FcεR I-β基因启动子区多态性与哮喘儿童血清总IgE的关系

目的　探讨IgE高亲和力受体 β链 (FcεRI β)基因启动子区 10 9C/T多态性与儿童哮喘血清总IgE水平的关系。方法　采用聚合酶链式反应 限制性片断长度多态性 (PCR RFLP)法检测了 2 0 0 1年 12月至 2 0 0 3年 2月吉林大学第一临床医院 12 6例哮喘儿童和 87名正常儿童的FcεRI β 10 9C/T基因多态性 ;用酶联免疫法检测血清总IgE水平和空气过敏原。结果　FcεRI β 10 9C/T基因型分布及等位基因频率哮喘组与正常对照组比较 ,差异均无显著性意义 ;哮喘组、对照组过敏原检测阳性率在突变基因型与未突变基因型间比较 ,差异均无显著性意义 ;哮喘组FcεRI β 10 9C/T突变纯合型个体的Log10 IgE水平高于突变杂合型及未突变型个体的水平 (P <0 0 5 )。结论　FcεRI β基因启动子区 10 9C/T不是汉族儿童哮喘的易感基因 ,与过敏原亦无关 ,但突变纯合子TT型与血清总IgE水平升高相关联。     

白细胞介素-4基因启动子多态性与广西壮族支气管哮喘患儿易感性和血清总免疫球蛋白E水平的相关性

目的研究IL-4基因启动子区域-589C/T和-33C/T位点多态性在广西壮族儿童中的分布及与壮族儿童支气管哮喘(哮喘)易感性及血清总IgE水平的相关性。方法采用聚合酶链反应-限制性片段长度多态性方法对健康儿童102例和哮喘患儿72例IL-4基因-589位点和-33位点进行分析,ELISA方法检测2组血清总IgE水平。采用SPSS 14.0软件进行统计学分析。结果1.IL-4-589位点在健康对照组中基因型分布频率为CC 5.9%、CT 23.5%、TT 70.6%,哮喘组为CC 2.8%、CT 19.4%、TT 77.8%;健康对照组等位基因频率为C 17.6%、T 82.4%,哮喘组为C 12.5%、T 87.5%。2.IL-4-33位点在健康对照组和哮喘组中基因型分布和等位基因频率与IL-4-589位点频率分布一致,连锁不平衡值△=0.978。3.哮喘组与健康对照组之间基因型频率和等位基因频率比较差异均无统计学意义(Pa>0.05)。4.哮喘组血清总IgE水平较健康对照组明显增高(P<0.01);各组不同基因型间血清总IgE水平比较差异均无统计学意义(Pa>0.05);3种相同基因型不同组间比较哮喘组血清总IgE水平均较健康对照组高(Pa<0.05)。结论在广西地区壮族儿童人群中,IL-4基因启动子-589位点和-33位点存在多态性,两位点存在连锁不平衡,其多态性与壮族儿童哮喘的易感性无关联,与血清总IgE水平无相关性。     

哮喘儿童白细胞介素13基因型与表型的关联研究

目的探讨哮喘儿童白细胞介素13(IL-13)Arg130Gln基因型与血清总IgE、IL-13水平及过敏原表型之间的关系。方法采用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)法,对130例哮喘患儿和100例健康对照儿童进行IL-13Arg130Gln基因多态性的检测,并用酶免法检测血清总IgE、IL-13水平和空气过敏原。结果基因型分布及等位基因频率哮喘组与对照组比较差异均无显著性,哮喘组突变型个体的Log10IL-13水平高于未突变型个体的水平(P<0.05),哮喘组突变纯合型个体的Log10IgE水平高于突变杂合型及未突变型个体的水平(P<0.05),哮喘组及对照组突变型与未突变型的过敏原阳性率比较差异均无显著性。结论IL-13Arg130Gln突变不是中国长春地区儿童哮喘的易感基因,但此突变可能与血清IL-13蛋白质水平升高有关,突变纯合型可能与血清总IgE水平相关联。     

转化生长因子β1基因多态性与儿童哮喘易感性研究

目的探讨转化生长因子β1(TGFβ1)基因多态性与儿童哮喘的关系。方法应用聚合酶链反应 -限制性片段长度多态性 (PCR_RFLP)技术对98例哮喘患儿与52例正常儿童进行TGFβ1 基因5′-侧翼区 -509C/T多态性分析 ,并采用双抗体夹心ELISA法检测血清总IgE水平。结果哮喘组基因型分布和等位基因频率与对照组相比 ,差异均无显著性 (P>0.05) ;重度哮喘组基因型分布和等位基因频率与轻度组或对照组相比 ,差异有显著性(P<0.05) ;基因型为TT型的哮喘患儿血清总IgE水平明显高于CC型或CT型 ,差异具有显著性 (P<0.05)。结论TGFβ1 基因 -509C/T突变不是儿童哮喘的高发因素 ,但它可能是重度哮喘的候选基因 ,且突变纯合型与血清总IgE水平升高有关。     

儿童哮喘IgE高亲和力受体β链基因多态性分析

目的 研究儿童哮喘IgE高亲和力受体β链(Fc ε RI-β)基因多态性。方法 采用扩增阻滞突变系统-聚合酶链式反应(ARMS-PCR)技术对151例哮喘儿童和。105例健康对照儿童进行Fc ε RI-β基因第6外显子区1181L、V183L和第7外显子区E237G位点的多态性检测，分析其与儿童哮喘易感性的关系，并用酶免法检测血清总IgE水平及过敏原表型，研究上述3个编码区的基因型与表型的关系。结果 1181L突变与儿童哮喘有显著相关性(OR=2．00，95％CI=1．10～3．11)，哮喘儿童及健康对照儿童均不存在V183L突变或突变率极低，E237G突变与儿童哮喘无显著相关性；1181L及E237G突变均与过敏原或血清总IgE水平增高无关。结论 Fc ε RI-β 1181L基因突变是我国长春地区儿童哮喘的易感基因，但无足够证据证明Fc ε RI-β基因是儿童哮喘特应性的决定性基因。     

白细胞介素-4基因C-589T多态性与呼吸道合胞病毒毛细支气管炎发病的相关性

目的 探讨白细胞介素-(IL-4)基因C-589T位点多态性与呼吸道合胞病毒(RSV)毛细支气管炎发病的相关性.方法 采用聚合酶链-限制性片段长度多态法(PCR-RFLP)检测RSV毛细支气管炎组(毛支组)和健康对照组儿童IL-4/C-589T位点多态性;用化学发光法和酶联免疫分析法(ELISA)分别检测RSV毛支组血清总IgE和鼻咽分泌物(NPS)中IL-4水平.结果 二组均发现IL-4/C-589T位点基因多态性,但仅见TT和CT 2种基因型,以TT纯合子基因型为主,其中RSV毛支组IL-4/C-589T位点TT、CT基因型频率分别为94.6%、5.4%,T、C等位基因频率分别为97.3%、2.7%;对照组TT、CT基因型频率分别为76.9%、23.1%,T、C等位基因频率分别为88.4%、11.6%.RSV毛支组TT基因型和T等位基因频率均显著高于对照组(χ2=15.995,14.842 Pa＜0.01),且T等位基因携带者患病的危险性为C等位基因携带者的4.73倍(OR=4.73 P=0).但2种基因型RSV毛支患儿间NPS中IL-4及血清总IgE水平均无显著性差异(Z=0.515,t=0.260 Pa＞0.05).IL-4/C-589T位点多态性在轻度和中重度患儿间无显著差异(χ2=0.024 P＞0.05).结论 温州地区儿童存在IL-4/C-589T位点的多态性.IL-4/C-589T 位点T等位基因可能是影响RSV毛细支气管炎发病的一个重要候选基因.     

CTLA-4启动子-318C/T基因多态性与哮喘患儿血清总IgE水平的关系

目的探讨细胞毒T淋巴细胞相关抗原4(CTLA-4)启动子-318(C/T)基因多态性和吉林长春地区哮喘患儿血清总IgE水平的关系,以便为哮喘的诊治提供新线索。方法随机选取90例哮喘患儿及100例健康体检儿童作对照组,哮喘组和对照组的性别和年龄差异无统计学意义。采用聚合酶链式反应—限制性片段长度多态性(PCR-RFLP)技术对哮喘组和对照组CTLA-4启动子-318(C/T)基因多态性进行检测分析;同时采用酶联免疫吸附试验(ELISA)检测不同基因型哮喘患儿血清总IgE水平。结果CTLA-4启动子-318位点存在基因多态性。血清总IgE水平哮喘组(M=308.92 IU/ml,Q=254.75)高于正常对照组(M=36.66 IU/ml,Q=33.57),P<0.01;哮喘组中基因型为野生型(CC型,M=375.86 IU/ml,Q=139.95),高于突变型(TT型 CT型,M=141.25 IU/ml,Q=47.73),P<0.01。结论CTLA-4启动子-318(C/T)存在基因多态性,该位点的基因多态性可能引起血清总IgE水平下调。     

哮喘儿童IgE高亲合力受体β链基因多态性的初步研究

目的：探讨IgE高亲合力受体β链基因的I181L,V183L和E237G位点基因多态性与儿童哮喘及血浆总IgE的关系。方法：采用扩增阻滞突变系统聚合酶链式反应(ARMS-PCR)检测技术对50例四川地区哮喘儿童和40例健康对照儿童进行181,183,237三个位点的多态性进行检测,并用双抗体夹心ELISA法检测血清总IgE水平,分析其与儿童哮喘易感性的关系。结果：在哮喘儿童中检测到I181L突变5例、V183L突变2例、E237G突变7例,在对照组中未发现突变子;有突变的哮喘儿童的血清IgE明显高于无突变和正常对照儿童。I181L和E237G突变与儿童哮喘和血清IgE升高有显著相关性。结论：FcεR1β基因I181L和E237G基因多态性是儿童哮喘发病的重要候选基因,与血清总IgE水平升高密切相关。     

哮喘患儿白细胞介素13、γ-干扰素、免疫球蛋白E的变化及过敏原检测

目的 探讨哮喘患儿白细胞介素-13(IL-13)、γ-干扰素(IFN-γ)及免疫球蛋白E(IgE)的变化，了解长春市儿童哮喘常见过敏原。方法 用双抗体夹心ELISA法检测50例哮喘患儿血清IL-13、IFN-γ及IgE水平，并分析三者之间关系；用酶免法进行过敏原特异性IgE(sIgE)定性检测。结果 血清IL-13及IgE水平发作组显著高于缓解组，发作组与缓解组均显著高于正常对照组。血清IFN-γ水平发作组与缓解组均显著低于正常对照组。直线相关分析表明，哮喘患儿血清IL-13水平与IgE呈正相关，IFN-γ水平与IgE、IL-13均呈负相关。屋尘和尘螨是长春市儿童哮喘主要过敏原。结论哮喘患儿发作期及缓解期均存在IgE、IL-13和IFN-γ水平失衡。     

白介素-4基因C-33T多态性与呼吸道合胞病毒毛细支气管炎的相关性

目的 研究白介素-4(IL-4)基因C-33T与呼吸道合胞病毒(RSV)毛细支气管炎的易感性、病情严重程度的关系及对血清总IgE和鼻咽分泌物(NPS)IL-4水平的影响.方法 采用聚合酶链-限制性片段长度多态法(PCR-RFLP)检测130例RSV毛细支气管炎患儿和108例对照组儿童IL-4/C-33T位点多态性,分别用化学发光法和酶联免疫分析法,检测RSV毛细支气管炎患儿血清总IgE和NPs中IL-4水平.结果 两组IL-4启动子区C-33T位点均可见TT、CT和CC 3种基因型,其中病例组,TT、CT和CC基因型频率分别为66.9%、26.9%和6.2%,对照组分别为69.4%、26.9%和3.7%,两组差异无统计学意义(X2=0.758,P>0.05);病例组T、C等位基因频率分别为80.3%、19.7%,对照组分别为82.9%、17.1%,两组差异亦无统计学意义(X2=0.073,P>0.05;OR=0.847,P>0.05).病例组三种基因型间NPS中IL-4及血清总IgE水平差异均无统计学意义(H=0.103,F=0.529,P均>0.05);三种基因型频率在轻度组和中重度组间的差异亦无统计学意义(X2=0.825,P>0.05).结论 温州地区儿童存在IL-4/C-33T位点的多态性,但未发现其与RSV毛细支气管炎存在关联.     

支气管哮喘患儿亚甲基四氢叶酸还原酶基因多态性与血清免疫球蛋白E的关系

目的探讨支气管哮喘(哮喘)患儿亚甲基四氢叶酸还原酶(MTHFR)基因C677位点多态性与血清IgE水平的相关性。方法选择95例哮喘患儿作为病例组,均为急性发作期或临床缓解期哮喘患儿,患病前2周均未使用过肾上腺皮质激素及免疫调节剂。另选择健康儿童113例作为健康对照组。2组儿童年龄及性别比较差异均无统计学意义。采用PCR-限制性片段长度多态性分析法对病例组和健康对照组儿童外周血白细胞MTHFR基因C677位点基因多态性进行研究,应用双抗体夹心ELISA法检测2组儿童血清总IgE水平。结果健康对照组MTHFR 677C/T的3种基因型频率分别CC 35.4%、CT 45.1%、TT 19.5%,病例组分别为CC 24.2%、CT 40.0%、TT 35.8%,677C/T基因型分布频率在哮喘病例组和健康对照组间差异有统计学意义(χ2=7.556 5,P<0.05)。病例组T等位基因的频率为55.3%,健康对照组为42.0%,病例组较健康对照组显著增高,其患哮喘的危险度是健康对照组的1.71倍(χ2=7.254 7,P<0.01;95%CI:1.13～2.57)。病例组血清总IgE水平在各基因型患儿间比较差异有统计学意义(F=3.46,P<0.05),健康对照组血清总IgE水平在各基因型儿童间比较差异无统计学意义(F=0.13,P>0.05)。结论 MTHFRC677位点C→T的基因突变增加了患儿哮喘发病的危险度,TT基因型与哮喘的发生直接相关;哮喘患儿血清总IgE水平升高,该位点基因型并非导致血清总IgE水平升高的直接原因。     

The effects of NOS1 gene on asthma and total IgE levels in Taiwanese children,and the interactions with environmental factors

Wang T‐N, Tseng H‐I, Kao C‐C, Chu Y‐T, Chen W‐Y, Wu P‐F, Lee C‐H, Ko Y‐C. The effects of NOS1 gene on asthma and total IgE levels in Taiwanese children, and the interactions with environmental factors.
Pediatr Allergy Immunol 2010: 21: 1064–1071.
© 2010 John Wiley & Sons A/S Asthma is a complex disorder, which is known to be affected by interactions between genetic and environmental factors. The aim of this study was to investigate the three microsatellite polymorphisms of GT repeats in intron 2, AAT repeats in intron 20, and CA repeats in exon 29 of the NOS1 gene in 155 asthmatic children and 301 control children, and the interaction with environmental factors in southern Taiwan. Total serum IgE, phadiatop test and genetic polymorphisms were measured. The genotype frequency of 14/14‐AAT repeats of the NOS1 gene was significantly higher in the asthmatic group (p = 0.01). Total IgE concentrations were higher in asthmatic children (p = 0.015) carrying the NOS1 14/14‐AAT genotype than in subjects with other polymorphisms. The gene and environmental interaction effects were 3.83‐fold, 6.86‐fold, and 8.04‐fold (all corrected p‐values <0.001) between subjects carrying at least one NOS1 14‐AAT allele and exposure to cockroaches, high levels of total IgE, and positive response against the phadiatop test in asthmatic children. The findings of this study provide strong evidence that NOS1 gene with 14‐AAT tandem repeats has a significant effect in asthmatic children. Environmental factors and atopic status will enhance the asthmatic risk for children who carry NOS1 susceptible allele.     

198例哮喘儿童遗传易感性研究

目的：分析4种哮喘相关基因位点在198例哮喘患儿中的单核苷酸多态性（single nucleotide polymorphismr,SNP）分布特点,并探讨其与儿童哮喘遗传易感性及某些哮喘临床表型的关系。方法：采用聚合酶链反应产物电泳鉴定法及实时荧光定量核酸扩增技术检测4种哮喘相关基因位点SNP在198名哮喘患儿和110名健康对照者中的分布,并测定哮喘患儿的血清总IgE（TIgE）水平及血嗜酸性粒细胞比例（%EOS）,同时对4种哮喘相关基因位点不同基因型间的TIgE及%EOS分别进行比较分析。结果：血管紧张素转换酶（ACE）基因位点纯合子基因型（DD）频率在哮喘组明显高于健康对照组（χ2=30.667,P＜0.01）,且D型等位基因频率在哮喘组亦明显高于健康对照组（χ2=7.151,P<0.01）;各基因位点多态性分布与血清TIgE水平及%EOS均无相关性（均P＞0.05）。结论：ACE基因DD基因型与儿童哮喘的遗传易感性有关,可能是儿童哮喘的危险因素。     

Cytokine production, activation marker, and skin homing receptor in children with atopic dermatitis and bronchial asthma

T cells are known to develop a critical role in the pathogenesis of atopic dermatitis (AD) and bronchial asthma. T cells involved in AD express the skin homing receptor CLA, but no lung homing receptor has been identified in bronchial asthma. We compared different cell markers and the cytokine production in T cells from children with AD or bronchial asthma. We studied the involvement of CLA+ and CLA- T-cell subpopulations in these diseases. We studied 20 children with acute AD lesions, 15 with mild persistent asthma, and 15 non-atopic controls. All patients were sensitized to house dust mite (DP) and evaluated during the acute phase. Total and specific IgE were measured by immunoassay and the expression of different cell markers and the cytokine production was analyzed by flow cytometry in peripheral blood mononuclear cells. Total IgE was significantly higher in AD children and IgE to DP in the asthmatic children. There was a significant increase in CD25+ CD4+ cells in asthmatic children and in HLA-DR+ CD4+ and HLA-DR+ CD8+ cells in AD. In the CD4+ subsets, there was an increase in IL-13, IL-5 and TNF-alpha in AD compared to controls, a decrease in IFN-gamma in asthmatic children compared to controls, and an increase in IL-13, IL5, IL2, TNF-alpha, and IFN-gamma in the AD compared to asthmatic children. Changes in cytokine production were mainly detected in CLA+ cells in AD and in CLA- cells in asthma. Differences exist in total and specific IgE, activation markers, and cytokine patterns between AD children and children with asthma, with the former expressing a Th2 pattern whereas in asthmatic children we only detected a decrease in IFN-gamma. Moreover, the subpopulations (CLA+ vs. CLA-) expressing these changes were different, indicating that the underlying mechanisms in the two diseases are not exactly the same.     

CD4+CD25(high) Treg cells in peripheral blood during remission and exacerbation of allergic asthma in children

Aim: To determine the percentage of CD4⁺CD25^high Treg cells in peripheral blood CD4⁺ T cells of allergic asthmatic children during disease remission and exacerbation. Methods: Peripheral blood mononuclear cells (PBMC) and serum samples were collected from 6‐ to 11‐year‐old children with mild‐to‐moderate allergic asthma (n = 34) and from healthy controls (n = 15). CD4⁺CD25^high T cells in PBMC were detected by flow cytometry. Total and specific IgE in serum were analysed by enzyme‐amplified chemiluminescence, and IL‐2 was measured by ELISA. Results: There was no significant difference in CD4⁺CD25^high T‐cell proportions between asthmatic children in exacerbation and remission as compared with controls. CD4⁺CD25^high T‐cell percentages were not correlated with total and specific IgE. IL‐2 was elevated in both disease remission and exacerbation but did not correlate significantly with CD4⁺CD25^high T‐cell percentages. Conclusion: CD4⁺CD25^high T‐cell proportion in the peripheral blood of total CD4⁺ T cells is not reduced in children with allergic IgE‐mediated asthma and does not differ between disease remission and exacerbation.     

哮喘缓解期儿童血清白细胞介素-13与免疫球蛋白E水平变化

目的探讨哮喘患儿IL-13及总IgE变化的意义。方法采用酶联免疫吸附试验(ELISA)方法和Pharmacia UniCAP检测系统检测88例6~14岁的哮喘缓解期和40例正常对照儿童血清IL-13与总IgE水平。结果哮喘缓解期儿童临床症状和肺功能改善;血清IL-13水平高于正常儿童对照组(U=3.93 P<0.01);血清总IgE水平仍明显高于正常儿童对照组(U=10.52 P<0.01)。血清IL-13水平与总IgE水平呈显著正相关(r=0.2685 P>0.05)。结论哮喘缓解期变态反应性炎症持续存在,血清高质量浓度的IL-13和IgE与哮喘的免疫病理相关。     

IL13 variants are associated with total serum IgE and early sensitization to food allergens in children with atopic dermatitis

Increased total and specific serum immunoglobulin E (IgE) levels are common characteristics of atopic diseases and their basal production is proposed to be under strong genetic control. Interleukin 13 (IL13) variants have been consistently associated with total serum IgE levels in white populations with a strongest association in non‐atopics. The aim of this study was to test the IL13 p.R130Q and c.1‐1111C>T variants in children with atopic dermatitis (AD) for associations with total serum IgE and early sensitization to common food and inhalant allergens and with asthma. We included 453 children with AD [participants of the Early Treatment of the Atopic Child (ETAC) study] that were followed from the age of 12–24 months for 3 yr. Total and specific IgE were determined at four time points. We genotyped the IL13 p.R130Q and c.1‐1111C>T variants by melting curve analysis. In children up to 4 yr of age, the 130Q allele was related to slightly higher total IgE levels compared to heterozygotes and 130R homozygotes. More importantly, both IL13 variants were significantly associated with sensitization to food allergens, with most significant results for sensitization to egg (p = 0.0001). Although early sensitization to hen’s egg represents a strong risk factor for subsequent sensitization to inhalant allergens and asthma, the investigated IL13 variants were not associated with these phenotypes at the age of 48–60 months. In summary IL13 variants contribute to elevated levels of total serum IgE in young atopic children and are strongly associated with sensitization to food allergens, particularly to hen’s egg. These findings suggest that IL13 variants play a major role not only in non‐cognate but also in allergen specific IgE synthesis.