CD4^＋T细胞与儿童免疫性疾病研究进展

CD4 T细胞是一群具有不同功能的异质性细胞。过去认为CD4 T细胞主要是辅助性T(Th)细胞,通常分为Th1型和Th2型。近来发现,机体存在一种新型CD4 效应T细胞Th17细胞。此细胞具有独立的分化和发育调节机制,并产生特征性的IL-17效应因子,在自身免疫性疾病和感染性疾病中发挥重要的调节作用。现就Th1/Th2细胞与Th17细胞的分化条件,及Th17细胞在自身免疫、机体防御与儿童免疫性疾病中的可能作用作一探讨。     

过敏性紫癜患儿血树突状细胞共刺激分子表达及其与辅助性T淋巴细胞1/辅助性T淋巴细胞2平衡相关性

目的观察过敏性紫癜(HSP)患儿外周血树突状细胞(DC)共刺激分子表达及其与辅助性T淋巴细胞(Th)1/Th2平衡的相关性。方法HSP患儿40例。健康对照儿童18例。酶联免疫吸附法(ELISA)检测其血浆IFNγ-、IL-4水平。对其中HSP 28例及18例健康对照儿童外周血单个核细胞(PBMC)体外经细胞因子重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)、rhIL-4和rhTNF-α以诱生DC并培养8 d,流式细胞仪检测DC表面CD86(B7-2)、CD80(B7-1)、CD83表达率。结果1.HSP组血浆IFN-γ水平显著低于健康对照组(t=4.26 P<0.01),IL-4水平显著高于健康对照组(t′=5.09 P<0.01),IFN-γ/IL-4比值显著低于健康对照组(t′=7.98 P<0.01)。2.HSP组外周血DC表面CD83表达率与健康对照组无显著性差异(P>0.05);HSP组外周血DC表面CD86表达率显著高于健康对照组(t=3.94 P<0.01),CD80低于健康对照组(t′=2.60 P<0.05)。3.二组外周血DC表面CD86表达率与血浆IL-4水平均呈显著正相关(r=0.53,0.63 Pa<0.01),与IFN-γ/IL-4比值均呈显著负相关(r=-0.55,-0.80 Pa<0.01),而与血浆IFN-γ水平均无相关性(Pa>0.05);二组外周血DC表面CD80表达率与血浆IFN-γ水平均呈正相关(r=0.43,0.49 Pa<0.05),与IFN-γ/IL-4比值均呈显著正相关(r=0.49,0.63 Pa<0.05),而与血浆IL-4水平均无相关性(Pa>0.05)。结论HSP患儿存在Th1/Th2失衡,HSP患儿DC表面共刺激分子差异表达直接或间接导致Th1/Th2失衡。     

喘可治注射液治疗儿童支气管哮喘随机、双盲、安慰剂对照的临床和实验研究

目的研究喘可治注射液对儿童哮喘的临床疗效和对Th1/Th2平衡的调节。方法随机、双盲,安慰剂、平行对照研究68名5～14岁哮喘或哮喘合并过敏性鼻炎儿童,随机分为治疗组和对照组,分别接受喘可治或银杏注射液治疗。观察两组每日症状评分,早、晚最大呼气峰流速测定(PEF),肺功能检查(FEV1)。实验室检测细胞内细胞因子IFN-γ/IL-4的表达率。结果治疗后症状评分治疗组中位数为6.0(P25～P75为4.9～21.5),对照组为10.0(P25～P75为6.19～27.5),两组比较差异有显著性;PEF变异率治疗组为4.3%±5.1%,对照组为5.9%±6.7%,两组变异率趋势曲线差异有显著性;治疗组FEV1增加值(△FEV1)较对照组显著改善;治疗后治疗组CD4 淋巴细胞内表达IFN-γ与对照组比较有显著升高。结论喘可治治疗有助于改善哮喘患儿临床症状,降低PEF变异率,改善肺功能,并且对Th1/Th2细胞的平衡具有积极的调节作用,促进Th1细胞的优势表达,有利于哮喘症状的改善和长期稳定。     

儿童恶性骨肿瘤患者Th1/Th2亚群漂移的研究

儿童恶性骨肿瘤的发病率仅次于造血系统恶性肿瘤,其中成骨肉瘤约占儿童恶性肿瘤的5%。本病对手术及放化疗的效果不佳,预后极差[1]。随着肿瘤免疫学的发展,以细胞因子为核心的免疫治疗逐渐被临床所接受。根据分泌细胞因子的不同,可将辅助性Ｔ淋巴细胞亚群分为辅助性亚群1(Ｔｈ1)和辅助性亚群2(Ｔｈ2)两个亚群。Ｔｈ1亚群与Ｔｈ2亚群平衡状态的失调,必然导致细胞因子网络的紊乱,从而影响细胞免疫和体液免疫的功能。通过检测细胞因子的水平,可以反映Ｔｈ1/Ｔｈ2的平衡状态[2]。恶性骨肿瘤患儿体内Ｔｈ1/Ｔｈ2状态尚未见报道。本实验对此进行了…     

补充维生素D对佝偻病大鼠Th1/Th2细胞平衡的影响

目的 通过观察维生素D( VitD)缺乏性佝偻病大鼠在不同剂量VitD治疗前后免疫指标变化,探讨VitD对佝偻病大鼠Th1/Th2细胞平衡的影响.方法 选用21日龄SD大鼠32只,随机分为对照组(C组,n=10)、模型组(M组,n=10)和实验组(T组,n=12),T组又依据VitD治疗剂量不同均分为T1组(n=6)和T2组(n=6).T1组与T2组分别采用不同等级VitD一次性肌肉注射后行组间比较,观察幼鼠体质量、外周血碱性磷酸酶(AKP)、骨组织病理切片及外周血25-(OH)D3、IL-4、干扰素γ(IFN-γ)变化,并通过其外周血IL-4/IFN-γγ水平判断VitD对佝偻病大鼠Th1/Th2细胞平衡的影响.结果 缺乏VitD避光饲养30 d后,M组与T组的外观、骨组织病理切片较C组明显改变,其外周血AKP水平升高,同时血25-(OH)D3水平下降(Pa＜0.05).治疗后,T组较M组外观及病理切片有所好转,外周血AKP水平与25-(OH)D3水平明显改善(Pa＜0.05).免疫指标方面,血IFN-γ治疗前后的轻微波动均无统计学意义(Pa＞0.05),而外周血IL-4水平则在治疗后明显升高,且剂量越大,升高越明显(P＜0.05,0.01).结论 大剂量VitD可使机体Th1/Th2细胞平衡向Th2细胞移动,体内IL-4分泌增加,从而使机体处于易发生变态反应的状态.     

急性特发性血小板减少性紫癜儿童T-bet和GATA3的表达

目的：通过分析急性特发性血小板减少性紫癜（ITP）儿童血中转录因子T-bet和GATA3 mRNA的表达及细胞因子IFN-γ和IL-4的含量,探讨急性ITP发病机制中Th1/Th2细胞分化的趋势及其与转录因子的关系。方法：利用RT-PCR技术,测定急性TIP患儿发作期（n=30）和缓解期（n=28）外周血中T-bet和GATA3 mRNA 的表达,并使用ELISA法检测血浆IFN-γ和IL-4含量。20例健康儿童作为对照组。结果：ITP患儿发作期血中T-bet mRNA表达和IFN-γ含量明显高于缓解期和对照组（P＜0.01）。发作期血中GATA3 mRNA表达和IL-4的含量明显低于缓解期和对照组（P＜0.01）。结论：儿童急性ITP存在明显的Th1优势分化现象,且可能与转录因子T-bet和GATA3的调控作用有关。［中国当代儿科杂志,2010,12（1）：29-31］     

不同浓度地塞米松和甲泼尼龙对支气管哮喘患儿外周血单个核细胞免疫功能的影响

目的 观察不同浓度地塞米松(DEX)和甲泼尼龙(MP)对支气管哮喘患儿外周血单个核细胞(PBMC)中CD4+T淋巴细胞中2个功能性亚群(Th1/Th2)功能状态的影响.方法 选择2001年6月-2002年6月在重庆医科大学附属儿童医院就诊的15例哮喘患儿和14例健康体检儿童为研究对象.分为哮喘对照组、10-7mol·L-1DEX干预组、10-8mol·L-1DEX干预组、10-9mol·L-1DEX干预组、10-7mol·L-1MP干预组、10-8mol·L-1MP干预组、10-9mol·L-1MP干预组.清晨取其空腹静脉血5 mL,肝素抗凝,采用密度梯度离心法分离PBMC,加植物血凝素进行刺激培养,分别用10-7mol·L-1、10-8mol·L-1及10-9mol·L-1DEX或MP体外干预培养48 h.同期采集健康儿童空腹静脉血,同法分离培养.用ELISA法测定培养上清中γ干扰素(IFN-γ)、IL-4、IL-10及IL-12水平,并计算不同浓度DEX或MP对PBMC分泌细胞因子的抑制率.结果 1.哮喘对照组PBMC分泌IL-4水平显著高于健康对照组(P<0.05),IFN-γ/IL-4比值较健康对照组显著降低(P<0.05),2组间IFN-γ、IL-10及IL-12水平比较差异均无统计学意义(Pa>0.05).2.DEX和MP均可明显抑制哮喘患儿PBMC分泌IFN-γ、IL-4及IL-12,但对IL-10抑制作用差;与健康对照组比较,哮喘组DEX和MP抑制PBMC分泌IL-10的作用弱,差异有统计学意义(Pa<0.05).3.DEX和MP均以浓度依赖方式抑制哮喘患儿PBMC分泌IL-4,DEX在10-9mol·L-1时有促进IL-4分泌的效应,MP在10-9mol·L-1时可抑制IL-4的分泌,二组比较差异有统计学意义(P<0.05). 若以IFN-γ/IL-4表示Th1/Th2间的平衡,则MP可恢复Th1/Th2平衡(P<0.05).结论 DEX和MP均可抑制Th1/Th2类细胞因子分泌,但MP有助于恢复Th1/Th2平衡,提示临床选择MP治疗支气管哮喘更有利.     

血浆白三烯B4与儿童过敏性紫癜免疫机制的相关性

目的 探讨血浆白三烯B4(LTB4)在过敏性紫癜(HSP)患儿免疫发病机制中的作用.方法 2009年5月-2010年3月河北医科大学第二医院儿科住院HSP患儿55例(HSP组),同时取健康同龄儿童20例作为健康对照组.采用ELISA法检测各组血浆LTB4水平,流式细胞术检测各组外周血CD4+CD25+调节性T淋巴细胞及Th17细胞比例,同时观察LTB4水平与二者的相关性.应用SPSS 13.0软件进行统计学分析.结果 1.HSP患儿血浆LTB4[(63.5±20.8) ng·L-1]与健康对照组[(34.5±10.0) ng·L-1]比较明显升高(P＜0.01);2.CD4+CD25+调节性T 淋巴细胞[(3.81±1.10)%]与健康对照组[(4.45±0.90)%]比较明显降低(P＜0.05);3.Th17细胞[(1.80±0.66)%]与健康对照组[(0.52±0.24)%] 比较明显升高(P＜0.01);4.血浆LTB4水平与CD4+CD25+调节性T 淋巴细胞呈显著负相关(r=-0.67,P＜0.05),与Th17细胞呈显著正相关(r=0.57,P＜0.05).结论 LTB4可能参与HSP患儿细胞免疫功能紊乱.     

哮喘儿童细胞因子信号转导抑制因子mRNA表达与Th1/Th2的关系

目的：细胞因子信号转导抑制因子(SOCS)对JAK-STAT途径的细胞因子如白介素、干扰素等的调节起重要作用,目前SOCS与哮喘的关系仍在研究中。本研究观察SOCS1和SOCS3 mRNA在哮喘儿童外周血单个核细胞（PBMC）中的表达水平与CD4+ T细胞IFN-γ/IL-4平衡及特异性IgE（sIgE）的关系。方法：采集44例4~14岁过敏性哮喘患儿及30例健康儿童PBMC,分别用流式细胞仪分析CD4+ T细胞IFN-γ/IL-4比值,另提取总RNA,采用SYBR Green I逆转录荧光定量PCR的方法检测每组SOCS1和SOCS3 mRNA的表达。结果：哮喘组患儿外周血IFN-γ阳性的CD4+T细胞百分比［（15.7±2.0）%］及IFN-γ/IL-4比值（3.4±1.5）均低于对照组［分别为（19.1±2.7)%、4.8±2.9］;而SOCS1 mRNA（⊿Ct值11.1±1.9）表达显著高于对照组（⊿Ct值12.6±2.8）。两组儿童SOCS1 mRNA表达均与外周血分泌IFN-γ的CD4+ T细胞百分比呈负相关（P<0.05）。SOCS1和SOCS3与sIgE均无相关性。结论：SOCS1 mRNA在哮喘组患儿外周血中高表达,并与Th2占优势的免疫失衡有关。     

儿童1型糖尿病T_H1/T_H2免疫应答状况研究

目的　研究儿童 1型糖尿病TH1 TH2免疫应答状况。方法　测定 30例儿童 1型糖尿病和 30例健康儿童的干扰素γ(IFN γ)、IL 4水平。根据不同病程、不同糖化血红蛋白 (HbA1c)水平及自身抗体的存在与否进行比较。结果　 1型糖尿病患儿的IFN γ水平高于对照组 ,但差异无显著性。其中病程 <2年者 ,IFN γ水平高于病程≥ 2年者 ,但差异无显著性。 1型糖尿病患儿的IL 4水平明显低于对照组 (P <0 0 1)。IFN γ IL 4比值明显高于对照组 (P <0 0 1)。HbA1 c <7 0 %、HbA1 c 7 0 %～9 0 %及HbA1 c>9 0 %的三组患儿作IFN γ、IL 4水平及IFN γ IL 4比值比较 ,差异无显著性。自身抗体阳性患儿和自身抗体阴性患儿的IFN γ、IL 4水平及IFN γ IL 4比值比较差异无显著性。结论　儿童 1型糖尿病存在TH1细胞免疫应答增强 ,TH2细胞免疫应答减弱 ,且这种特征在糖尿病发病早期表现得更突出。     

A case of severe allergic reaction to cooked potato

White potato is a very common ingredient in the diet of infants in Mediterranean countries, and in its cooked form, it is one of the first solid foods introduced, usually around the age of 4-6 months. Allergy to potato is uncommon, and allergic reactions to cooked potato have been reported only in children. We report a case of severe potato-induced allergic reaction in an 8-month-old infant with atopic dermatitis and multiple food allergies that raises questions about differential diagnosis between anaphylaxis and food protein-induced enterocolitis syndrome (FPIES). CONCLUSION: Allergy to cooked potatoes could be a cause of severe although rare allergic reactions; it could be very difficult, in some cases, to make a differential diagnosis between anaphylaxis and FPIES. Moreover, the diagnosis appears to be very important for the choice of therapy and long-term allergologic management.     

Age-related changes in intracellular cytokine profiles and Th2 dominance in allergic children

The unbalanced T helper response has been pointed out in allergic diseases. Especially in childhood, it is important to consider the development of acquired immunity. We investigated the relationship between age and Th1, Th2, Tc1 or Tc2 cells. In addition, Th1, Th2, Tc1 or Tc2 cells in allergic diseases were compared with control subjects. Thirty-four healthy controls (0-40 years old), 200 samples of cord blood, nine patients with atopic dermatitis (AD) (1-3 years old) and five patients with bronchial asthma (BA) (2-6 years old) were studied. Surface staining with CD4, CD8 and intracellular staining with anti-interferon-gamma (IFN-gamma) and anti-interleukin (IL)-4 were carried out, and analyzed by using flow cytometry. In the healthy controls, the percentages of Th1, Tc1 or Th2 showed positive correlation with age. The absolute numbers of Th1 or Tc1 also correlated with age. Cord blood with a family history of allergic disease showed no significant difference compared to that without a family history. The percentage of Th2 in AD and BA patients was significantly higher than in the age-matched healthy controls. The increase in Th1, Th2 and Tc1 with age might reflect on the development of acquired immunity. Age matching is important when evaluating the cytokine profiles of T cells. In allergic diseases, although cord blood showed a Th1-dominant pattern, it changed to Th2 dominance in childhood, and this may reflect on some genetic background.     

Lymphocyte subsets in whole blood and isolated mononuclear leucocytes of healthy infants and children

The determination of lymphocyte subsets utilizing monoclonal antibodies and flow cytometry has become essential in the evaluation of immunological status. Using a standardized method it was found that in healthy children the percentage of CD 8+ (Leu 2+) positive cells increases significantly (P<0.01) during infancy, whereas the percentage of CD 4+ (Leu 3+) positive cells decreases with age (P<0.01). The percentage of CD 3+ (Leu 4+) cells remains constant. The ratio of CD 4/CD 8 positive cells is significantly (P<0.001) higher in infants than in older children. Other subpopulations (HLA DR+, Leu 7) were found to be constant in all age groups. For the comparison of data on lymphocyte subsets obtained by flow cytometry a standardized test procedure is important.     

特发性血小板减少性紫癜患儿Th亚群细胞因子的检测

目的探讨Th亚群细胞因子在儿童特发性血小板减少性紫癜(ITP)发病中的变化及作用。方法采用BioPlex系统和悬浮阵列技术通过荧光编码检测43例ITP患儿血清中的Th1型细胞因子TNFα、IFNγ、IL2、GMCSF和Th2相关细胞因子IL4、IL5、IL6、IL10的变化,并与20例正常儿童作对照。结果与正常健康儿童相比,ITP患儿Th1型细胞因子显著升高(P<0.01),而Th2型细胞因子显著降低(P<0.01)。结论儿童ITP发病中存在异常的淋巴细胞极化状态,是一种Th1优势的疾病。     

Early intervention with suplatast tosilate for prophylaxis of pediatric atopic asthma: A pilot study

The onset of asthma may be related to Th2 cytokine dominance at the time when food allergies occur several months after birth. This study investigated the effectiveness of early intervention with a Th2 cytokine inhibitor (suplatast tosilate) for prevention of asthma in infants with food allergies and atopic dermatitis. Suplatast tosilate dry syrup (6 mg/kg daily) or a histamine H₁-blocker (ketotifen fumarate dry syrup: 0.06 mg/kg daily) was administered randomly to 53 infants with atopic dermatitis caused by food allergies. The primary endpoints were the incidence of asthma and the time to the onset of wheezing. The peripheral blood Th1/Th2 ratio, total IgE level, and eosinophil count were measured before and after treatment. After 24 months of treatment, the prevalence of asthma was significantly lower in the suplatast group (20.8%) than in the ketotifen group (65.6%, p < 0.01). Additionally, the time from the start of treatment to the initial episode of wheezing for infants who developed asthma was significantly longer in the suplatast group than the ketotifen group (p < 0.01). Furthermore, the eosinophil count was significantly decreased by suplatast treatment (p < 0.05), and there was a significant difference between the suplatast and ketotifen groups with respect to both the eosinophil count (p < 0.01) and the Th1/Th2 ratio (p < 0.05). The results of the present pilot study suggest that suplatast tosilate is useful for the primary prevention of wheezing and asthma in children.     

1型糖尿病患儿T细胞亚群、细胞因子的变化及意义

目的　分析 1型糖尿病患儿外周血T细胞亚群 (CD3+ 、CD4 + 、CD8+ )及血清白细胞介素 2 (IL 2 )、可溶性白细胞介素 2受体 (sIL 2R)水平变化 ,以了解其在 1型糖尿病发病中的作用。方法　应用免疫荧光标记技术和流式细胞仪检测 30例 1型糖尿病患儿外周血T淋巴细胞亚群 ,同时用ELISA法定量检测患儿血清IL 2 /sIL 2R水平 ,并与正常对照组 16例作比较 ,且与胰岛功能进行相关分析。结果　 1.1型糖尿病患儿CD3+ 、CD4 + 细胞均显著升高 ,CD8+ 细胞显著降低 (P均 <0 .0 1)。 2 .1型糖尿病患儿血清IL 2明显高于正常对照组 (P<0 .0 1) ;sIL 2R较对照组明显降低 (P <0 .0 1)。IL 2与反映胰岛功能C 肽 (C P)、胰岛素呈负相关 ,而sIL 2R与C P、胰岛素呈正相关。结论　T淋巴细胞亚群失衡、IL 2及其受体参与介导胰岛 β细胞的损伤和 1型糖尿病发生     

Lymphocyte subsets and cytokines in adenoviral infection in children

To determine the distribution of major blood lymphocyte subsets we evaluated blood lymphocytes by flow cytometry in adenovirus-infected infants aged 30–730 d. In addition, interleukin-1-receptor antagonist, interleukin-10 and transforming growth factor-β1 were measured in serum by enzyme-linked immunosorbent assay. According to clinical parameters, mechanical ventilation and outcome, infections were classified as moderate ( n = 15), severe ( n = 11) and fatal ( n = 12). Controls were 13 healthy children. In severe and fatal infection, T cells (CD5⁺/CD19^-), NK effectors (CD16⁺), CD4⁺ T subset and B1 subset of B lymphocytes (CD5⁺/CD19⁺) were all significantly decreased. CD8⁺ cells were decreased in severe but not fatal cases. There was no difference in serum values of interleukin-10; however, fatal cases had high interleukin 1-receptor antagonist values. Interestingly, patients with moderate infection showed significantly increased values of transforming growth factor-β1. These results demonstrate that life-threatening adenoviral infection is associated with marked abnormalities in blood lymphocyte and cytokine profile.     

Serum measurement of thymus and activation-regulated chemokine/CCL17 in children with atopic dermatitis: elevated normal levels in infancy and age-specific analysis in atopic dermatitis

Elevated blood levels of thymus and activation‐regulated chemokine (TARC)/CCL17 have been observed in atopic dermatitis (AD) and may serve as a new biomarker for AD. However, the normal levels, especially in children, have not been well determined. We sought to establish an efficient enzyme‐linked immunosorbent assay (ELISA) with a wide range of detection that would be suitable for measurement of serum TARC/CCL17 and to determine the normal ranges of this chemokine in different age groups and its diagnostic usefulness for AD. A sensitive specific ELISA for TARC/CCL17, which we previously reported, was modified to accommodate the wide range of TARC/CCL17 values often found in sera. Twenty‐seven children with AD under 6 yr of age and 25 age‐matched normal non‐atopic controls, and 18 patients with AD and 27 controls who were 6 yr and older were enrolled. The severity of AD was evaluated using the SCORAD index. The serum levels of TARC/CCL17 were measured with the ELISA, and the serum levels of IP‐10/CXCL10 were also measured. With the novel ELISA system, the assayable range of TARC/CCL17 was 14–8000 pg/ml, and the coefficient of variation at various concentrations ranged from 2.3% to 5.0%. The serum levels of TARC/CCL17 in normal individuals were significantly higher in young children, especially in the age group of 0–1 yr. The cut‐off values of TARC/CCL17 for the diagnosis of AD were 1431 pg/ml for 0–1 yr group, 803 pg/ml for 2–5 yr group and 510 pg/ml for the 6 yr and older group, with high sensitivity and specificity of 0.83 and 0.93, 0.83 and 0.92, 0.85 and 0.96, respectively. The magnitude of the decrease in the SCORAD index after treatment with topical steroids correlated significantly with the decrease in serum TARC/CCL17. There was no difference in the serum levels of IP‐10/CXCL10 between AD and the controls. The TARC/CCL17:IP‐10/CXCL10 ratio tended to be higher in the control children aged 0–1 yr than in those aged 2–5 yr. The serum level of TARC/CCL17 reflects the severity and therapeutic response in AD. The high normal levels in infants should be taken into account when assaying TARC/CCL17.