Levels of Th1 and Th2 cytokines in children with post-infectious bronchiolitis obliterans

BACKGROUND: Post-infectious bronchiolitis obliterans (BO) is a chronic obstructive airway disease associated with inflammation and fibrosis of the small airways; it is more common in children who have had acute viral bronchiolitis. No previous studies have reported the immune response of BO. Unbalanced Th1/Th2 immune response might be one of the risk factors for developing this illness. AIM: To compare the production of interferon (IFN)-gamma, interleukin (IL)-4 and IL-10 in peripheral blood mononuclear cell cultures in children with BO and in healthy children. METHODS: From March 2003 to October 2003, children with BO and healthy children were selected from the paediatric outpatient clinics in our centre. Peripheral blood was collected and mononuclear cells were separated and cultured (96 hours) with 1% phytohaemagglutinin stimulation. The supernatant was stored and cytokine levels were measured through ELISA. RESULTS: IFN-gamma, IL-4 and IL-10 levels were not significantly different between the groups studied. Family history of atopy was significantly associated with subjects with BO (p=0.02). CONCLUSIONS: Our results suggest that unbalanced peripheral blood Th1/Th2 immune response of children with post-infectious BO might not be associated with its pathophysiology. Further studies are required to better understand the role of risk factors, including viral genotype, viral load or tissue repair abnormalities in the development of post-infectious BO.     

头孢呋辛对哮喘儿童外周血单个核细胞Th1/Th2平衡的影响

目的研究头孢呋辛对哮喘儿童外周血单个核细胞Th1/Th2平衡的影响。方法采用流式细胞术检测哮喘和健康儿童外周血单个核细胞IFN-γ和IL-4水平,以及哮喘儿童外周血单个核细胞经头孢呋辛体外干预后的IFN-γ和IL-4水平。结果与健康儿童相比,哮喘患儿外周血单个核细胞的IFN-γ和IFN-γ/IL-4比值降低,差异有统计学意义(P<0.05);哮喘患儿外周血单个核细胞在体外与头孢呋辛(100 mg/L)孵育48 h后,IL-4水平升高,差异有统计学意义(P<0.05),而IFN-γ的变化无统计学意义(P>0.05);IFN-γ/IL-4比值则降低,差异有统计学意义(P<0.01)。结论哮喘患儿外周血单个核细胞以Th2(IL-4)占优势,Th1/Th2比值平衡失调;而头孢呋辛更加剧这一倾斜,不利于哮喘治疗。     

Cytogenetic damage in blood lymphocytes and exfoliated epithelial cells of children with inflammatory bowel disease

This longitudinal, prospective study sought to establish whether pediatric Crohn's disease (CD) and ulcerative colitis (UC) are associated with increased levels of cytogenetic damage and whether folate supplementation in combination with other treatments mitigates cytogenetic damage in children with inflammatory bowel disease (IBD). After a 1-mo treatment and folate supplementation, all clinical tests in CD (n = 24) and UC (n = 17) patients improved. Patients with CD were comparable in the cytogenetic response with controls (n = 28) assessed by micronucleus (MN) assay, but both groups differed from the UC group. While the MN frequency in epithelial cells slightly decreased from first to second observations in CD patients (p = 0.05) and controls (p = 0.11), an increase was observed in UC patients (p = 0.001). Similar changes were observed in blood lymphocytes resulting in significantly higher levels of the MNs and chromosome bridges in UC patients. These preliminary findings of a difference in chromosome damage between pediatric UC patients compared with CD patients and healthy controls warrant confirmation and expansion to determine (1) the role of cytogenetic damage in the pathogenesis of these diseases, (2) relative contribution of treatment and folate supplementation, and (3) potential links to the eventual development of cancer in some patients.     

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.     

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。     

Expression of very late antigen-4 and lymphocyte function-associated antigen-1 on peripheral blood lymphocytes from patients with graves disease

The aim of the study was to estimate the expression of beta1 integrin CD49d (very late antigen-4), beta2 integrin CD11a (lymphocyte function-associated antigen 1 alpha), L-selectin (CD62L), and intercellular adhesion molecule-1 [ICAM-1 (CD54)] on peripheral blood mononuclear cells in patients with Graves disease (GD) (n = 45, mean age 15.9 +/- 5.9 y), in patients with nontoxic nodular goiter (NTNG) (n = 25, mean age 15.2 +/- 5.7 y), and in sex- and age-matched healthy control subjects (n = 25, mean age 15.9 +/- 2.4 y). The expression of the lymphocyte adhesion molecules was analyzed by the three-color flow cytometry. Decreased percentages of CD49d+ and CD11a+ lymphocytes were observed in untreated Graves' patients in comparison to healthy controls (p < 0.007, p < 0.036) and non-toxic nodular goiter patients (p < 0.006, p < 0.019). The analysis of CD62L+ and CD54+ antigen expression on peripheral blood lymphocytes revealed increased percentages of L-selectin- and ICAM-1-positive cells in patients with GD (p < 0.005 for CD62L, p < 0.008 for ICAM-1) before methimazole therapy, compared with healthy controls. These abnormalities were absent in children with NTNG. In patients with untreated GD, a positive correlation was found between serum levels of free thyroxine and the percentage of CD54+ lymphocytes (r = 0.47, p < 0.036). Statistically significant negative correlations existed between free thyroxine concentration in blood serum and the percentage of CD11a+ lymphocytes (r = -0.39, p < 0.033). No such correlation was detected between the percentage of lymphocyte markers and serum levels of antithyroid antibodies. We conclude that the alterations in the expression of adhesion molecules on peripheral blood mononuclear cells might suggest their role in the development of GD.     

Disruption in cytokine and chemokine production by T-cells in vertically HIV-1-infected children

This study measured cytokine production by mitogen-stimulated peripheral blood mononuclear cells (PBMCs) from 55 human immunodeficiency virus (HIV)-infected children born to HIV-infected mothers, and compared it with vertically exposed but uninfected age-matched children. A significant defect was observed in Th1 cytokine production [interferon-gamma and interleukin-2 (IL-2)] in HIV-infected children compared with controls, but without a concomitant increase in Th2 cytokines. Indeed, IL-5 and IL-10 production was even lower in HIV-infected children than in controls, with the decrease in IL-5 being the best predictive marker of immunodeficiency. In addition, an increased release of tumour necrosis factor-alpha (TNF-alpha) that correlated well with CD4+ levels, and a positive correlation of the TNF-alpha/IL-10 ratio with disease progression was observed. A correlation between AIDS-free status and higher %CD4+ and %CD8+ T-lymphocytes and RANTES (regulated on activation, normal T-cell expressed and secreted) production was also found. Conclusion: A dysfunctional cytokine production of PBMCs was observed in HIV-infected children in both Th1 and Th2 cytokines due to quantitative and qualitative defects induced by HIV-1. An important observation was an increased RANTES production associated with viral isolates of NSI/R5 phenotype and S/L kinetics.     

小儿脓毒症T细胞亚群变化分析及临床意义

目的：通过观察脓毒症患儿T细胞亚群变化及预后,探讨其在脓毒症发生发展中的临床意义。方法：以50 例严重脓毒症患儿和150 例一般脓毒症患儿为研究对象,同时纳入50例非脓毒症儿童为对照组。采用流式细胞仪测定外周血CD3阳性淋巴细胞(CD3+T)、CD4阳性淋巴细胞(CD4+T)和CD8阳性淋巴细胞(CD8+T)百分比及CD4+T／CD8+T比值,并于入院24 h内行小儿危重病例评分（PCIS）。结果：严重脓毒症组患儿CD3+ T、CD4+ T和CD8+ T水平、CD4+ T／CD8+ T比值及PCIS明显低于对照组及一般脓毒症组(P<0.01)。200例脓毒症患儿中,治愈组CD3+ T、CD4+T、CD8+T水平、CD4+T／CD8+T比值及PCIS较死亡组降低(P<0.05)。结论：脓毒症患儿存在着不同程度的细胞免疫功能抑制,其程度与疾病的严重程度有明显的关联。脓毒症患儿外周血T淋巴细胞亚群的检测对评估脓毒症患儿的免疫功能状况和判断病情严重程度具有重要的临床意义。     

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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+...     

Ontogeny of T-helper 1 and T-helper 2 cytokine production in childhood

Compared to adults, infants and young children demonstrate differences in their immune response, indicating that there is maturation or change over time and it is probable that this may be reflected in cytokine production. Cytokine responses have been demonstrated to be different in atopic and non-atopic individuals. In this study, we examined T-helper 1 (Th1) (interferon-γ[IFN-γ]) and T-helper 2 (Th2) (interleukin [IL]-4, IL-5, and IL-13) cytokine release from atopic and non-atopic children in response to the staphylococcal superantigen, staphylococcal enterotoxin B (SEB). In non-atopic and atopic children, IFN-γ, IL-4, and IL-5 release was significantly related to age. Non-atopic children younger than 2 years of age were found to have significantly reduced Th2 (IL-4, IL-5, and IL-13) responses when compared with older, non-atopic children. Atopic children had a reduced IFN-γ response when compared with non-atopics in early childhood; however, the decreased IFN-γ response seen in early childhood did not persist after 10 years. These age-related changes in cytokine production provide further support for the concept that cytokine deviations may determine the natural history of atopic disease during early childhood. In addition, the present study indicates the necessity of age-matched controls when examining children for both Th1 (IFN-γ) and Th2 (IL-4) cytokine release.     

CD4+CD25int/highCD127low 调节性T细胞在再生障碍性贫血患儿中的检测及意义

目的：探讨再生障碍性贫血（AA）患儿外周血 CD4+ CD25int/high CD127low 调节性 T 细胞（Treg）的表达特点及其与 Hb、WBC 和 血小板（Plt）数量的相关性。方法：采用流式细胞术检测 15 例正常儿童（对照组）和 22 例治疗有效的AA患儿治疗前后外周血 CD4+ CD25int/high CD127low Treg百分比的变化,分析 CD4+CD25high-CD127low Treg与 Hb、WBC 和 Plt 数量的相关性。结果：AA 组急性期外周血 CD4+CD25+、CD4+CD25high、CD4+CD25+CD127low、CD4+CD25highCD127low Treg占 CD4+ T 细胞的百分比均较对照组显著降低（P﹤0.05）;恢复期的表达则高于急性期（P﹤0.05）,而与对照组比较差异无统计学意义（P﹥0.05）。AA组急性期、恢复期及对照组的 CD4+CD25highCD127low Treg表达率与外周血中 Hb、WBC 和 Plt 数量均呈正相关（r分别为 0.499、0.526、0.540,P<0.05）。结论：CD4+CD25int/highCD127low Treg 减低可能与 AA 发病有关,为其作为判断 AA 病情变化的指标提供了可能的理论依据,并可以作为免疫治疗的靶点之一进行深入研究。     

炎症性肠病患儿血清Th17相关细胞因子表达及其意义

目的　探讨儿童IBD患者血清中Th17相关细胞因子的表达水平及其临床意义。方法　收集2017年4月至2019年5月在西安交通大学附属儿童医院消化科住院治疗的儿童IBD患者40例,以同期21例非炎症性疾病儿童作为对照组。采集受检者血清标本,通过酶联免疫吸附试验检测白细胞介素-17A（IL-17A）、 IL-17F、 IL-21、 IL-22、 IL-25的表达水平。结果　IL-17A、 IL-17F、 IL-21和IL-22在溃疡性结肠炎（UC）及克罗恩病（CD）患儿血清中表达明显升高,IL-25明显降低（P＜0.05）;IL-17A、 IL-17F及IL-21血清水平与UC患儿疾病活动度呈正相关（P＜0.05）;IL-21在UC患儿血清中的表达与IL-17A和IL-17F的表达呈正相关（P＜0.05）。结论　Th17相关细胞因子在IBD患儿血清中表达失调,为进一步研究Th17细胞在儿童IBD中的作用提供了依据;IL-17A、 IL-17F、 IL-21血清水平与UC患儿疾病活动度呈正相关,表明其可能是Th17细胞触发儿童UC的重要促炎细胞因子。     

Th17/Treg细胞比例失衡在儿童原发性免疫性血小板减少症中的意义

目的 探讨Th17/Treg 细胞比例失衡在儿童原发性免疫性血小板减少症(ITP)发病及治疗中的意义。方法 选取2015 年5 月至2015 年8 月确诊为ITP 的32 例患儿作为ITP 组,同期选取22 例健康儿童作为健康对照组,采用流式细胞术分别检测初诊ITP 患儿、丙种球蛋白治疗后的ITP 患儿和健康对照组儿童外周血Th17、Treg 细胞的比例。结果 ITP 患儿治疗前外周血Th17 占CD4⁺T 细胞的比例、Th17/Treg 细胞比值均显著高于治疗后及健康对照组儿童(P<0.05),治疗前Treg 细胞占CD4⁺T 细胞的比例显著低于治疗后及健康对照组儿童(P<0.05);32 例ITP 患儿经治疗后,20 例完全反应,4 例有效,8 例无效,完全反应患儿外周血Th17细胞占CD4⁺T 细胞比例、Th17/Treg 细胞比值显著低于无效患儿(P<0.05)。结论 儿童ITP 中存在Th17/Treg细胞比例失衡,丙种球蛋白可通过调节Th17/Treg 细胞比例变化进而改变患儿细胞免疫功能,治疗过程中检测该比值变化可能对疾病的疗效有一定的预测作用。     

儿童郎格罕斯细胞组织细胞增生症免疫功能检测意义

目的 了解郎格罕斯细胞组织细胞增生症(LCH)患儿治疗前后细胞和体液免疫功能状态.方法 将30例LCH患儿按Lavin-Osband法分为3组.采用流式细胞仪检测LCH患儿及26例正常对照组儿童外周血CD3+、CD+、CD8+、CD3-CD(16+56)+、CD19+含量,采用特定蛋白分析仪检测患儿及正常对照儿童血清IgA、IgM、IgG含量.结果 三组LCH患儿确诊时,各组间CD3+、CD19+差异均无统计学意义(P均>0.05),Ⅲ组LCH患儿CD4+/CD8+、CD3-CD(16+56)+低于其他组,差异均有统计学意义(P均<0.05);LCH患儿确诊时各组间血清Ig与正常对照组比较差异均无统计学意义(P均>0.05);Ⅱ组和Ⅲ组LCH患儿治疗6个月时CD4+/CD8+分别低于治疗前,差异均有统计学意义(P均<0.05);LCH患儿治疗前后CD3-CD(16+56)+、CD19+比较,差异均无统计学意义(P均>0.05).LCH患儿治疗前后血清Ig比较,差异无统计学意义(P>0.05).结论 LCH患儿发病时存在细胞免疫功能降低,与病情轻重有关;体液免疫功能无明显改变.LCH患儿化疗后可出现细胞免疫功能紊乱加重.检测LCH患儿的免疫功能,可为免疫调节治疗提供客观依据.     

手足口病患儿外周血血管活性肠肽的表达

目的 初步探讨手足口病患儿外周血血管活性肠肽的表达水平和意义。方法 根据病情将86例手足口病患儿分为1期组 （19例）和2期组 （67例）。采用ELISA法检测外周血血浆中血管活性肠肽、γ-干扰素 （IFN-γ）和白细胞介素-4 （IL-4）浓度。流式细胞术检测CD3+T、CD4+T、CD8+T淋巴细胞亚群。RTPCR法定性检测大便肠道病毒71型 （EV71）-RNA。结果 2期组EV71-RNA阳性率明显高于1期组 （P < 0.05）;2期组血清IgG、IgA、IgM、补体C3水平高于1期组 （P < 0.05）;2期组外周血CD3+T、CD4+T、CD8+T淋巴细胞亚群比例低于1期组 （P < 0.05）;2期组外周血B细胞比例高于1期组 （P < 0.05）;2期组CD4+/CD8+比值低于1期组 （P < 0.05）;2期组外周血血管活性肠肽浓度低于1期组 （P < 0.05）。86例手足口病患儿外周血血管活性肠肽浓度与CD4+T淋巴细胞亚群比例以及CD4+/CD8+比值呈正相关 （分别r=0.533、0.532,P < 0.05）。结论 血管活性肠肽可能是反映手足口病严重程度的重要标记物。     

Soluble CD30 and CD23 in cord blood are not related to atopy in early childhood

Atopic disease, including atopic dermatitis (AD), is associated with a T-helper 2 (Th2)-dependent immune response. The cytokine receptor CD30 appears to be preferentially expressed on, and its soluble form (sCD30) released by, Th2 cells. Therefore, sCD30 may be a potential marker for atopic disorders. The aim of this study was to test the hypothesis that the sCD30 level in cord blood could be used to predict the development of atopy or AD in early childhood. In a case-control study, levels of sCD30, as well as soluble low-affinity immunoglobulin E (IgE) receptor (sCD23), interleukin-4 (IL-4) and IgE, were measured in cord blood in 35 children who subsequently developed allergic sensitization and AD before the age of three, and the results were compared to those of 35 matched children without a history of atopy. There was no difference in cord blood levels of sCD30 between controls (32.5 U/ml; 19.7–80.1) and children with subsequent atopy and AD (32.2 U/ml; 22–75.9) (median; quartiles). The concentration of sCD30 showed no relation to the levels of total IgE, sCD23 or IL-4. Levels of sCD23 were similar in children with subsequent atopy (60.2 U/ml; 44.5–76.8) and controls (55.2 U/ml; 38.3–72.5), whereas IL-4 was detectable in 10 of the atopic children and in only two of the controls (p < 0.05). In conclusion, cord blood levels of sCD30 or sCD23 do not seem to be related to the subsequent development of atopy or AD at the age of three.     

Intracellular production of IL-2, IL-4, IFN-γ, and TNF-α by peripheral blood CD3+ and CD4+ T cells in children with atopic dermatitis

The role of the type-2 T helper (Th2) cell-mediated immune response in the immunopathogenesis of atopic dermatitis (AD) is well documented. Whether polarized immunoresponse is confined to antigen-specific T cells or is distributed among all T cell subsets is still controversial. We investigated frequencies of interleukin-2 (IL-2), IL-4, interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) producing CD3⁺ and CD4⁺ T cells in peripheral blood from children with atopic dermatitis and healthy subjects with and without in vitro stimulation. Children with severe AD had a significantly lower percentage of CD4⁺ T cells spontaneously expressing IL-4 compared with healthy controls (p <0.01). Polyclonal stimulation significantly increased cytokine production in both AD patients and healthy individuals. Frequencies of CD3⁺ and CD4⁺ producing IL-2, IL-4, IFN-γ, and TNF-α after in vitro stimulation with phorbol-12-myristate 13-acetate (PMA) + ionomycin were comparable in the AD and control groups. In response to PMA/ionomycin, children with AD and asthma symptoms had a significantly lower percentage of CD3⁺ T cells producing TNF-α. We failed to demonstrate evidence of an imbalance with respect to type-2 cytokine productions in children with AD. Comparable induction of Th1 and Th2 cytokines in polyclonally stimulated peripheral CD3⁺ and CD4⁺T cells from AD patients and controls puts into question the polarized Th2 immune response as a general characteristic of T cells in children with atopic dermatitis.