疣患者外周血TCRγδT细胞与TCRαβT细胞的百分率。结果：尖锐湿疣患者外周血中TCRγδT细胞百分率比正常对照组显著增高（P＜0．01），TCRαβT细胞的百分率与正常对照组相比差异无显著性（P＞0．05）。结论：尖锐湿疣患者外周血TCRγδT细胞数量增多是机体在受到病毒感染时的一种早期非特异性免疫反应，在尖锐湿疣的发病机制中起一定的免疫防卫作用。     

IgE-positive epidermal Langerhans cells in allergic contact dermatitis lesions provoked in patients with atopic dermatitis

Summary To see whether or not IgE-bearing epidermal Langerhans cells are specific to skin lesions of atopic dermatitis (AD), we performed immunohistochemical and immunoelectron microscopic examinations of dinitrochlorobenzene (DNCB) contact dermatitis lesions provoked in uninvolved skin of eight patients with AD. In all of the eight examined, IgE-positive epidermal Langerhans cells were observed in the DNCB dermatitis lesions. Typical staining of anti-IgE was absent in the epidermis of normal-appearing skin of five patients with AD. Thus, it is likely that IgE positive epidermal Langerhans cells non-specifically occur in different eczematous diseases provoked in patients with AD.     

慢性荨麻疹患者外周血CD4+CD25+调节性T细胞的表达

目的:探索调节性T细胞(regulatory T cells,Tregs)在慢性荨麻疹(CU)发病中的作用.方法:采用流式细胞术分别检测CU患者和健康人外周血CD3+、CD4+、CD8+、CD4+CD25+T细胞的表达水平.结果:与健康对照组比较,CU患者外周血CD3+T细胞比例无明显变化,CD4+T细胞增高(P ＜ 0.05),CD4+CD25+T细胞表达水平显著增高(P ＜ 0.01),CD8+T细胞数量降低(P ＜ 0.05),CD4+/ CD8+比值明显升高(P ＜ 0.05).自身血清皮肤试验(ASST)阳性组与阴性组的CU患者CD4+CD25+Tregs 细胞的数量无统计学差异.结论:CU患者外周血存在Treg细胞及其他T淋巴细胞亚群的数量异常和分化失衡,这一异常在CU发病中的作用值得进一步研究.     

Impaired responses of peripheral blood mononuclear cells to nickel in patients with nickel-allergic contact dermatitis and concomitant atopic dermatitis

BACKGROUND: Allergic contact dermatitis (ACD) is pathogenetically dependent on cell-mediated immune responses mediated by type 1 T lymphocytes. Atopic dermatitis (AD), in contrast, occurs as a result of sustained activation of type 2 subsets of T cells. Although atopic patients may become sensitized to various contact allergens, little is known about the influence of atopy on delayed-type hypersensitivity. OBJECTIVES: To investigate the in vitro responses of peripheral blood mononuclear cells (PBMC) to nickel stimulation in groups of atopic and nonatopic patients with patch test-verified nickel ACD. METHODS: Ten nonatopic patients with nickel ACD, 10 patients with nickel ACD and concomitant AD, 10 patients with AD but with no contact allergy, and 10 healthy persons participated in the study. PBMC were cultured in the presence or absence of nickel sulphate, phytohaemagglutinin (PHA) or tetanus toxoid (TT). [(3)H]thymidine incorporation was used to measure the rate of antigen-induced DNA synthesis and enzyme-linked immunosorbent assay was used to measure the production of interleukin (IL)-2 (type 1 cytokine) and IL-5 (type 2 cytokine). RESULTS: Nickel-stimulated PBMC of nickel-allergic patients with AD proliferated significantly less and secreted significantly lower amounts of IL-2 than cells of nonatopic nickel-allergic patients. IL-5 production was also lower in the former group, although the difference was nonsignificant. Moreover, neither the nickel-specific DNA synthesis nor the cytokine production by PBMC of atopic nickel-allergic patients differed significantly from those of healthy control persons and AD patients without contact allergy. Proliferative and secretory responses of PBMC to PHA or TT stimulation differed nonsignificantly between the groups. Nickel-induced IL-2 production correlated well with IL-5 production in nickel-allergic patients regardless of their atopic status. CONCLUSIONS: Our results indicate that PBMC of nickel-allergic patients with concomitant AD are characterized by impaired in vitro proliferative and secretory responses to the contact allergen nickel but not to the mitogen PHA or the recall antigen TT. The type 2 cytokine IL-5 may play a role in the development of ACD.     

梅毒血清固定患者外周血调节性T细胞/Th17细胞平衡的研究

目的 探讨梅毒血清固定患者外周血调节性T细胞(Treg细胞)和Th17细胞平衡的变化.方法 梅毒血清固定患者26例,正常人对照23例,利用流式细胞仪分别检测外周血Treg细胞和Th17细胞的比例及相关特异性转录因子Foxp3和ROR-γt的定量表达情况,并进行相关性分析.结果 梅毒血清固定患者外周血CD4+T细胞中,Treg细胞比例为33.28％±11.84％,明显高于正常人对照组(22.13％±7.79％,P＜0.01);Th17细胞比例(3.17％±2.32％)显著低于正常人对照组(8.87％±2.00％,P＜ 0.01);CD4+T细胞内转录因子Foxp3的表达量(2994.86±1099.18)高于正常人对照组(2539.72±1086.96,P＜0.05),而ROR-γt的表达量(1473.12±752.20)低于正常人对照组(1778.34±388.13,P＜ 0.05).在所有受试者外周血CD4+T细胞中Foxp3和ROR-γt的表达呈负相关(r=-0.48,P＜0.01).结论 梅毒血清固定患者外周血存在Treg/Th 17细胞平衡的异常.