外周血T细胞亚群及细胞因子对外源性哮喘IgE生成…

对３０例哮喘患者及３０例健康成年人的外周血，采用单克隆抗体（ＭｃＡｂ）间接免疫荧光法测定Ｔ细胞亚群；ＥＬＩＳＡ双抗体夹心法测定ＩｇＥ、ＩＬ－４；ＦＩ２细胞株，生物学方法测定ＩＬ－２；ＩＬ－６依赖细胞株７ＴＤ１，掺入法测定ＩＬ－６；用抗人ＣＤ２３的ＭｃＡｂ测定ＣＤ２３。为研究Ｔ细胞、细胞因子对哮喘ＩｇＥ生成调节机理及细胞因子在哮喘发病过程中的作用。结果显示：发作期ＩｇＥ、ＩＬ－２、ＩＬ－４、ＣＤ２３     

肾病综合征患者PBMC上清IL－4和IFN－γ及血清IgE水平

本文报道，对４２例原发性单纯型肾病综合征（ＩＮＳ）患者及相应年龄组健康儿童进行ＩＬ－４、ＩＦＮ－γ及血清ＩｇＥ水平测定，结果表明，极期患者ＩＬ－４和ＩｇＥ水平均显著高于正常对照组，且两者呈正相关，而ＩＦＮ－γ水平显著低于正常对照组；缓解期患者ＩＬ－４、ＩＦＮ－γ及ＩｇＥ均接近于正常水平。提示ＩＬ－４及ＩＦＮ－γ为ＩｇＥ异常变化的两个重要细胞因子，对ＩｇＥ的合成，ＩＬ－４及ＩＦＮ－γ起着正负反馈的调节作用。     

外周血T细胞亚群及细胞因子对外源性哮喘IgE生成的作用

对３０例哮喘患者及３０例健康成年人的外周血，采用单克隆抗体（ＭｃＡｂ）间接免疫荧光法测定Ｔ细胞亚群；ＥＬＩＳＡ双抗体夹心法测定ＩｇＥ、ＩＬ－４；ＦＩ２细胞株，生物学方法测定ＩＬ－２；ＩＬ－６依赖细胞株７ＴＤ１，掺入法测定ＩＬ－６；用抗人ＣＤ２３的ＭｃＡｂ测定ＣＤ２３。为研究Ｔ细胞、细胞因子对哮喘ＩｇＥ生成调节机理及细胞因子在哮喘发病过程中的作用。结果显示：发作期ＩｇＥ、ＩＬ－２、ＩＬ－４、ＣＤ２３、ＣＤ８~＋、ＣＤ４／ＣＤ８~＋比值较对照组及缓解期有显著性差异（Ｐ＜０．０１）。缓解期ＩｇＥ与对照组之间无显著性差异（Ｐ＞０．０５）；ＣＤ８＋、ＣＤ４／ＣＤ８比值，ＣＤ２３与对照组之间有显著性差异（Ｐ＜０．０１）。ＣＤ２３、ＣＤ４、ＩＬ－６三组间无显著性差异（Ｐ＞０．０５）。结果表明，ＩｇＥ合成增加是哮喘发作的关键，Ｔ细胞对日细胞合成ＩｇＥ的调节是通过细胞因子实现的，细胞因子又参与气道炎症过程。     

FcεRⅡ／CD23在支气管哮喘发病中作用的研究

本文以急性发作期病人和缓解期病人及正常人各２０例为研究对象，探讨ＩＬ－４－ＦｃεＲⅡ／ＣＤ２３－ＩｇＥ在支气管哮喘中的调节作用。结果表明，急性发作期病人的ＩＬ－４分泌细胞、ＦｃεＲⅡ／ＣＤ２３阳性淋巴细胞和血清总ＩｇＥ水平均明显高于缓解期病人和正常对照组。在急性发作时，ＩＬ－４和ＣＤ２３之间存在着正相关。进一步表明病人体内ＩＬ－４水平升高可以诱导淋巴细胞表面ＦｃεＲⅡ／ＣＤ２３分子表达的增强，并在ＩｇＥ抗体形成中起着重要作用。     

弓形虫IgM抗体检测方法的研究

本文以鼠抗人μ链单克隆抗体捕获被测人血清中ＩｇＭ抗体，再以辣根过氧化物酶标记的弓形虫抗原进行直接酶联免疫吸附试验，检测人血清中特异性抗弓形虫ＩｇＭ抗体，并以阻断试验证实该方法的特异性。与风疹病毒ＩｇＭ抗体阳性血清，巨细胞病毒ＩｇＭ抗体阳性血清和类风湿因子等无交叉反应。与进口试剂盒以酶联免疫吸附试验双夹心法（ＤＳ－ＥＬＩＳＡ－Ｔｏｘ－ＩｇＭ）进行比较检测了临床血清样品１０５３份，两者阳性符合率为９５．５６％，ＤＳ－ＥＬＩＳＡ阴性血清在Ｄ－ＥＬＩＳＡ法亦阴性，而且后者灵敏度略高。酶标记抗原和包被抗体的酶标板在４℃中保存６个月仍稳定。试剂盒操作简便，快速，适用于弓形虫病的早期诊断。     

毛细支气管炎细胞因子与发病机理的研究

为了解毛支发病机理与机体免疫功能的关系,探讨环境与遗传因素对发病的影响,对３０例不同病毒感染的毛支患儿取血清及外周血单个核细胞（ＰＢＭＣ）培养,用ＥＬＩＳＡ方法测定培养上清中细胞因子及血清ＩｇＥ浓度。结果表明：①毛支急性期ＩＦＮ－γ水平降低,ＩＬ－４、ＩＬ－１０及血清ＩｇＥ水平增高。②毛支ＩＬ－４、ＩＬ－１０、ＩｇＥ水平增高与感染病毒的种类关系不明显,与机体遗传特应质有关。提示：毛支患儿存在细胞免     

肾病综合征患者PBMC上清IL—4和IFN—γ及血清IgE水平

本文报道，对４２例原发性单纯型肾病综合征（ＩＮＳ）患者及相应年龄组健康儿童进行ＩＬ－４，ＩＦＮ－γ及血清ＩｇＥ水平均显著高于正常对照组，且两者呈正相关，而ＩＦＮ－γ水平显著低于正常对照组；缓解期患者ＩＬ－４、ＩＦＮ－γ及ＩｇＥ均接近于正常水平，提示ＩＬ－４及ＩＦＮ－γ为ＩｇＥ异常变化的两个重要细胞因子，对ＩｇＥ的合成，ＩＬ－４及ＩＦＮ－γ起着正负反馈的调节作用。     

IFN—γ和IL—4对IgE生成的调节作用探讨

本文通过对鸡蛋过敏患者血清及外周血单个核细胞（ＰＢＭＣｓ）培养上清的ＩｇＥ水平测定，探讨了γ－干扰素（ＩＦＮ－γ）、白细胞介素４（ＩＬ－４）参与ＩｇＥ生成的调节机制。结果表明：ＩＦＮ－γ可抑制ＩｇＥ的生成，而ＩＬ－４则相反。ＰＢＭＣｓ培养上清中ＩＦＮ－γ和ＩＬ－４与ＩｇＥ浓度之间呈显著相关。     

IFN－γ和IL－4对IgE生成的调节作用探讨

本文通过对鸡蛋过敏患者血清及外周血单个核细胞（ＰＢＭＣｓ）培养上清的ＩｇＥ水平测定，探讨了γ－干扰素（ＩＦＮ－γ）、白细胞介素４（ＩＬ－４）参与ＩｇＥ生成的调节机制。结果表明：ＩＦＮ－γ可抑制ＩｇＥ的生成，而ＩＬ－４则相反。ＰＢＭＣｓ培养上清中ＩＦＮ－γ和ＩＬ－４与ＩｇＥ浓度之间呈显著相关。     

IL－2R水平在骨髓增生异常综合征中的意义

为了对ＭＤＳ的发生发展和免疫学异常进一步了解，我们用双抗体夹心ＥＬＩＳＡ法检测２０例ＭＤＳ患者血清中ｓＩＬ－２Ｒ水平；采用ＡＰＡＡＰ桥联酶免疫染色观察９例ＭＤＳ患者ＰＢＭＣ中ｍＩＬ－２Ｒ的表达，发现ＭＤＳ患者血清中ｓＩＬ－２Ｒ较正常人增高。在ＭＤＳ亚型中ＲＡＥＢ，ＲＡＥＢ－ｔ组较ＲＡ组增高显著，Ｐ＜０．０１；校再障组也增高。ｍＩＬ－２Ｒ阳性细胞百分率也较正常人高。血清中ｓＩＬ－２Ｒ释放水平与ＰＢＭＣ中ｍＩＬ－２Ｒ的表达比较，两者无明显相关。研究结果表明，ＭＤＳ除髓系细胞累及外，ＩＬ－２Ｒ水平增高可能是ＭＤＳ淋巴细胞异常，免疫系统功能紊乱的一种表现。     

Candida albicans mannan- and protein-induced humoral, cellular and cytokine responses in atopic dermatitis patients

BACKGROUND: The cytokine observed most often in atopic dermatitis (AD) is IL-4, but a role for IL-5 and IFN-gamma in the late and delayed phase reactions has been suggested. In AD with head, neck and shoulder distribution, hypersensitivity to saprophytic yeasts is an important pathogenetic factor. The yeast allergens include both the mannan polysaccharides and the proteins. Mannans are major cross-reacting allergens likely to be involved in the pathogenesis of AD. OBJECTIVE: To characterize the humoral, lymphoproliferative and cytokine (IL-2, 4, 5 and IFN-gamma) responses of peripheral blood mononuclear cells (PBMCs) induced by Candida albicans mannan and protein antigens in AD. METHODS: Fifteen AD patients and seven healthy controls were included. Ficoll-isolated PBMCs were stimulated by PHA and laboratory-generated mannan and protein extracts of C. albicans. Lymphocyte proliferation was measured and cytokine production was studied by ELISA. The antigen-specific IgG and IgE antibodies were analysed by ELISA and nitrocellulose RAST. RESULTS: In AD mannan (P < 0.005) and protein (P < 0.002), specific IgE levels were higher than in healthy controls. Both mannan and protein-specific lymphoproliferations (both: P < 0.02) were higher in AD than in healthy controls. Mannan, but not protein, induced long lasting IL-2 and IL-4 productions from 24 h lasting up to 66-96 h and IL-5 and IFN-gamma productions with elevated levels at 66 and 96 h. The mannan-induced IL-2 (P = 0.015) and IFN-gamma (P < 0.005) were increased in AD as compared with healthy controls. Significant correlations were seen between the protein-induced proliferation responses and both serum total IgE (r = 0.59, P < 0.01) and protein-specific IgE (r = 0.65, P < 0.005). The mannan-induced IL-2 responses correlated with the specific IgE (r = 0.62, P < 0.01) and proliferation (r = 0.51, P < 0.02) and S-IgE level (r = 0.71, P < 0. 002). Mannan-induced IL-4 and IFN-gamma productions also correlated (r = 0.43, P < 0.05). CONCLUSIONS: C. albicans mannan induced elevated IL-2 and IFN-gamma responses in AD patients. The correlations of the cytokine responses with mannan-induced IgE and proliferation responses suggest that C. albicans mannan induced TH1 type cytokine responses are involved in AD.     

Association of the single-nucleotide polymorphism and haplotype of the interleukin 18 gene with atopic dermatitis in Koreans

BACKGROUND: IL-18 is a proinflammatory cytokine that plays an important role for T-helper type 1 (Th1) and Th2 cytokine in the presence/absence of IL-12. It has been recently shown that human IL-18 plays a role in atopic dermatitis (AD) by enhancing IL-4 and IL-13 production and by stimulating the synthesis of IgE. OBJECTIVE: We wanted to evaluate the associations of single-nucleotide polymorphism (SNP) and the haplotype in the IL-18 gene, hence we performed genotyping for the SNPs in the IL-18 gene in AD patients and normal controls. METHOD: We genotyped three SNPs from the IL-18 gene for the 1120 case-control samples (646 AD patients and 474 normal controls). We measured the serum IL-18, IL-4 and IL-13 concentrations in 74 individuals (25 ADe, 25 ADi and 24 controls) by performing ELISA. RESULT: The rs795467 SNP and haplotype T-T-C were significantly associated with AD, and especially between the ADe and normal control groups (P=0.03 and 0.01). The serum IL-18 concentration was higher in the AD group than in the normal controls without any correlation with the rs795467 polymorphism. We did not find any correlations between the serum IL-18 levels and the SCORing atopic dermatitis index, the blood eosinophil counts and the ECP, and there was no correlation between the serum IL-18 levels and the serum IL-4 and IL-13 levels. CONCLUSION: We found that the one SNP and the haplotype T-T-C were strongly associated with the allergic type of AD, but not with the non-allergic intrinsic type. These data support the hypothesis that IL-18 up-regulates IgE production, yet more experiments will be needed to prove the in vivo involvement of Th2 cytokine.     

High serum levels of additional IL-18 forms may be reciprocally correlated with IgE levels in patients with atopic dermatitis

We established an ELISA system for determination of as yet unidentified species of interleukin 18 (IL-18), named IL-18 type 2, in human serum. Serum IL-18 levels and their effect on IgE levels were examined in 18 patients with atopic dermatitis (AD) with no other allergic symptoms. Three of these patients showed high IL-18 type 2 concentrations (25-100 ng/ml) in their blood serum, and this IL-18 type 2 was detectable only with our established ELISA system. In contrast, the level of the conventional form of IL-18 (type 1) was found to be 50-400 pg/ml in all patients by the commercially available ELISA. The levels of type 1 IL-18 showed no correlation with those of type 2 and approximately 2-fold higher in AD patients than in normal subjects. IL-12 p40 and IgE levels were correlated in the patients with no IL-18 type 2, and interestingly, relatively low IgE concentrations were detected in the three IL-18 type 2-positive patients. They showed considerable levels of IL-12 p40 unlike normal subjects. The IFNgamma-inducing activity of IL-18 type 2 was >100-fold less potent by weight ratio than that of a recombinant 'active' IL-18 preparation, even after the treatment with Caspase 1. Although the relationship between AD and serum IgE levels is not clear cut, IL-18 type 2 appears to play some roles in the Th2-polarization involving IgE production in association with immune responses occurring in local inflammatory milieu such as atopic lesions.     

Interleukin-18 is elevated in the sera from patients with atopic dermatitis and from atopic dermatitis model mice, NC/Nga

BACKGROUND: Several lines of in vitro and in vivo studies have demonstrated that interleukin-18 (IL-18) shows both antiallergic and allergy-promoting activities. But its expression in allergic diseases remains unknown. METHODS: Serum IL-18 levels from atopic dermatitis (AD) model mice, NC/Nga and control mice and from patients with AD and healthy volunteers were measured by ELISA. The relationship between IL-18 levels and serum IgE levels or clinical severity was also examined. RESULTS: Serum IL-18 levels from NC/Nga mice were significantly increased compared to those from control mice. The elevation of IL-18 in the sera was observed prior to the onset and during the development of dermatitis in NC/Nga mice. In addition, IL-18 levels in the sera from patients with AD were significantly (p < 0.05) elevated compared to those from healthy volunteers. However, serum IL-18 levels tended to correlate negatively with serum IgE levels in patients with AD and NC/Nga mice. CONCLUSION: IL-18 is overexpressed in AD.     

29例特应性皮炎患者血清总IgE水平的检测

文本用双抗体夹心酶联免疫吸附试验(ELISA)测定了21例无过敏史和寄生虫卵检查阴性的健康对照及29例临床确诊为特应性皮炎(AD)患者的血清总IgE水平。结果表明:(1)AD病人血清总IgE水平明显高于正常对照,经统计学处理差别非常显著(P<0.01);(2)IgE水平与临床症状有呈正相关的倾向,症状严重者IgE水平较高,反之较低,但其差别未达到统计学上有意义的程度;(3)IgE水平与家族过敏史的相关性不明显,具有家族过敏史的AD病人其血清总IgE水平较无家族史者高,但无统计学意义;IgE水平与个人过敏史的关系亦不明显;(4)並非所有AD患者的血清总IgE水平都高于对照组的几何均值,对其可能的原因进行了分析。     

Pityrosporum and Candida specific and non-specific humoral, cellular and cytokine responses in atopic dermatitis patients

BACKGROUND: Recent cytokine (RT-PCR, ELISA) analyses of inflammation in atopic dermatitis (AD) have suggested a role for IL-4, IL-5 and IFNgamma. Pityrosporum ovale and Candida albicans are important allergens in some patients with AD of the seborrhoic head, neck and shoulder region. In AD patients, the saprophytic yeasts induce IgE responses while they usually induce TH1 type responses. The cytokine responses induced by yeasts in AD are sparsely investigated. OBJECTIVE: To characterize the P. ovale- and C. albicans-specific and non-specific humoral, lymphoproliferative and cytokine (IL-2, 4, 5 and IFNgamma) responses in AD. METHODS: Fifteen AD patients and seven healthy controls (HC) were included. Ficoll-isolated PBMC were stimulated by PHA and laboratory-generated extracts of P. ovale and C. albicans. Lymphocyte proliferation was measured by 3H-thymidine incorporation and cytokine production by sandwich-ELISAs. The antigen-specific IgG and IgE antibodies were analysed by ELISA and nitrocellulose RAST. RESULTS: Pityrosporum ovale- and C. albicans-specific IgE (both P < 0.001) and P. ovale-induced PBMC proliferation (P < 0.02) were elevated in AD. In general, the IL-4/IFNgamma ratio induced by P. ovale was higher than that induced by C. albicans (P < 0.01). The PHA-induced IL-2 (P < 0.05) and IL-4 responses (P < 0.005), and the C. albicans-induced IL-5 response (P < 0.02) and IFNgamma response (P < 0.01), were elevated in AD. A network of correlations was seen between serum total and the yeast-specific IgE, P. ovale-specific lymphoproliferation, PHA-induced IL-2, IL-4 and IL-5, and C. albicans-induced IL-5. CONCLUSION: The cytokine profiles found in this study support the role of TH0 or TH1 cells by the side of TH2 cells in the pathogenesis of atopic dermatitis. Pityrosporum ovale appears to be associated more with IL-4 responses and C. albicans with IFNgamma responses.     

An obligate role for T-cell receptor alphabeta+ T cells but not T-cell receptor gammadelta+ T cells, B cells, or CD40/CD40L interactions in a mouse model of atopic dermatitis

BACKGROUND: We recently described a murine model of atopic dermatitis (AD) elicited by epicutaneous sensitization with ovalbumin (OVA). The skin lesions in these mice were characterized by a dermal infiltrate consisting of eosinophils and T cells and by increased expression of the TH2 cytokines IL-4 and IL-5. Epicutaneous sensitization induces a rise in the levels of serum total IgE and OVA-specific antibodies, further indicating that it elicits a predominantly TH2 response. OBJECTIVE: This study was undertaken to assess the roles of T cells, B cells, and CD40L-CD40 interactions in AD. METHODS: Mice with targeted gene deletions were sensitized with OVA. Histologic and immunohistochemical examinations, as well as measurements of IL-4 mRNA, were performed on OVA-sensitized skin. Total and antigen-specific serum IgE levels were determined. RESULTS: RAG2(-/-) mice, which lack both T and B cells, did not exhibit cellular infiltration, induction of dermal IL-4 mRNA, or elevation of serum IgE after OVA sensitization; all of these features were present in B-cell-deficient IgH(-/-) mice. T-cell receptor alpha(-/-) mice did not display cellular infiltration, IL-4 mRNA expression, or increased IgE levels after OVA sensitization, but these responses were elicited in T-cell receptor delta(-/-) mice after sensitization. Absence of CD40 had no effect on these responses. CONCLUSION: These results suggest that alphabeta T cells, but not gammadelta T cells, B cells, or CD40L-CD40 interactions, are critical for skin inflammation and the TH2 response in AD.     

Effect of Chrysanthemi borealis flos on atopic dermatitis induced by 1-chloro 2,4-dinitrobenzene in NC/Nga mouse

The flower of Chrysanthemum boreale has traditionally been used for treatment of various inflammatory disease including atopic dermatitis (AD). However, its action on AD is unclear. Therefore, we investigated the effect of CB on AD using NC/Nga mice as an AD model. The effect of CB on 1-Chloro-2,4-dinitrobenzene (DNCB) induced NC/Nga mice was evaluated by examining skin symptom severity, itching behavior, ear thickness, levels of serum Immunoglobulin E (IgE), tumor necrosis factor-α (TNF-α), and interleukin-4 (IL-4), skin histology. The CB significantly reduced the total clinical severity score, itching behavior, ear thickness and the level of serum IgE in AD mouse model. CB not only decreased TNF-α but also IL-4. These results suggest that CB may be a potential therapeutic modality for AD.     

Effect of Chrysanthemi borealis flos on atopic dermatitis induced by 1-chloro 2,4-dinitrobenzene in NC/Nga mouse

The flower of Chrysanthemum boreale has traditionally been used for treatment of various inflammatory disease including atopic dermatitis (AD). However, its action on AD is unclear. Therefore, we investigated the effect of CB on AD using NC/Nga mice as an AD model. The effect of CB on 1-Chloro-2,4-dinitrobenzene (DNCB) induced NC/Nga mice was evaluated by examining skin symptom severity, itching behavior, ear thickness, levels of serum Immunoglobulin E (IgE), tumor necrosis factor-α (TNF-α), and interleukin-4 (IL-4), skin histology. The CB significantly reduced the total clinical severity score, itching behavior, ear thickness and the level of serum IgE in AD mouse model. CB not only decreased TNF-α but also IL-4. These results suggest that CB may be a potential therapeutic modality for AD.     

Interleukin-10 haplotype associated with total serum IgE in atopic dermatitis patients

BACKGROUND: The genetic background of atopic dermatitis (AD) is not clearly understood. Interleukin (IL)-10 is a powerful Th-2 cell cytokine produced by lymphoid cells that exerts its function by inhibiting macrophage/monocyte and T-cell lymphocyte replication and secretion of inflammatory cytokines [IL-1, tumour necrosis factor-alpha (TNFA), IL-6, IL-8 and IL-12]. OBJECTIVE: In an effort to discover additional polymorphism(s) in genes whose variant(s) have been implicated in total immunoglobulin E (IgE) level in AD patients, we scrutinized the single nucleotide polymorphisms (SNPs) in the IL10 gene as a potent candidate for contributing to the level of IgE in serum. METHODS: We recruited 334 AD patients and assayed their serum total IgE levels using the LIPA-200 system. Four SNPs in the IL10 gene were genotyped using the single-base extension (SBE) method. Logistic regression analyses were performed with single polymorphisms and haplotypes (ht) to determine their association with the level of serum total IgE. RESULTS: Genetic association analysis of total serum IgE in AD patients revealed that one of the IL10 ht, IL10-ht2, was associated with decreased serum total IgE in gene dose-dependent manner (P = 0.02-0.001). CONCLUSIONS: It was predicted that the inhibition of innate immunity by increased IL-10 production in IL10-ht2-bearing individuals might be associated with decreased total serum IgE levels among AD patients. The greater effects of IL10 ht on decreased total serum IgE levels suggest that the effect of IL-10 polymorphism might be the result of a combined genotype (ht) rather than single polymorphisms.