Method of birth alters interferon-gamma and interleukin-12 production by cord blood mononuclear cells

Many uncertainties exist regarding the capability of cord blood mononuclear cells (CBMC) to produce cytokines. A number of conflicting reports led us to examine the effects of method of birth on CBMC production of interferon-gamma (IFN-γ), interleukin-4 (IL-4) and interleukin-12 (IL-12). While constitutive production of IL-4 was found in both vaginally and cesarean-delivered infants, constitutive IFN-γ or IL-12 production was found in neither. CBMC from vaginally delivered infants responded to stimulation with concanavalin A/phorbol 12-myristate 13-acetate (Con A/PMA), phytohemagglutinin (PHA), and lipopolysaccharide (LPS) with significantly higher levels of IFN-γ than CBMC from unlabored cesarean section (CS) infants. Production of IL-12 was increased in the vaginally delivered group in response to LPS and PHA but not to ConA/PMA. In contrast, mode of delivery was not associated with differences in IL-4 production. These results indicate that mode of delivery significantly alters the capability of CBMC to produce some cytokines and therefore should be taken into account in interpreting fetal/neonatal mononuclear cell function studies.     

免疫磁珠法及羟乙基淀粉法分离脐血单个核细胞的比较

目的：该实验比较了CD133免疫磁珠法及羟乙基淀粉法分离脐血单个核细胞（MNCs）的特点,探讨一种相对较好的MNCs分离方法。方法：取2007年10月至2008年3月新疆医科大学第一附属医院足月妊娠健康产妇的脐血15份,每份脐血分别用羟乙基淀粉沉淀法、CD133免疫磁珠法处理。分离后计数并在倒置显微镜下观察原代细胞的生长情况及其形态特征,原代第30天通过流式细胞仪检测CD34阳性率。结果：羟乙基淀粉沉淀法、CD133免疫磁珠组得到的MNCs数量分别为（15.23±4.30）×106/mL,（0.066±0.027）×106/mL（P＜0.05）。羟乙基淀粉沉淀法所得细胞大多悬浮生长,传代后生长缓慢;CD133免疫磁珠法的细胞生长状态良好,其CD34阳性率分别为10.1％、0.5％。结论：羟乙基淀粉沉淀法是一种高效的脐血细胞分离方法,但细胞生长状态欠佳;而 CD133免疫磁珠法所获细胞纯度较高,可根据不同要求选用相应分离法。［中国当代儿科杂志,2009,11（9）：757-760］     

Evaluation of cytotoxic function and apoptosis in interleukin (IL)-12/IL-15-treated umbilical cord or adult peripheral blood natural killer cells by a propidium-iodide based flow cytometry

Both deficient natural killer (NK) cell effector function and increased propensity to apoptosis of neonatal NK cells contribute to the increased susceptibility to infection in the neonates. Interleukin (IL)-12 and IL-15 are two immunoregulatory cytokines known to enhance cytolytic function of neonatal NK cells. The present study aims to simultaneously investigate the effect of IL-12/IL-15 on K562 cytotoxicity as well as NK cells apoptosis of enriched umbilical cord blood (CB) and adult peripheral blood (APB) NK cells, using flow cytometric cytotoxicity assays. The results indicated that (i) prior to cytotoxicity assays, CB NK cells underwent greater degree of spontaneous apoptosis than did APB NK cells; (ii) both IL-12 and IL-15 inhibited the spontaneous apoptosis of CB NK cells, while IL-15 promoted the apoptosis in APB NK cells; (iii) the deficient K562 cytotoxicity of CB NK cells could be enhanced to levels comparable with that of APB NK cells by IL-15; (iv) IL-15 increased the percentages of apoptosis in NK–K562 conjugates in a dose-dependant manner in both CB and APB with a greater effect seen with APB NK cells; (v) target-induced apoptosis was observed with APB NK cells which were further enhanced with IL-15. However, CB NK cells, unstimulated or IL-15-activated, were resistant to K562-induced apoptosis. Thus, the multi-parameter flow cytometry analysis not only demonstrates better for the deficient CB NK function but also provides greater details for cytotoxic mechanisms of NK cells.     

诱导脐血单个核细胞分化为树突状细胞的实验研究

目的：诱导脐血源树突状细胞对恶性血液病脐血移植后的过继免疫治疗十分重要。诱导条件对树突状细胞产率影响很大。本研究观察粒细胞-巨噬细胞集落刺激因子(GM-CSF)、IL-4及TNF-α分阶段诱导脐血单个核细胞来源树突状细胞的效果。方法：通过Ficoll-Hypaque法分离脐血单个核细胞,以GM-CSF、IL-4及TNF-α分阶段诱导脐血来源树突状细胞的生成,培养12 d收获细胞,以光镜和电镜观察其细胞形态学特点,以流式细胞术检测其表面标记。结果：经培养12 d,每2×10 6脐血单个核细胞收获树突状细胞数为(0.46±0.13)×106,CD80+、CD86+细胞比例分别为89.98%±4.32%、86.86%±7.17%,而CD1a+细胞比例43.16%±5.80%,经光镜和电镜下观察细胞具有典型的树突状细胞形态学特征。结论：通过GM-CSF、IL-4及TNF-α的树突状细胞培养体系和分阶段诱导法可高效率诱导脐血树突状细胞。［中国当代儿科杂志,2004, 6(5): 377-380］     

Interleukin-15 enhances CD4+ CD45RA+ expression on umbilical cord blood mononuclear cells

The reduced incidence of graft‐vs.‐host disease following umbilical cord blood (CB) transplantation may be related to the functional immaturity of newborn T cells expressing mainly the naive CD45RA phenotype. Expansion of CD4⁺ CD45RA⁺ T cells using cytokines may benefit neonates and infants with human immunodeficiency virus (HIV) infection, as a preferential decline in CD4⁺ CD45RA⁺ cells has been noted as HIV disease progresses. The aim of the study was to investigate the effect of interleukin (IL)‐15, a novel cytokine similar to IL‐2 in biological activities, on CD45RA/RO expression and apoptosis in umbilical cord blood (CB) and adult peripheral blood (APB) mononuclear cells (MNCs). Prior to culture, CB MNCs contained a greater number of CD4⁺ CD45RA⁺ cells and fewer CD4⁺ CD45RO⁺ cells than did APB MNCs. When incubated with RPMI‐1640 containing 10% fetal calf serum for 7 days, the percentage of CD45RA⁺ cells within CD4⁺ T cells (%CD45RA⁺/CD4⁺) significantly decreased compared to that of fresh CB MNCs. IL‐15 exerted a dose‐dependent increase of %CD45RA⁺/CD4⁺ and a corresponding decrease of %CD45RO⁺/CD4⁺ in CB MNCs, an effect not observed with APB MNCs treated with IL‐15. The percentages of CD45RA⁺ and CD45RO⁺ expression within CD8⁺ cells, however, were not influenced by IL‐15, in either CB or APB MNCs. A greater number of CB MNCs underwent apoptosis than did APB MNCs after 7 days of culture in RPMI‐1640 containing 10% fetal calf serum. IL‐15 did not inhibit apoptosis but induced proliferation comparable to that achieved in APB MNCs. The ability of IL‐15 to preferentially enhance the proliferation of CD4⁺ CD45RA⁺ cells in CB MNCs suggests a role for immunomodulative therapy in HIV‐infected newborns and infants.     

黄芩苷体外诱导脐血间充质干细胞向神经元样细胞分化的实验研究

目的探讨中药黄芩苷体外诱导人脐血间充质干细胞(MSCs)向神经元样细胞分化的可行性及其可能的机制。方法采集健康孕妇足月顺产儿的脐带血,共5人份,以肝素抗凝,采用明胶沉降加密度梯度离心两步法分离脐血单个核细胞,加入含黄芩苷50μmol/L的液体培养体系中进行扩增培养。取黄芩苷体外扩增2周的人脐血MSCs,采用黄芩苷诱导24h后,继续维持诱导6d,诱导30min后开始在倒置显微镜下动态观察脐血MSCs生长情况及诱导前后形态学变化。免疫细胞化学染色评价神经细胞特异性烯醇化酶(NSE),微管相关蛋白2(MAP-2)阳性细胞的表达。诱导液(DMEM培养基,200~400μmol/L黄芩苷),37℃,5%CO2诱导24h。维持诱导液(DMEM培养基,200~400μmol/L黄芩苷,B27)继续维持诱导1周。实验共分4组,分别为诱导组;对照1组(诱导液和维持液均不含黄芩苷)、对照2组(诱导液和维持液含3mmol/L的β-巯基乙醇,不含黄芩苷)、对照3组(诱导液和维持液含20g/L二甲基亚砜和20mmol/L丁化羟基苯甲醚,不含黄芩苷)、对照4组(诱导液和维持液均含有上述浓度的黄芩苷、β-巯基乙醇、二甲基亚砜和丁化羟基苯甲醚),各组分别在诱导6h、24h、7d留取标本,制作细胞爬片,细胞固定后,免疫细胞化学染色评价NSE和MAP-2阳性细胞的表达率;Hoechest33258染色,评价细胞存活率。结果诱导组诱导6h后,细胞微丝收缩,原来梭形的脐血MSCs胞体已发生收缩,细胞边缘变得不规整,出现了细的突起。7d后多数细胞成锥形,交织成网,形成较典型的神经元细胞样形态结构,免疫细胞化学染色显示黄芩苷诱导组NSE、MAP-2阳性细胞表达率以及细胞存活率分别为(76.3±9.2)%、(78.5±5.5)%、(85.3±4.8)%,显著高于对照1、2、3组(P<0.01),分别为(4.6±0.6)%、(0.7±0.6)%、(46.7±9.2)%;(63.3±6.8)%、(40.9±5.1)%、(66.5±5.2)%和(71.6±4.7)%、(42.3±4.5)%、(72.8±7.6)%。此外,各组胶质纤维酸性蛋白(GFAP)的表达率均低于1%。结论低浓度的黄芩苷体外扩增2周后的MSCs中已有少量表达NSE的阳性细胞出现,这在一定程度上黄芩苷起到了预诱导分化作用;黄芩苷能够体外诱导脐血MSCs分化为神经元样细胞,诱导过程中黄芩苷的诱导作用温和、稳定而持久;诱导出的神经元样细胞成活时间长,其诱导机制可能与黄芩苷的抗氧化、调控细胞NF-κB的活性从而刺激多种细胞因子的表达和生成有关。     

Increase of cord blood cytokine-producing T cells in intrauterine infection

BACKGROUND: Although infection is a frequent and important cause of morbidity and mortality in the neonatal period, evaluation of the immune system in cases of intrauterine infection is not easy. The subsets of T helper (Th) 1, which produce mainly interferon gamma (IFN-gamma), and Th2, which produce interleukin (IL) -4, have been implicated in the regulation of many immune responses. In this study, we investigated Th1 and Th2 subsets in the cord blood (CB) to evaluate the role of CB T cells in the intrauterine infections. METHODS: We used an intracellular cytokine-staining technique with determination by flow cytometry to study IFN-gamma-producing T cells and IL-4-producing T cells in the CB of six neonates with perinatal intrauterine infection and 17 uninfected neonates. RESULTS: The CB from neonates with intrauterine infections had more IFN-gamma-producing CD3+T cells than that from uninfected neonates. The percentage of CB IFN-gamma-producing CD3+T cells in the infected neonates correlated with the duration of membrane rupture before the onset of labor, but not with the level of C-reactive protein. The infected neonate born after the longest duration of membrane rupture showed an increased percentage of IL-4-producing CD3+T cells. CONCLUSIONS: Our results suggest that the increase of CB IFN-gamma and IL-4- producing T cells is part of the immune system directed against perinatal intrauterine infections.     

人脐血及脐带间充质干细胞的分离培养及表面标记分析

目的探讨从人脐带血及脐带分离和培养间充质干细胞(MSCs)的方法,并分析MSCs的表面标记。方法人脐带血按常规方法制备单个核细胞,利用MSCs贴壁生长的特性,经培养、换液、传代纯化MSCs;分离脐带华尔通胶(Wharton’s jelly),采用组织块贴壁法获得脐带MSCs并传代。将传代的MSCs冻存,1个月后再复苏,观察复苏后MSCs的生长情况。利用FACScan流式细胞仪检测脐带血及脐带细胞表面抗原。结果经过传代后,贴壁细胞形态趋于同一。人脐血及脐带MSCs体外生长形态相似,类似成纤维细胞,可以稳定增殖和传代。经冷冻保存,复苏后仍能较好生长。人脐血及脐带来源的MSCs表面标记CD29、CD44、CD59高表达,而表面标记CD14、CD33、CD34和CD45低表达。结论人脐带血及脐带均可分离出MSCs,在体外能扩增纯化及冻存复苏,为组织工程提供丰富的细胞来源。     

Effect of interleukin-7 and -15 on activation of purified umbilical cord blood and adult peripheral blood CD4+ T cells

We investigated the effect of two IL-2 receptor gamma chain (IL-2R-gammac)-signaling cytokines, IL-7 and IL-15, on the activation of purified CD4+ lymphocytes from umbilical cord blood (CB) and adult peripheral blood (APB) by assessing the expression of the IL-2 receptor-alpha (CD25) and Fas (CD95) on CD45RA+ (na?ve) and CD45RO+ (memory) CD4+ subsets. Induced CD40L (CD154) expression following phorbol 12-myristate 13-acetate and ionomycin (P+I) stimulation was also examined on cultured CB CD4+ cells. Incubation with IL-15 at 10 ng/ml resulted in a significant increase in the mean fluorescence intensity (MFI) of CD25 on CB CD4+/CD45RA+ cells (exceeding those of APB) without affecting the percentage of CD25-expressing CD45RA+ cells (%CD25/CD45RA). In contrast, both CD25 MFI and %CD25/CD45RA were enhanced on IL-15-treated APB CD4+ cells. CD95 expression (both percent expression and MFI) on CB CD4+/CD45RA+ cells were also enhanced by IL-15, but to a lesser extent compared to the response of ABP. IL-7, used at a concentration equivalent to IL-15, had little effect on APB CD25/CD95 and CB CD25 expression, but did enhance %CD95 expression on CB CD4+/CD45RA+ cells. Both IL-7 and IL-15 could augment the P+I-induced CD40L expression on CB CD4+/CD45RA+ T cells. However, the enhancing effect of IL-15 on CB CD40L/CD45RA expression was more sustained than that of IL-7. Thus, our study demonstrated differential activation of CB CD4+/CD45RA+ cells in response to IL-7 and IL-15 compared to adult counterparts, and the different T-enhancing function between IL-15 and IL-7.     

脐血造血干细胞移植研究进展

随着血液学和移植免疫学的快速发展,尤其是人类白细胞抗原(HLA)配型技术的不断发展,造血干细胞移植(hematopoietic stem cell transplantation,HSCT)技术逐渐成熟并广泛应用,成为治愈某些血液病的重要方法,并有望在今后的生物学治疗和基因治疗中发挥重要的作用.     

脐血间充质干细胞对T淋巴细胞增殖和激活影响的实验研究

目的研究脐血源间充质干细胞(UCB-MSCs)对T淋巴细胞激活和增殖的影响。方法2006—2007年广西壮族自治区人民医院从脐血中分离和培养间充质干细胞,流式细胞术检测其表面标志以鉴定纯度;用B r-du法测定细胞增殖率,观察UCB-MSCs对异体T淋巴细胞增殖的影响;流式细胞仪检测共培养体系中CD6 9细胞水平。结果从脐血获得的MSCs免疫表型分析显示,其不表达HLA-II抗原。对PHA刺激的脐带血T淋巴细胞,UCB-MSCs对其的增殖反应具有抑制作用,呈细胞数量剂量依赖性;UCB-MSCs与脐带血单个核细胞共培养体系中,CD 69细胞比例降低,与对照组比较差异有统计学意义(P<0.01)。结论UCB-MSCs抑制PHA刺激的T淋巴的增殖及激活,这种抑制特性表明,UCB-MSCs对免疫反应具负调节作用,可能为GVHD的防治提供一条新途径。     

紫癜性肾炎患儿外周血单个核细胞白细胞介素-13的变化

目的　探讨白细胞介素１３（ＩＬ－ １３） 在紫癜性肾炎（ＨＳＰＮ） 的变化。方法　应用逆转录－ 多聚酶链反应（ＲＴ－ＰＣＲ） 及双抗体夹心ＥＬＩＳＡ 法,分别检测了３０ 例健康儿童和２０ 例ＨＳＰＮ 患儿活动期和恢复期外周血单个核细胞（ＰＢＭＣ） 在植物血凝素（ＰＨＡ） 刺激下ＩＬ－ １３ ｍＲＮＡ和蛋白水平的变化。结果　①ＨＳＰＮ 活动期ＰＢＭＣ中ＩＬ－１３ ｍＲＮＡ 和蛋白水平均显著高于正常对照组（０ ．４８ ±０ ．１１ ｖｓ０．３６±０．１６ 和４７ ．８９ ±１１ ．６７ ｐｇ／ｍｌｖｓ３５．２２±４．４２ ｐｇ／ｍｌ, Ｐ均＜ ０．０１） ,恢复期与正常对照组相比无显著差异（０．３８±０．１２ ｖｓ０．３６±０ ．１６ 和３５ ．９４ ±５．６０ ｐｇ／ｍｌｖｓ ３５ ．２２±４ ．４２ ｐｇ／ｍｌ,Ｐ均＞ ０．０５）;②临床上表现为蛋白尿型、肾病综合征型及急性肾炎综合征型者,活动期ＰＢＭＣ中ＩＬ－ １３ ｍＲＮＡ和蛋白水平均显著高于正常对照组,而表现为单纯血尿型者与正常对照组相比无显著差异。结论　ＨＳＰＮ 患儿活动期ＩＬ－ １３ 表达异常,且与临床类型有一定的相关关系。     

脐血造血细胞含量与白血病脐血移植疗效分析

目的分析4711份库存脐血造血细胞含量及探讨脐血造血细胞含量与白血病脐血移植疗效的关系。方法分析4711例库存脐血总有核细胞数(TNC)和CD34+细胞数分布情况,探讨不同的造血细胞输入量、供受者HLA不相合数、受者性别、年龄、体重和疾病类型间植入率和生存率的差异。结果 4711例库存脐血TNC和CD34+细胞中位数分别为1.14×109/kg和4.06×106/kg,按3.7×107/kg有效TNC输入量计算,93.2%脐血可供体重50 kg以下受者移植。89例白血病患者移植后植入75例,植入率为84.3%。中性粒细胞绝对值≥0.5×109/L、血小板≥20×109/L和≥50×109/L的时间分别为移植后17、34和46 d。75例植入病例中,长期无病存活47例,死亡26例,2例复发;急性移植物抗宿主病(GVHD)Ⅰ～Ⅱ度、Ⅲ～Ⅳ度和慢性GVHD发生率分别为54.7%、20.0%、9.3%。影响移植植入率的因素包括受者年龄、TNC和CD34+细胞输入量;影响生存率的因素包括受者年龄、体重和输入CD34+细胞数。结论在无法找到HLA全相合骨髓供者时,可选择脐血作为替代骨髓的造血干细胞来源治疗儿童与成人白血病,TNC和CD34+细胞数仍是选择脐血移植物的参考指标。     

The effect of interleukin-12 and interleukin-15 on CD69 expression of T-lymphocytes and natural killer cells from umbilical cord blood

We investigated the effect of two immunoregulatory cytokines interleukin-12 (IL-12) and IL-15, alone or in combination, on CD69 expression of mononuclear cells (MNCs) obtained from umbilical cord blood (CB) and adult peripheral blood (APB). We established that (1) CD3/69, but not CD16/69, expression on CB MNCs could be increased with IL-12, IL-15 or both in 18-hour cultures, but to a lesser degree compared to that on corresponding APB MNCs, (2) CD3/69 expression on CB MNCs was significantly increased after 1 week's culture with IL-12, especially with IL-15, exceeding that on APB MNCs similarly activated and (3) CD16/69 expression on CB MNCs, but not APB MNCs, was greatly increased after 1 week's culture with IL-15. The combination of IL-12 + IL-15 resulted in greater CB CD3/69 expression than individual cytokines, while producing less of an effect on CD16/CD69 expression as compared to IL-15 alone. The results of our study indicate that neonatal T and NK cells readily respond to cytokine stimulation by upregulating CD69 expression, with a greater effect achieved using IL-15 compared to IL-12.     

Effects of cytokines on hematopoietic progenitor cells in cord blood,in bone marrow,and in peripheral blood mobilized by chemotherapy and G-CSF

We compared the effects of various combinations of cytokines (stem cell factor [SCF], interleukin [IL] ?3, IL-6, granulocyte-colony stimulating factor [G-CSF], erythropoietin [EPO]) among the growth of human hematopoietic progenitor cells from cord blood (CB), bone marrow (BM), and peripheral blood mononuclear cells (MNC) mobilized by chemotherapy and G-CSF (PB) in a semi-solid medium. Macroscopic colonies, that were visible to the naked eye, were formed from PB-MNC within 1 week even without cytokines. They consisted of blasts containing macrophage-like cells with immature nuclei on Wright stain, and were strongly accelerated by IL-3. Macroscopic colonies were also formed from CB-MNC. However, they appeared after 1–3 weeks and synergistic effects of SCF with other cytokines, especially EPO, were prominent. Macroscopic colonies were not formed from BM-MNC. Granulocyte-colony stimulating factor was effective in increasing colony forming units of granulocyte macrophage from BM-MNC and they appeared between 1 and 2 weeks. These results suggested that the quality of hematopoietic progenitor cells was different among blood sources. This might lead to different bone marrow recovery patterns after transplantation of each blood source. The appropriate cytokines should be added to evaluate their exact potential.     

Cardiac troponin I, cardiac troponin T and creatine kinase MB concentrations in umbilical cord blood of healthy term neonates

AIMS: To measure and compare cardiac troponin I, cardiac troponin T and creatine kinase MB concentrations in the umbilical cord blood of healthy term infants and to investigate the relationship between maternal and neonatal troponin values at birth. METHODS: Troponin I, troponin T and creatine kinase MB concentrations were measured from the umbilical cord samples of 85 healthy term neonates and in the blood samples of their respective mothers at birth. RESULTS: Median (interquartile range) umbilical cord concentrations were 0 microg/L (0-0) for troponin I, 0 microg/L (0-0.019) for troponin T and 4.90 microg/L (3.90-6.61) for creatine kinase MB. Troponin I and T concentrations were higher than the detection limit for the assay in 2 (2.3%) and 41 (48.2%) neonates, respectively. Two mothers (2.3%) had cTnT levels above the detection limit; none of them had increased levels of cTnI. CONCLUSION: Probably owing to differences in expression and assay detection limits, cord blood troponin T concentrations are frequently over the detection limit at birth, while troponin I is mostly undetectable and comparable with that in healthy pregnant women. These cardiac regulatory proteins are of neonatal origin and are not influenced by maternal levels.     

Similar outcomes of allogeneic hematopoietic cell transplantation from unrelated donor and umbilical cord blood vs. sibling donor for pediatric acute myeloid leukemia: Multicenter experience in China

In a multicenter study, we have conducted a retrospective study on 73 pediatric AML patients who were primary refractory or in greater than CR1 and investigated MSD (or MMSD) (n = 20), URD (n = 23), and UCB (n = 30) HCT between January 1998 and October 2009. The median day to neutrophil engraftment was similar in all groups. The median day to platelet engraftment was longer in the UCB group. The number of HLA mismatch was higher in the UCB group (p = 0.034); however, the cumulative incidence of grade III–IV aGVHD was not different among all groups (p = 0.125); furthermore, cGVHD was lower in the UCB group (p = 0.078). The risk of relapse did not differ among all groups (RR = 1.28, p = 0.125), but the patients of MSD (or MMSD) grafts had a trend of higher risk recurrence. Sixty‐two patients survived with a median follow‐up of 58.2 months. Five‐yr LFS was 73.1%, 59.8%, and 59.6% for URD, UCB, and MSD (or MMDS), respectively (p = 0.426). Five‐yr LFS in CR1 was 68.9%, with a significantly better result compared to 41.7% in CR2 (p = 0.025). Our comparisons suggest that pediatric AML patients receiving UCB had a higher early TRM, a lower cGVHD rate, and a similar long‐term survival. The outcome of URD and UCB is comparable to that of a suitable sibling for pediatric AML.     

再生障碍性贫血患儿外周血Tre g／Th 17细胞失衡的研究

目的：探讨CD4＋CD25＋调节性T细胞（Treg）/Th17细胞失衡在儿童再生障碍性贫血（AA）发病中的意义。方法采用流式细胞仪（FCM）检测AA患儿和健康对照组儿童外周血Treg细胞和Th17细胞的比例,酶联免疫吸附法（ELISA）检测血浆中IL-17和IL-6的水平。结果 AA患儿外周血Th17细胞比例（1.45±0.28）％显著高于同龄对照组（0.45±0.10）％（P＜0.01）,而Treg细胞比例（4.05±1.07）％及 Treg/Th17细胞比值（2.89±0.88）明显低于同龄对照组儿童（6.96±0.79）％、（15.77±2.77）（P＜0.01）。AA患儿血浆中IL-17、IL-6水平分别为（185.96±40.42）、（20.78±5.49）pg/mL,显著高于同龄对照组（120.47±18.39）、（10.44±2.51）pg/mL （P＜0.01）,且IL-17和IL-6水平均与Th17细胞比例呈正相关（r＝0.67,P＜0.01;r＝0.57,P＜0.01）,而Treg细胞比例与IL-6水平呈负相关（r＝-0.39,P＜0.05）。结论 AA患儿Treg细胞比例的减低和Th17细胞比例的增加所致的Treg/Th17细胞失衡,可能在儿童再障发病中起重要作用,IL-6高表达可能是导致Tre g/Th 17细胞失衡的原因之一。