儿童重症化脓性脑膜炎CD3＋ CD8＋ T细胞与炎症指标、体液免疫的变化

目的：通过观察儿童重症化脓性脑膜炎早期血CD3＋CD8＋T细胞的变化,以及与炎症指标、体液免疫指标之间的关系,探讨其在儿童重症化脓性脑膜炎发生发展中的临床意义。方法回顾性分析中国医科大学附属盛京医院PICU 2014年8月1日至2015年12月31日收治的39例1个月~14岁的重症化脓性脑膜炎患儿,血CD3＋CD8＋T细胞计数正常或升高(≥190个/mm3)为A组( n＝22),降低(<190个/mm3)为B组(n＝17),分析患儿的一般资料、血液炎症指标、体液免疫、脑脊液改变在两组患儿中的分布和差异。结果17例(43.6％)患儿CD3＋CD8＋T细胞明显下降;所有4例死亡均为B组患儿;虽然没有统计学差异,但 B 组 Glasgow 昏迷评分<8分者比例(58.8％)高于 A 组(31.8％)。B组C-反应蛋白、降钙素原中位数(最小值-最大值)分别为251.0(26.2-417.0)mg/L、32.7(0.9-100.0)ng/L,远远高于A组的106.5(12.0-458.0)mg/L、4.5(0.1-200.0)ng/L,差异有统计学意义(P<0.05);B组中6例(35.3％)外周血WBC<4×109/L,而 A组为1例(4.6％),中性粒细胞>80％的比例A组为7例(31.8％),而B组为12例(70.6％),两组比较差异有统计学意义(P<0.05)。 B组14例(82.3％)患儿脑脊液中糖含量<2.0 mmol/L,高于A组[11例(50.0％)],两组比较差异有统计学意义(P<0.05)。结论儿童重症化脓性脑膜炎CD3＋CD8＋T细胞可能受到抑制,其与患儿脑功能损伤程度、炎症反应以及预后相关。可能对指导临床免疫制剂的应用有一定帮助。     

Munc13‐4 deficiency with CD5 downregulation on activated CD8+ T cells

Familial hemophagocytic lymphohistiocytosis (FHL) is characterized by uncontrolled activation of T cells and macrophages and hypercytokinemia. We have recently described a significant increase in a subpopulation of CD8⁺ T cells with downregulation of CD5 during the acute phase of FHL type2 (FHL2; perforin deficiency), which declines after successful treatment, with a concomitant reduction in serum cytokine level. This unusual subset of CD8⁺ T cells, however, has not been characterized in patients with other subtypes of FHL. Herein, we describe a patient with FHL3 (Munc13‐4 deficiency) carrying compound heterozygous mutations in the UNC13D gene. He had high serum levels of pro‐inflammatory cytokines and significantly increased activated CD8⁺ T cells with downregulation of CD5 during the acute phase, similar to that found in FHL2. This immunophenotypic feature may serve as a useful marker of immune dysregulation in FHL3 in addition to FHL2.     

CD4~+CD25~+Treg在儿童急性免疫性血小板减少性紫癜发病机制中的作用

目的 研究CD4~+CD25~+调节性T细胞(CD4~+CD25~+Treg)在儿童急性免疫性血小板减少性紫癜(AITP)发病机制中的作用.方法 以25例AITP患儿的外周血为标本,30例健康儿童为正常对照.采用三色单克隆抗体直接标记法检测细胞膜表面抗原;检测胞浆/核抗原时先标记膜表面抗原,固定破膜后再标记胞浆/核抗原;应用多参数流式细胞仪进行Treg细胞的数值检测和结果分析;酶联免疫吸附法检测患儿和正常儿童血清中TGF-β和IL-10水平.结果 约60%的CD4~+CD25~+细胞为FoxP3阳性,而90%以上的CD4~+CD25~(hi)表达FoxP3.AITP患儿与健康儿童CD4~+CD25~(hi)、CD4~+CD25~+FoxP3~+细胞数和CD4~+CD25~+FoxP3~+与CD4~+CD25~+FoxP3~-的比值分别为(0.95±0.27)%对(1.08±0.37)%、(4.54±1.31)%对(5.28±1.52)%和(0.61±0.26)%对(0.73±0.29)%,两组间差异均无统计学意义(P均>0.05);AITP患儿和正常儿童血清的TGF-β水平分别为(9.44±2.78)ng/ml对(25.23±3.42)ng/ml,两者差异有统计学意义(P<0.05);两组间IL-10水平差异无统计学意义.AITP患儿的成熟B淋巴细胞显著高于正常儿童[(19.90±7.42)%对(12.02±3.82)%],P<0.05;NK细胞、CD3~+T细胞和CD3~+CD4~+/CD3~+CD8~+数值在两组间差异无统计学意义(P均>0.05).结论 数量正常但TGF-β分泌不足的Treg细胞,可能在儿童AITP发病中发挥一定的作用.     

CD4+CD25+调节性T细胞在支气管哮喘中的研究进展

近年来,关于CD4^＋CD25^＋调节性T细胞在支气管哮喘发病中作用的研究有相当大的进展,支气管哮喘的发病机制已不能单纯用Th1/Th2平衡理论来解释,CD4^＋CD25^＋调节性T细胞越来越受到重视,它通过细胞接触和分泌细胞因子发挥免疫抑制功能,维持自身免疫平衡,控制支气管哮喘的发展,Foxp3、IL-6等在其功能调控中发挥重要作用。支气管哮喘中,有关CD4^＋CD25^＋调节性T细胞作用机制及数量变化的观点不一,糖皮质激素通过增强CD4^＋CD25^＋调节性T细胞功能发挥治疗作用。     

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目的探讨支气管哮喘患儿外周血中辅助T细胞(Th)17细胞和CD4+CD25+调节性T细胞(Treg)的变化与儿童哮喘病情的相关性。方法收集2009年月5月至2010年4月于郑州大学第一附属医院就诊的患儿,均为首次确诊哮喘或规范吸入激素停用>3个月后复发及近1个月内无明显感染者。采用流式细胞仪测定患儿外周血中Th17细胞及CD4+CD25+Treg比例的变化。结果 Th17细胞在哮喘急性期组(2.24%±1.02%)较哮喘缓解期组(1.65%±0.38%)及健康儿童组(1.02%±0.28%)均显著增高(P<0.05),哮喘缓解期组(1.65%±0.38%)和健康儿童组(1.02%±0.28%)无明显差别,CD4+CD25+Treg细胞比例在3组儿童间差异均有统计学意义(F=45.604,P<0.05),与健康儿童组(7.11%±0.89%)相比,哮喘缓解期组(6.05%±0.87%)和哮喘急性期组(5.37%±0.80%)的比例明显下降,而哮喘急性期组较健康儿童组下降。哮喘急性期组轻、中、重度3组之间差异同样有统计学意义。Th17细胞与哮喘患儿病情呈正相关(r=0.649,P<0.05),而CD4+CD25+...     

急性白血病患儿CD4~+CD25~+调节性T细胞和NK细胞免疫作用探讨

目的观察外周血CD4+CD25+调节性T细胞(CD4+CD25+Treg)及自然杀伤细胞(na-ture killer cell,NK)在白血病患儿及非白血病患儿的不同,了解白血病患儿的免疫状态,探讨CD4+CD25+Treg细胞及NK细胞在小儿急性白血病肿瘤免疫中的意义。方法以流式细胞术检测急性白血病初诊患儿及非白血病患儿各30例的外周血CD4+CD25+Treg细胞、NK细胞的数量及比例。结果外周血CD4+CD25+CD127-细胞占CD4+T细胞的比例白血病组为(11.45±1.41)%,显著高于对照组为(6.98±1.09)%(P<0.05)。而NK细胞数量白血病组为(5.13±2.97)%,显著低于对照组为(15.06±3.91)%(P<0.05)。结论 (1)急性白血病患儿外周血CD4+CD25+Treg细胞数量升高,NK细胞数量降低,表明急性白血病患儿NK细胞免疫功能处于抑制状态。CD4+CD25+Treg细胞可能在白血病的发生、发展中起一定作用。(2)通过检测CD4+CD25+CD127-T细胞可较好的反映CD4+CD25+Treg细胞的比例,简便可行、重复性好、检测结果准确、可靠。     

Regulation of CD31 expression and interleukin-4 production by human cord blood CD4+ T cells with interleukin-4 and interleukin-7

BACKGROUND: T helper 2 cell type cytokines, such as interleukin (IL)-4 and IL-5, play pivotal roles in the development of allergic diseases. However, the mechanism by which naive CD4+ T cells acquire the ability to produce these cytokines remains unclear. Recently, it was reported that IL-7 induces the ability to produce IL-4 as well as interferon (IFN)-gamma and IL-5 in naive CD4+ T cells without TCR stimulation. To further analyze the mechanism of acquiring IL-4-producing ability by naive CD4+ T cells, the effects of IL-7 on human cord blood CD4+ T cells were compared with those of IL-4, which induced the ability to produce IFN-gamma but not IL-4. RESULTS: Interleukin-7 preserved the population of CD4+CD31- T cells in cord blood and induced their IL-4-producing ability without T cell receptor (TCR) stimulation, while IL-4 induced CD31 on CD31- T cells and could not induce their IL-4-producing ability. Both the CD31-inducing effect and the inhibitory priming effect for IL-4-production by IL-4 were also observed after cord blood CD4+ T cells had been primed with IL-7 and acquired the IL-4-producing ability. CONCLUSIONS: Interleukin-7 induced the IL-4-producing ability in naive CD4+CD31- T cells without TCR stimulation, suggesting that the signal transduction via CD31 may have an inhibitory effect on the acquisition of the IL-4-producing ability by cord blood CD4+ T cells in the absence of TCR stimulation.     

诱导性调节性T淋巴细胞在支气管哮喘患儿粉尘螨滴剂舌下特异性免疫治疗长期疗效中的作用和意义

目的 探讨诱导性调节性T淋巴细胞在粉尘螨滴剂舌下特异性免疫治疗(SLIT)长期疗效中的作用和意义.方法 选择对粉尘螨过敏的轻度支气管哮喘患儿60例,分为非SLIT组和SLIT组.非SLIT组仅接受药物治疗,而SLIT组在药物治疗基础上予舌下含服“粉尘螨滴剂”治疗,分别在治疗前、治疗1年、治疗结束时及治疗结束后1年时分离2组患儿外周血单个核细胞,予粉尘螨浸出液刺激后检测诱导性调节性T淋巴细胞水平变化;同时留取血清检测粉尘螨特异性IgE水平变化.结果 SLIT组患儿诱导性调节性T淋巴细胞水平较非SLIT组增高,并随着SLIT的进程逐渐增高,SLIT结束时达高峰并于SLIT结束1年后仍保持在较高水平;非SLIT组患儿在相应时间点诱导性调节性T淋巴细胞水平的差异无统计学意义(P＞0.05);2组患儿治疗前后特异性IgE水平差异均无统计学意义(P均＞0.05).结论 SLIT有可能通过上调诱导性调节性T淋巴细胞的功能来诱导免疫耐受;诱导性调节性T淋巴细胞在维持SLIT的长期疗效中可能发挥重要作用.     

轮状病毒感染CD4+T细胞相关细胞因子研究进展

轮状病毒是世界范围内引起婴幼儿腹泻的主要病原,除胃肠道症状外,还可引起全身多个系统病变.随着对轮状病毒感染免疫研究的不断深入,细胞免疫在其发病中的作用和机制越来越受到重视.T细胞参与了轮状病毒感染的免疫及保护,CD4+T细胞及其所分泌的相关细胞因子对轮状病毒的预防及清除起重要作用,也为疫苗的研制和肠道外感染的防治提供理...     

重症化脓性脑膜炎儿童血CD4~+CD25~+调节性T细胞的变化

目的初步探讨CD4~+CD25~+调节性T细胞(Tregs)在儿童重症化脓性脑膜炎病程早期的变化及其可能的作用。方法回顾性研究2014年8月至2015年12月入住PICU的39例严重化脓性脑膜炎患儿的临床资料,并根据患儿入院后12h内血Tregs降低与否分为无Tregs降低组和Tregs降低组(Tregs计数410个/mm3为降低),分析早期Tregs改变与临床表现、实验室指标和转归的关系。结果 39例患儿中,13例(33%)Tregs比例明显下降(31%),18例(46%)Tregs绝对计数下降(410个/mm~3)。4例病死患儿均来自于Tregs降低组。与无Tregs降低组比较,Tregs降低组患儿外周血白细胞低于正常的比例更高,血清降钙素原升高更显著(P0.05)。结论儿童重症化脓性脑膜炎患儿早期Tregs可能受到抑制,这与患儿炎症反应更重、病死率更高有关。     

哮喘患儿外周血中CD4~+LAP~+调节性T细胞比例及功能变化研究

目的测定哮喘患儿外周血中CD4+LAP+调节性T细胞(CD4+LAP+Treg细胞)比例和功能的改变,探讨CD4+LAP+Treg细胞在哮喘发病中的作用。方法选取2014年3月至2015年9月确诊为哮喘的患儿75例为研究对象,依据病情将哮喘患儿分为哮喘急性期组(n=40)和哮喘缓解期组(n=35);另选取行健康体检儿童30例为健康对照组。采用流式细胞仪检测各组儿童外周血CD4+LAP+Treg细胞百分比;采用3H-脱氧胸腺嘧啶苷法检测各组CD4+LAP+Treg细胞的免疫抑制情况。结果哮喘急性期组患儿体内CD4+LAP+Treg比例(2.0%±1.0%)较缓解期组(4.1%±2.4%)及健康对照组(4.6%±3.0%)均降低(P0.05);哮喘急性期组患儿体内CD4+LAP+Treg细胞免疫抑制率(21%±4%)较缓解期组(55%±9%)及健康对照组(62%±11%)均降低(P0.05)。结论哮喘患儿外周血中CD4+LAP+Treg数量降低和抑制功能减弱可能参与了哮喘的急性发病过程。     

肾母细胞瘤患儿外周血调节性T细胞和自然杀伤T细胞的表达研究

目的 研究肾母细胞瘤患儿外周血CD4+CD25+CD127low 调节性T细胞（Treg）和CD3+CD16+CD56+自然杀伤T细胞（NKT）的表达变化,初步探讨发生改变的原因及其临床意义。方法 选取2015年11月至2016年7月就诊的21例肾母细胞瘤患儿作为病例组,21例于本院体检的同年龄段健康儿童为健康对照组,采用流式细胞术检测两组儿童外周血中 CD4+CD25+CD127low T细胞占CD4+T细胞百分比和CD3+CD16+CD56+T细胞占CD3+T细胞百分比,分别代表Treg水平和NKT水平。结果 病例组肾母细胞瘤患儿外周血Treg水平低于健康对照组（P < 0.05）;病例组肾母细胞瘤患儿外周血NKT水平高于健康对照组（P < 0.05）。结论 Treg和NKT水平变化与肾母细胞瘤的发生与发展有关,Treg和NKT水平可能是反映肾母细胞瘤患儿免疫功能状态的较好指标。     

毛细支气管炎CD4+CD25+调节性T细胞和Foxp3 mRNA表达及与IgE关系

目的：通过对呼吸道合胞病毒(RSV)毛细支气管炎患儿CD4+CD25+调节性T细胞（CD4+CD25+Treg）、Foxp3 mRNA的表达、IgE合成的检测,以探讨其在RSV毛细支气管炎发病机制中的作用。方法：在RSV检测阳性的57例毛细支气管炎患儿中,运用流式细胞仪检测外周血CD4+CD25+Treg的比例,RT-PCR检测Foxp3 mRNA的表达量、ELISA法检测血清总IgE含量。25例健康儿童作为对照组。结果：毛细支气管炎患儿外周血CD4+CD25+Treg细胞数量减少,特应性体质（7.7±1.6%）和非特应性体质组（8.8±2.1%）均低于对照组（10.5±1.6%）（P均<0.01）。毛细支气管炎特应性体质和非特应性体质组外周血Foxp3 mRNA表达亦均低于对照组（P<0.01）。而血清总IgE含量毛细支气管炎特应性体质（241.2±102.5 IU/mL）和非特应性体质组(125.5±63.2 IU/mL)均高于对照组(27.2±10.5 IU/mL)（P<0.01）。外周血CD4+CD25+Treg细胞数量、Foxp3 mRNA表达及血清总IgE含量在毛细支气管炎特应性体质和非特应性体质组之间差异有显著性意义。CD4+CD25+Treg、Foxp3 mRNA与血清IgE水平之间相关密切,r=-0.70,-0.79(均P<0.01)。结论：毛细支气管炎外周血淋巴细胞CD4+CD25+Treg和Foxp3 mRNA表达降低,二者低水平表达使IgE合成增多,共同参与了RSV毛细支气管炎的发病。［中国当代儿科杂志,2009,11（5）：349-353］     

Reduced expression of CD69 and adhesion molecules of T lymphocytes in asthmatic children receiving immunotherapy

T lymphocytes play a fundamental role in the initiation and regulation of chronic inflammatory responses in patients with asthma. CD69 is an early marker of T‐cell activation. The levels of intercellular adhesion molecule‐1 (ICAM‐1, CD54) and L ‐selectin have been reported to increase in patients with allergic diseases and asthma. The present study was therefore undertaken to investigate the expression of CD69, CD54, and L ‐selectin by T lymphocytes of children with asthma, before and after immunotherapy. Eighteen children newly diagnosed with asthma, 11 good and nine poor responders to immunotherapy, and 16 normal subjects, were enrolled in this study. The percentages of CD69⁺, CD54⁺, and CD62L⁺ cells in T lymphocytes were measured by using flow cytometry. The levels of CD69, CD54, and CD62L in serum and culture supernatants were determined by using enzyme‐linked immunosorbent assay (ELISA). The expression of CD69 and CD54 on CD3⁺ T lymphocytes was significantly higher in children with asthma than in control patients. All the patient groups expressed (spontaneously and following stimulation with phorbol myristate acetate and ionomycin together with mite‐extract proteins) greater amounts of CD69 and CD54 than did control subjects. With long‐term immunotherapy, the percentages of CD69⁺ and CD54⁺ T lymphocytes were significantly lower in patients with a good response to immunotherapy. Our results also showed significantly lower serum L ‐selectin levels following immunotherapy. In conclusion, successful immunotherapy resulted in decreased expression and production of CD69 and CD54. These results may explain, in part, the clinical efficacy of immunotherapy.     

Th17细胞与Treg细胞在支气管哮喘发病机制中的研究进展

近年来,众多研究发现支气管哮喘的发病机制已不能单纯用Th1/Th2平衡理论来解释,CD4+ CD25+调节性T细胞和Th17细胞及其细胞因子IL-10、IL-17、转化生长因子-β等与支气管哮喘发病明显相关.由于Th17细胞与CD4+ CD25+调节性T细胞在功能上相互拮抗,而在分化上密切相关,因此这两种细胞的免疫失衡也是支气管哮喘发病的重要原因.糖皮质激素可通过维甲酸相关孤核受体γt信号途径降低IL-17的表达,还可以通过诱导转录因子Foxp3的表达调控CD4+ CD25+调节性T细胞的分化和功能.     

Responses of CD4(+) CD25(+) Foxp3(+) and IL-10-secreting type I T regulatory cells to cluster-specific immunotherapy for allergic rhinitis in children

We investigated the effects of cluster specific immunotherapy (SIT) with Dermatophagoides pteronyssinus (Der p) on CD4(+) CD25(+) Foxp3(+) Treg cells and IL-10-secreting type I T regulatory (Tr1) cells in Der p-sensitized children with allergic rhinitis (AR). We performed a prospective randomized study involving 46 children (aged 8-13 yr), of whom 25 children received Der p-SIT + pharmacotherapy and 21 received only pharmacotherapy, over a period of 1 yr. Prior to and at end of treatment, CD4(+) CD25(+) Foxp3(+) Treg cells and allergen-specific IL-10(+) IL-4(-) , IFN-γ(+) IL-4(-) , and IL-4(+) IFN-γ-CD4(+) T cells were measured by flow cytometry. Similarly, IL-4, IFN-γ, and IL-10 in supernatants from allergen-stimulated peripheral blood mononuclear cell (PBMC) cultures were measured by ELISA, and the suppressive effect of CD4(+) CD25(high) T cells on cell proliferation and cytokine release was estimated from both groups. Allergen-specific serum IgE and IgG4 were also assessed at the beginning and end of treatment by RAST and ELISA, respectively. The levels of allergen-specific Tr1 cells, IgG4, and allergen-induced IL-10 synthesis from PBMC cultures were significantly increased after SIT for 1 yr compared with baseline levels (p < 0.001 for all), with significant correlation between increased levels of Tr1 cells and improvements in nasal symptoms (r = 0.48, p < 0.05). In contrast, the levels of CD4(+) CD25(+) Foxp3(+) T cells, allergen-specific Th1 and Th2 cells, the production of IL-4 and IFN-γ, and the function of CD4(+) CD25(high) T cells were not altered in either group at the end of treatment. These data suggest that the up-regulation of Tr1 cells may play an important role in SIT and be a useful marker of successful SIT in AR patients.     

孟鲁司特钠对哮喘小鼠气道重塑及Th17/CD4~+CD25~+Treg表达的影响

目的探讨白三烯受体拮抗剂孟鲁司特钠干预哮喘小鼠后气道重塑及Th17细胞/CD4~+CD25~+调节性T细胞(CD4~+CD25~+Treg)表达的动态变化及其相关性。方法将Balb/c小鼠随机分成空白组、哮喘组、孟鲁司特钠组,每组经腹腔注射鸡卵清蛋白(OVA)和氢氧化铝混悬液致敏并雾化吸入2.5%OVA以制备哮喘气道重塑模型,空白组以生理盐水替代;孟鲁司特钠组雾化前给予孟鲁司特钠混悬液灌胃,空白组及哮喘组以生理盐水代替。3组分别在雾化2周、4周及8周后的24 h内随机处死8只小鼠。肺组织病理切片观察气道重塑程度;流式细胞技术检测脾组织中Th17、CD4~+CD25~+Treg细胞占CD4~+T细胞百分比。结果各时间点哮喘组和孟鲁司特钠组支气管总管壁厚度、平滑肌厚度均高于空白组(P0.05),干预8周时孟鲁司特钠组较哮喘组上述变化明显减轻(P0.05)。与空白组比较,各时间点哮喘组和孟鲁司特钠组均显示Th17细胞数增加,与气道重塑呈正相关(P0.05);而CD4~+CD25~+Treg细胞数逐渐降低,与气道重塑呈负相关(P0.05)。干预8周时孟鲁司特钠组与哮喘组比较,Th17细胞数显著下降(P0.05);而CD4~+CD25~+Treg细胞数明显增加(P0.05)。结论孟鲁司特钠干预哮喘小鼠后能减轻气道重塑的发生,且随着用药时间延长,改善越明显;机制可能是通过改善哮喘小鼠体内Th17/CD4~+CD25~+Treg的免疫紊乱,抑制气道炎症反应从而减轻或延缓气道重塑来起作用的。     

CD4+CD25high+CD127low调节性T 细胞在毛细支气管炎患儿外周血中的表达改变及其临床意义

目的：探讨CD4+CD25high+CD127low调节性T 细胞（regulatory T cell, Treg）在毛细支气管炎患儿外周血中的数量变化及其临床意义。方法：随机选取31例2岁以下毛细支气管炎患儿作为毛细支气管炎组,同期住院的同年龄段支气管肺炎患儿25例和患有疝气、肾结石等非感染性疾病患儿25例分别作为支气管肺炎组和对照组,采用流式细胞术检测3组患儿外周血中CD4+CD25high+CD127lowTreg 占CD4+T 细胞百分比。结果：毛细支气管炎组患儿外周血中CD4+CD25high+CD127lowTreg 占总CD4+T 细胞的8.0%±2.1%,低于支气管肺炎组（9.6%±2.6%）及对照组（11.3%±2.9%）,差异有统计学意义（P<0.05）。结论：毛细支气管炎患儿外周血CD4+CD25high+CD127lowTreg水平可能是一个较好的反映毛细支气管炎患儿免疫功能状态的参考指标,其在外周血中数量下降提示Treg细胞可能参与毛细支气管炎的发生发展。