染料木黄酮抑制小鼠变应性接触性皮炎的实验研究

目的： 探讨染料木黄酮(Gen)对小鼠变应性接触性皮炎(ACD)的抑制作用。 方法： 建立二硝基氟苯(DNFB)诱发的小鼠ACD模型，观测不同剂量Gen对小鼠耳廓肿胀程度、鼠耳皮肤组织病理变化、体重及胸腺指数、脾指数的影响。 结果： 各剂量Gen组均能明显抑制DNFB诱发的小鼠耳廓肿胀，抑制炎症细胞浸润，降低胸腺指数，且不影响小鼠体重增长。低剂量Gen组增加脾指数，高剂量Gen组降低脾指数。 结论： Gen对DNFB诱发的小鼠ACD有明显抑制作用。     

氧化苦参碱抑制二硝基氟苯所致小鼠接触性皮炎及淋巴细胞增殖

目的：探讨氧化苦参碱(OMT)对二硝基氟苯(DNFB)所致小鼠变应性接触性皮炎(ACD)的抑制作用及小鼠淋巴细胞增殖的影响。 方法： ①建立DNFB所致小鼠ACD模型，以不同剂量的OMT、PBS、氢化可的松(HCT)进行腹腔注射，检测小鼠耳廓肿胀度变化。②利用羧基荧光素乙酰乙酸(CFDA-SE)染色，流式细胞术检测OMT对小鼠淋巴细胞增殖的影响。 结果： ①OMT对DNFB所致小鼠ACD呈剂量依赖性抑制作用,且与同等剂量HCT作用效果相似,但副作用明显减小。②体外实验证明，在500、125和31 mg/L组，OMT对小鼠淋巴细胞增殖呈剂量依赖性抑制。 结论: OMT对DNFB所致小鼠ACD有显著的抑制作用，而且抑制小鼠淋巴细胞增殖；OMT是一种免疫抑制剂。     

喘可治注射液对人外周血单个核细胞Th1/Th2细胞因子谱的影响

目的：通过研究喘可治注射液对人外周血单个核细胞（PBMCs）Th1／Th2细胞因子谱的影响，探讨喘可治注射液的免疫调节作用机制。方法：以流式微球分析（CBA）法检测不同处理情况下，人外周血单个核细胞分泌Th1（IFN-γTNF-α、IL-2）和Th2（IL4、IL-6、IL-10）细胞因子水平。结果：健康人PBMCs体外培养12小时后，上清中细胞因子主要为TNF-α和IL-6，喘可治使Th1和Th2细胞因子全面升高；喘可治对PDB加离子霉素诱导的PBMCs分泌Th1和Th2细胞因子具有抑制作用，并能抑制流感病人异常升高的INF-γ、TNF-α、IL-6和IL-10分泌。结论：喘可治注射液上调健康人PBMCs分泌TH1和Th2细胞因子，对异常活化的PBMCs分泌的Th1和Th2细胞因子则具有下调作用。     

甘草甜素抑制小鼠接触性超敏反应的实验研究

目的：探讨甘草甜素(GL)对二硝基氟苯(DNFB)所致小鼠接触性超敏反应(CHS)的抑制效果。 方法： BALB/c小鼠分为GL1(11 mg/kg)组、GL2(22 mg/kg)组、GL3(44 mg/kg)组、生理盐水(NS)组和地塞米松(DXM)组5组；利用DNFB诱发小鼠右耳CHS，各组于 -1 d 至5 d每天腹腔注射0.2 mL上述不同剂量药物(溶于生理盐水)。以耳厚度差、耳重量差及右耳病理变化，观察GL对小鼠CHS的抑制效果；同时计算胸腺指数、脾指数，了解GL对CHS小鼠免疫系统的影响。 结果： 3个剂量组GL和DXM均可明显抑制CHS小鼠右耳厚度及重量的增加，与NS组间差异显著(P<0.01)，同时亦可明显抑制右耳组织水肿和炎细胞浸润。3种剂量GL均可减低CHS小鼠胸腺重量，与NS组间有统计学差异(P<0.05)，而对脾脏重量无明显影响；DXM可显著降低CHS小鼠胸腺和脾脏重量。 结论： 甘草甜素可明显抑制DNFB诱发的小鼠接触性超敏反应。     

喘可治延长小鼠异基因移植皮片存活时间与CD4+CD25+调节性T细胞相关性研究

目的：研究喘可治抑制小鼠异基因皮片移植排斥作用与小鼠体内CD4+CD25+ 调节性T细胞（CD4+CD25+Tr）变化的相关性。 方法： 建立小鼠同种异基因与同基因皮片移植模型，通过腹腔注射给药喘可治(CKZ)观察其对皮片移植存活时间的影响，并利用3色免疫荧光标记和流式细胞术分析受鼠外周血CD4+CD25+ Tr变化规律。 结果： 同种异基因移植CKZ组的移植皮片存活时间显著长于同种异基因移植对照组，分别为（195±23）d和（102±22）d，P＜001；在同种异基因皮片移植后，受体外周血CD4+CD25+ Tr占CD4+T细胞百分率明显升高，术后8 d达到高峰（P＜001），然后出现下降趋势，其中同种异基因移植对照组在术后13 d时已回落至正常水平，而同种异基因移植CKZ组在术后23 d时仍维持在高于移植前水平；在同基因皮片移植后，CKZ组与对照组外周血CD4+CD25+ Tr水平均无明显升高（P＞005）。 结论： 喘可治可延长小鼠同种异基因移植皮片存活时间，通过升高CD4+CD25+ Tr水平而发挥免疫抑制效应可能是其机制之一。     

变应性接触性皮炎的免疫调节、评估和干预新进展

变应性接触性皮炎(ACD)的发生发展取决于刺激反应与抑制反应的平衡,此过程存在众多的调节因素,包括血管紧张素转换酶、神经免疫皮肤系统、应激、肥大细胞等因素,并由此衍生各种相应的免疫评估和免疫干预,免疫评估主要有体内测定法和体外测定法等,免疫干预则主要通过细胞因子介导。     

迟发型超敏反应中小鼠脾淋巴细胞与细胞外基质粘附能力的变化

目的:探讨迟发型超敏反应中淋巴细胞与细胞外基质粘附能力的变化及细胞因子的调节作用。方法: 以2, 4, 6-三硝基氯苯(picrylchloride, PCl)两次致敏小鼠后, 在耳部攻击造成超敏反应。取攻击后不同时间点的脾淋巴细胞, 以Mn²⁺为诱发剂, 检测细胞与细胞外基质蛋白的粘附活性变化;或取攻击后18h的脾淋巴细胞, 分别在体外与IL-2, IFN-γ, TNF-α单独或联合培养4h后, 检测细胞与细胞外基质的粘附作用;将脾淋巴细胞纯化为T细胞后, 检测TNF-α对其粘附能力的影响。结果:脾淋巴细胞与细胞外基质的粘附能力在攻击后6h时开始上升, 18h达到高峰, 之后下降, 在36h基本恢复正常水平。IL-2在10×10⁴U/L显著促进脾淋巴细胞的粘附, TNF-α剂量依赖性地促进脾淋巴细胞的粘附, IFN-γ对TNF-α的作用具有明显的协同效应。TNF-α对脾T细胞粘附能力的促进作用强于对总脾淋巴细胞的作用。结论:迟发型超敏反应中, T淋巴细胞与细胞外基质的粘附随炎症的进展发生相应的变化, 这种粘附作用受各种细胞因子的调节。     

治喘贴对实验豚鼠嗜酸粒细胞凋亡和对白细胞介素-5的影响

目的　研究治喘贴对卵清蛋白所致实验豚鼠哮喘体内嗜酸粒细胞 (EOS)凋亡及对白细胞介素 (IL 5 )的调节作用。方法　将实验动物随机分为正常对照组、哮喘模型组、代温灸膏治疗组、治喘贴治疗组。经外贴治疗 18d ,收集支气管肺泡灌洗液 (BALF)并采集心脏血 ,采用流式细胞仪、免疫组织化学 (TUNEL)和酶联免疫吸附 (ELISA)法分析血中EOS凋亡和BALF中EOS凋亡数及IL 5含量。结果　哮喘豚鼠血和BALF中EOS凋亡数、IL 5含量均高于正常对照组动物 (P均 <0 .0 1) ,治喘贴治疗豚鼠IL 5和EOS均明显低于哮喘模型组 (P均 <0 .0 1)。结论　治喘贴能促进哮喘豚鼠体内EOS凋亡 ,并降低IL 5的水平 ,从而改善哮喘模型豚鼠的气道变态性炎症反应。     

艾叶挥发油治疗大鼠变应性鼻炎的实验研究

目的:观察艾叶挥发油对变应性鼻炎(AR)大鼠血清中IgE、IL-4和IL-5含量的影响。方法:60只SD大鼠按体重随机分为正常对照组、模型组、治疗组和阳性药对照组,每组10只。实验组用卵蛋白(OVA)、氢氧化铝和灭活百日咳杆菌皮下注射致敏,用10%的卵蛋白生理盐水溶液滴鼻激发,建立AR模型。正常对照组以等量生理盐水代替。治疗组和阳性药对照组分别灌胃给予艾叶挥发油和氯雷他定治疗10天,正常对照组用橄榄油代替药物;另外20只大鼠用于被动皮肤过敏原试验(PCA),检验大鼠模型制备及治疗情况;用ELISA法测定各组大鼠外周血清IL-4、IL-5和IgE含量。结果:模型组大鼠鼻喷嚏平均次数明显多于正常对照组、治疗组和阳性对照组(P<0.05);模型组大鼠PCA试验阳性率为92%,其余三组全部表现为阴性;模型组IgE均显著高于正常对照组(P<0.05);治疗组IgE、IL-4、IL-5均显著低于模型组(P<0.05);治疗组各项指标与阳性对照组比较差异无统计学意义。结论:艾叶挥发油可降低AR大鼠血清中IL-4、IL-5和IgE含量,减轻鼻粘膜变应性炎症。     

国产虫草素(cordycepin)抗小鼠迟发型超敏反应的实验研究

目的：研究国产虫草素抗小鼠迟发型超敏反应的作用及其免疫机理。方法：用2，4-二硝基氯苯（2，4-DNCB）对昆明种小鼠皮肤致敏和诱发，制成迟发型超敏反应模型。以国产虫草素溶液作为实验组，分为两个剂量组（实验组1：12mg／kg；实验组2：16mg／kg），以生理盐水作为阴性对照组，所有溶液均为间隔24小时（最后一次间隔为4小时）重复腹腔注射给药。计算激发后各组小鼠左右耳耳廓肿胀厚度差、耳廓增重值及肿胀度，进行统计分析。并经过HE染色观察虫草素对小鼠致炎耳炎症病理变化及脾脏病理变化的影响。结果：两种给药剂量的国产虫草素溶液对模型鼠红斑，水肿。渗出的接触性皮炎损害有明显抑制作用，可减轻由于充血、水肿等炎症反应导致的局部皮损增厚及重量的增加。与生理盐水对照组相比。两实验组虫草素溶液的抗炎作用均有显著性差异（厚度差：P〈0．0001；肿胀度：P〈0．05）；两实验组间的抗炎作用也有显著性差异（厚度差及肿胀度均为P〈0．05），且与给药剂量相关。三组小鼠脾指数未见明显差异（P〉0．05），脾脏组织病理变化未见明显差异。结论：虫草素以24小时间隔（最后一次给药间隔为4小时）经腹腔注射后，可能通过其免疫调节作用对迟发型超敏反应引起的小鼠接触性皮炎发挥明显的抑制效应，该效应与给药剂量相关，同时对脾脏组织未见明显毒性作用。     

A case of autoimmune progesterone dermatitis misdiagnosed as allergic contact dermatitis

Autoimmune progesterone dermatitis is a rare autoimmune response to endogenous progesterone that usually occurs in fertile females. Cutaneous or mucosal lesions develop cyclically during the luteal phase of the menstrual cycle when progesterone levels are elevated. Symptoms usually start 3-10 days before menstruation and resolve 1-2 days after menstruation ceases. We report the case of a 48-year-old woman with intermittent eczematous skin lesions of the legs, forearms, and buttocks. She was diagnosed with allergic contact dermatitis, and topical steroids were prescribed. Her skin eruptions waxed and waned for 6 years and were associated with her menstrual cycle. We performed an intradermal test using progesterone, which was positive, and prescribed gonadotropin-releasing hormone analogues monthly for 3 months. The patient's skin lesions improved, confirming the diagnosis. Autoimmune progesterone dermatitis should be included in the differential diagnosis of recurrent eczema that is refractory to treatment in women of child-bearing age.     

Dipeptidyl peptidase IV (DPP4) deficiency increases Th1‐driven allergic contact dermatitis

Background CD26 or dipeptidyl peptidase IV (DPP4) is known to be involved in several immunological processes and has recently been reported to play a crucial role in the allergic responses of the lungs. Objectives To explore the impact of DPP4 on the allergic response of the skin. Methods Skin biopsies from patients suffering from atopic dermatitis (AD) and healthy controls were investigated for the expression of CD26/DPP4. Furthermore, the functional impact of CD26 was investigated in two models of contact hypersensitivity using CD26/DPP4‐deficient and wild‐type rats. Dinitrochlorobenzene (DNCB) was used to induce a T helper type 1 (Th1)‐dominated inflammation and toluene‐2,3‐diisocyanate for a Th2‐pronounced inflammation. The inflammatory responses were determined by histological quantification, flow cytometry [fluorescence‐activated cell sorting (FACS)], and an enzyme‐linked immunosorbant assay (ELISA). Results CD26/DPP4‐expression was up‐regulated in the lesional skin biopsies of patients compared with healthy controls as well as in both models of contact hypersensitivity. However, in the more Th2‐driven model, a reduced inflammatory skin response was found in CD26/DPP4‐deficient rats, analogous to the effects observed recently in a rat model of asthma. In partial contrast, there was an aggravation of local skin inflammation in CD26/DPP4‐deficient rats under conditions of Th1‐like skin inflammation. Conclusion and Clinical Relevance The up‐regulation of CD26 in atopic dermatitis represents a new finding, which has also been seen in other inflammatory skin diseases. However, tissue expression of CD26/DPP4 in immunological skin response can either be beneficial or aggravating, depending on a possible Th1/Th2 shift. This might have consequences for humans suffering from diabetes mellitus treated by DPP4 inhibitors, who have eczematous skin diseases as a co‐morbidity. Cite this as: T. Tasic, W. Bäumer, A. Schmiedl, F. Schwichtenhövel, R. Pabst, U. Raap, S. von Hörsten and M. Stephan, Clinical & Experimental Allergy, 2011 (41) 1098–1107.     

Mechanisms of chemical-induced innate immunity in allergic contact dermatitis

Allergic contact dermatitis (ACD) is one of the most prevalent occupational skin diseases and causes severe and long-lasting health problems in the case of chronification. It is initiated by an innate inflammatory immune response to skin contact with low molecular weight chemicals that results in the priming of chemical-specific, skin-homing CD8(+) Tc1/Tc17 and CD4(+) Th1/Th17 cells. Following this sensitization step, T lymphocytes infiltrate the inflamed skin upon challenge with the same chemical. The T cells then exert cytotoxic function and secrete inflammatory mediators to produce an eczematous skin reaction. The recent characterization of the mechanisms underlying the innate inflammatory response has revealed that contact allergens activate innate effector mechanisms and signalling pathways that are also involved in anti-infectious immunity. This emerging analogy implies infection as a potential trigger or amplifier of the sensitization to contact allergens. Moreover, new mechanistic insights into the induction of ACD identify potential targets for preventive and therapeutic intervention. We summarize here the latest findings in this area of research.     

Contribution of CD4+ and CD8+ T-cells in contact hypersensitivity and allergic contact dermatitis

Allergic contact dermatitis, also referred to as contact hypersensitivity, is one the most frequent inflammatory skin diseases, and is characterized by redness, papule and vesicles, followed by scaling and dryness. Allergic contact dermatitis is elicited upon skin contact with nonprotein chemicals, haptens, and corresponds to a cutaneous delayed type hypersensitivity reaction, mediated by hapten-specific T-cells. During the sensitization phase, both CD4⁺ and CD8⁺ T-cell precursors are activated in the draining lymph nodes by presentation of haptenated peptides by skin dendritic cells. Subsequent hapten painting on a remote skin site induces the recruitment and activation of specific T-cells at the site of challenge. This leads to apoptosis of keratinocytes, recruitment of inflammatory cells and development of clinical symptoms. Experimental studies from the last 10 years have demonstrated that, in normal contact hypersensitivity responses to strong haptens, CD8⁺ type 1 T-cells are effector cells of contact hypersensitivity through cytotoxicity and interferon-γ production, while CD4⁺ T-cells are endowed with downregulatory functions. The latter may correspond to the recently described CD4⁺CD25⁺ regulatory T-cell population. However, in some instances, especially when there is a deficient CD8⁺ T-cell pool, CD4⁺ T-cells can be effector cells of contact hypersensitivity. Ongoing studies will have to confirm that the pathophysiology of human allergic contact dermatitis is similar to the mouse contact hypersensitivity and that the contact hypersensitivity response to common weak haptens, most frequently involved in human allergic contact dermatitis, is similar to that reported for strong haptens.     

Results of standard series patch testing in patients with occupational allergic contact dermatitis

BACKGROUND: In the workplace, the skin is at high risk of exposure to chemicals and other contaminants, and occupational dermatitis is an important field of study. METHODS: We evaluated 230 patients referred to our clinic because they were affected by dermatitis suspected to be of occupational and allergic origin. They were tested with the allergens of the GIRDCA standard series, and with supplementary series when indicated. RESULTS: Among the 230 subjects, 49 were positive only to supplementary series (26.3% of all allergic contact dermatitis), while 130 (69.9% of all allergic contact dermatitis) were considered to have an occupational allergic contact dermatitis diagnosed by the standard series alone. The most frequent occupations of the patients were health care workers and hairdressers/beauticians. The most common agents responsible for occupational allergic contact dermatitis were metals and para-phenylenediamine. CONCLUSIONS: As the standard series detected a relatively low proportion of occupational allergic contact dermatitis, it is not adequate to recognize an occupational allergic contact dermatitis, certain supplementary series should also to be tested. However, even if such occupational series are used, we remain likely to continue to underestimate the frequency of occupational allergic contact dermatitis, because workers come into contact with a large number of substances that are often unknown.