CXCR4在儿童急性白血病的表达及临床意义

目的探讨儿童急性白血病(AL)骨髓细胞表面CXCR4的表达及其临床意义。方法采用流式细胞术分别检测43例初诊未治急性白血病患儿(实验组)和10例非恶性血液病患儿(对照组)骨髓细胞表面CXCR4的表达情况。结果1.急性白血病患儿骨髓细胞表面CXCR4的相对荧光强度明显高于对照组(P<0.05)。2.ALL组CXCR4的相对荧光强度明显高于AML组(P<0.05)。3.髓外浸润组患儿初诊时CXCR4的相对荧光强度明显高于非髓外浸润组。(P<0.05)。4.实验组患儿CXCR4的相对荧光强度与外周血白细胞计数(WBC)呈正相关关系,相关系数为0.58(P<0.05)。结论1.CXCR4在儿童急性白血病初诊时为高水平表达,提示它可以作为急性白血病的一种检测指标。2.CXCR4的表达在ALL明显高于AML,说明它与急性白血病的类型具有一定相关性。3.急性白血病患儿骨髓细胞表面CXCR4的高表达与髓外浸润密切相关,并且和初诊时高白细胞计数有关。     

Serum concentrations of CCR4 ligands in relation to clinical severity of atopic dermatitis in Egyptian children

T helper 2 (Th2) lymphocytes, the key effector cells in pathogenesis of atopic dermatitis (AD), express CCR4 receptors. CCR4 ligands (macrophage-derived chemokine ‘MDC’ and thymus and activation-regulated chemokine ‘TARC’) direct trafficking and recruitment of Th2 cells into lesional skin in AD. These chemokines appear to be useful inflammatory markers for assessing severity of AD in adults. However, the same results have not been replicated in children. Therefore, we were stimulated to elucidate the expression of CCR4 ligands in children with AD and their relation to clinical disease severity. To investigate this, serum concentrations of CCR4 ligands were determined in 60 children, of whom 30 had AD and 30 were healthy matched subjects. Patients were classified into mild (n = 8), moderate (n = 12) and severe (n = 10) according to the objective scoring AD (obj-SCORAD) index. Serum concentrations of MDC and TARC were significantly increased in children with AD (2697 ± 982.6 pg/ml and 945.5 ± 494.7 pg/ml, respectively) compared with controls (357.2 ± 233.2 pg/ml and 214.2 ± 116.6 pg/ml, respectively, p < 0.0001). Serum levels of both chemokines went hand in hand with disease severity as they were significantly higher in severe than moderate and in moderate than mild AD. In addition, they correlated positively with obj-SCORAD (r = 0.99 for both, p < 0.0001). Furthermore, both chemokines had significant positive correlations to blood eosinophil counts and serum immunoglobulin E. In conclusion, serum CCR4 ligands may be useful inflammatory markers for assessing AD severity in children. Further studies may pave way for CCR4 ligands antagonism among the adjuvant therapeutic strategies of AD.     

小鼠骨髓间充质干细胞对植物血凝素诱导小鼠脾细胞增殖后趋化因子受体表达的影响

目的：观察小鼠骨髓间充质干细胞（mesenchymal stem cells,MSCs）在体外对小鼠脾细胞表达趋化因子受体的影响。方法：用密度梯度离心法从小鼠骨髓中分离出小鼠骨髓间充质干细胞,经低糖DMEM培养基培养扩增。C57BL/6小鼠脾细胞以1×106/孔的密度接种于24孔板,加入植物血凝素(PHA),培养72 h。实验分3组：A组按10%比例加入MSC;B组按1%比例加入MSC;对照组不加MSC。3 d后收集悬浮的脾细胞进行流式细胞术检测小鼠脾细胞趋化因子受体CXCR3,CCR5,CCR7表达的变化。结果：CD3+CCR5+、CD3+CCR7+在A,B组及对照组中表达的差异均有统计学意义（均P<0.01）;以A组表达率最高,B组次之,对照组最低;CD3+CXCR3+细胞在A组中表达较B组、对照组高（P<0.05）,B组和对照组之间的表达差异无统计学意义。结论：骨髓间充质干细胞在一定浓度下对小鼠脾细胞增殖后趋化因子受体CXCR3,CCR5,CCR7表达有上调作用。［中国当代儿科杂志,2007,9（6）：571-573］     

毛细支气管炎患儿外周血淋巴细胞CXCR3及IP-10的表达及意义

目的 探讨趋化因子受体3(CXCR3)及γ干扰素诱导蛋白-10(IP-10)在毛细支气管炎(简称毛支)患儿外周血中的表达及临床意义.方法 随机选取毛支住院患儿55例,按有无过敏因素分为毛支Ⅰ组(有过敏因素)和毛支Ⅱ组(无过敏因素);同期住院的外科非感染患儿28例作为对照组.采用流式细胞术检测3组患儿外周血CD4+、CD8+淋巴细胞表面CXCR3(表面分子标记为CD183)的表达,ELISA 法测定血清中其配体IP-10的水平.结果 毛支Ⅰ组和毛支Ⅱ组外周血CD4+ T细胞表面CD183+细胞的表达及CD8+ T细胞表面CD183+细胞的表达均高于对照组(PPP结论 CXCR3及IP-10参与了毛支的发病过程,且CXCR3与过敏因素有关.     

T lymphocytes in food allergy: Overview of an intricate network of circulating and organ‐resident cells

Although food hypersensitivity might be divided in IgE‐ and non‐IgE mediated food allergy, there is a large body of evidence implicating T lymphocytes overall in the pathogenesis of food allergy. Priming of naive T cells will occur mainly in Peyer's patches (PP), where surface receptors (l ‐selectin, CCR7 and CXCR4) will help to initiate diapedesis of the cells to the submucosa. Various antigen‐presenting cells (e.g. dendritic cells, M cells) will present food antigen‐derived epitopes and initiate either non‐responsiveness, or a food‐mediated immune response. Food‐specific memory T cells express various surface receptors such as the α4β7‐integrin, or the cutaneous lymphocyte antigen. It is speculated, that they might also express specific chemokine receptors (CCR4, CCR7 or CCR9). Organ‐specific homing will be facilitated through the corresponding receptors (i.e. MAdCAM‐1 in the gut, VCAM‐1 or fibronectin in other mucosal organs, or E‐selectin in the skin). Locally secreted chemokines might help to attract T cells through their corresponding chemokine receptors. Finally, potential T‐cell directed therapeutic interventions (peptide‐derived immunotherapy, DNA vaccination, or strategies preventing T cells from trafficking to target organs) are discussed.     

Chemokine IL-8 and chemokine receptor CXCR3 and CXCR4 gene expression in childhood acute lymphoblastic leukemia at first relapse

In this study, we examined the gene expression of interleukin (IL)-8, CXCR3, and CXCR4 in leukemic cells from 100 children with relapsed B-cell progenitors (BCP) acute lymphoblastic leukemia (ALL), using quantitative real-time polymerase chain reaction (RT-PCR). IL-8, CXCR3, and CXCR4 were expressed in almost all bone marrow (BM) samples. The CXCR4 expression significantly correlated with known prognostic factors at relapse: time point and site of relapse. Patients who had a combined BM relapse (n=21) had lower IL-8 and CXCR4 expression than those who had an isolated BM relapse (n=79). The CXCR3 expression was higher in female patients (n=39) than in male patients (n=61). However, this did not reach prognostic relevance in relapsed ALL.     

环磷酰胺和吡柔比星对横纹肌肉瘤RD细胞株CXCR4表达的影响

目的观察敏感剂量下环磷酰胺（CTX）和吡柔比星（THP）对体外培养人横纹肌肉瘤RD细胞株CXCR4基因表达的影响。方法体外培养RD细胞株;MTT试验明确CTX和THP对RD细胞的效应剂量;细胞划痕实验检测RD细胞的迁移能力;RT—PCR及Westernblot法检测RD细胞CXCR4表达。结果（1）敏感剂量下CTX（30mmol／L）和THP（1000ng／mL）均可抑制RD细胞的迁移（P〈0．05）,两药联合应用对细胞迁移抑制作用增强,较对照组差异有非常显著性（P〈0．01）;（2）各药物处理组（CTX,THP,CTX＋THP）药物作用于RD细胞24h后CXCR4蛋白的表达较对照组减少,其差异均有显著性（P〈0．05）;其中CTX＋THP组较CTX组CXCR4蛋白表达减少更明显（P〈0．05）,但与THP组比较,差异无显著性（P〉0．05）;（3）同样条件下,各药物处理组（CTX．THP,CTX＋THP）与对照组比较,RD细胞CXCR4mRNA的表达均减少,差异均有显著性（P〈0．05）;CTX＋THP组较CTX组CXCR4mRNA表达减少明显（P〈0．05）,但较THP组差异无显著性（P〉0．05）。结论化疗药物CTX和THP均能抑制人横纹肌肉瘤RD细胞的增殖;敏感剂量下的两种药物均能抑制RD细胞的迁移及下调RD细胞CXCR4的表达,提示两种化疗药物发挥作用的机制可能涉及到CXCR4基因。     

Chemokine receptor CCR2 and CCR5 polymorphisms in children with insulin-dependent diabetes mellitus.

Studies have shown the important roles of several regulatory and proinflammatory cytokines in insulin-dependent diabetes mellitus (IDDM). CC-chemokine receptors CCR2 and CCR5 bind chemokines that are involved in the trafficking of leukocytes in both basal and inflammatory states. A common 32-bp deletion mutation in the CCR5 gene (CCR5delta32) and a G-to-A nucleotide substitution in the CCR2 at position 190 (CCR2-64I) have recently been described. In the present study, we have determined the frequency of the CCR5delta32 and CCR2-64I alleles in children with IDDM [n = 115; age 1-14 (9.3+/-4.3) y] and in nondiabetic subjects [n = 280; age 1-14 (8.5+/-4.5) y]. The CCR5delta32 allele frequencies were 0.117 in children with IDDM and 0.111 in nondiabetic subjects, indicating that the deletion allele has no association with IDDM. The CCR2-64I allele frequency in children with IDDM was 0.226, which differed significantly from the allele frequency in controls (0.114, p = 0.001). The role of this mutation in IDDM cannot be explained yet, but, because CCR2 mediates the chemotaxis of CD4+ and CD8+ T cells to areas of inflammation and because these cells play important roles in insulitis, a mutation in the CCR2 gene may contribute to the susceptibility to the disease. Alternatively, the 64I allele could be a marker of a linked mutation through linkage disequilibrium. According to these results, the CCR2 gene may be a new candidate for the susceptibility locus of IDDM. However, because no IDDM locus has been identified near 3p21 until now, further investigations are needed to confirm this statement.     

不同转移潜能人神经母细胞瘤细胞系趋化因子受体CXCR4的表达及对瘤细胞趋化作用的影响

目的 以自建的人神经母细胞瘤转移瘤(QDDQ-NM1)及原位瘤(QDDQ-NM2)细胞系的细胞为研究对象,探讨CXCR4在不同转移潜能人神经母细胞瘤细胞系细胞中的表达情况及其对瘤细胞趋化作用的影响.方法 采用RT-PCR检测两神经母细胞瘤细胞系细胞CXCR4 mRNA的表达;流式细胞仪直接免疫荧光法检测两细胞系细胞膜表面CXCR4蛋白的表达;通过趋化和趋化抑制实验观察CXCR4特异性配体CXCL12对QDDQ-NM1和QDDQ-NM2细胞系瘤细胞的趋化作用.结果 RT-PCR显示QDDQ-NM1细胞系CXCR4mRNA的相对含量为0.57±0.08,QDDQ-NM2细胞系为0.29±0.05,两者比较差异有统计学意义(P=0.005＜0.05),流式细胞仪直接免疫荧光法检测两细胞系细胞膜表面CXCR4蛋白表达的阳性率分别为(64.59±1.57)%和(36.72±0.57)%,两者的差异有统计学意义(P=0.00＜0.05),CXCR4特异性配体CXCL12可在一定范围内呈浓度依赖性的趋化QDDQ-NM1和QDDQ-NM2细胞系细胞的迁移,以100 ng/ml的效果较为明显,两细胞系迁移到聚碳酸酯膜下的细胞数差异有统计学意义(P＜0.05).CXCR4特异性拮抗剂AMD3100能有效抑制这种趋化作用(P＜0.05).结论 QDDQ-NM1细胞系的细胞功能性高表达趋化因子受体CXCR4,可能与人神经母细胞瘤QDDQ-NM1细胞系细胞的体外高转移潜能有关.

Abstract：

Objective To study the expression of functional chemokine receptor CXCR4 and its effects on the metastatic potential of human neuroblastoma.Methods Two human neuroblastoma cell lines were used in this study,including QDDQ-NM1 with high metastatic potential and QDDQ-NM2 with low metastatic potential.The expression of CXCR4 was explored at mRNA level using RT-PCR,and the protein level by flow cytometry.Chemotaxis assay was also performed to study the migratory response of QDDQ-NM1 and QDDQ-NM2 to CXCR4 ligand CXCL12.Results The mRNA of CXCR4 was higher in QDDQ-NM1 group than that in QDDQ-NM2 group (0.57±0.08 vs 0.29±0.05,P =0.005).The expression of CXCR4 on QDDQ-NM1 group was also higher than that in QDDQ-NM2 group [(64.59 ±1.57) % vs (36.72 ± 0.57) %,P＜0.05] The QDDQ-NM1 cells exhibited stronger migratory response to CXCL12 in a concentration dependent manner (P＜0.05),and the response peaked to CXCL12 at 100 ng/ml.AMD3100,a specific CXCR4 antagonist,could reverse the tumor's migratory response to CXCL12.Conclusions The expression of CXCR4 is associated with the metastatic potential of human neuroblastoma.     

Primary isolated myeolosarcoma in childhood]

Isolated myelosarcomas are rare first manifestations of acute myeloid leukemia (AML), preceding bone marrow involvement by weeks to months. Seventeen of 654 children observed during the studies AML-BFM 87 and 93 were diagnosed as extramedullar myelosarcomas (2.6%). The predominantly myelomonocytic or monoblastic tumor cells (M4 or M5 according to FAB classification) mainly infiltrated skin (n = 8). Additional tumors were located in mucosa (n = 2), central nervous system (n = 2), orbita (n = 2), bone (n = 1), glandulae parotis (n = 1) and lymph nodes. Due to the initial mild and variable symptoms in some children the diagnostic measurements were delayed and treatment was inadequate. This might be responsible for the high rate of relapse (79%) and the poor outcome. Ten of 17 patients died from disease (estimated survival 0.27 +/- 0.13 compared to AML-BFM 87/93 0.51 +/- 0.03). Suspect skin lesions or tumors should be considered as isolated myelosarcoma of a primary manifestation of AML. An intensive AML-specific chemotherapy is recommended.     

CC chemokine receptor expression in childhood asthma is influenced by natural allergen exposure

Chemokines and their receptors may play an important role for leukocyte trafficking in allergic inflammation. Aim was to evaluate whether expression of chemokine receptors CCR4 and CCR8 on cells obtained by sputum induction from asthmatic allergic children may be influenced by house dust mite (HDM) allergen natural exposure. Twenty-one children (7-13 yr) with moderate asthma and sensitized to HDM were evaluated during a prolonged period of allergen avoidance (T0) and after a period of natural allergen exposure (T1). At each time point of sputum induction, lung function evaluation, exhaled nitric oxide (eNO) measurements were performed. At T1, CCR4 and CCR8 expression on sputum-induced cells increased from 28.4% +/- 2.9% and 25.8% +/- 1.9%, to 41.1% +/- 4.2% and 37.5% +/- 2.0%, respectively (p < 0.05 and p = 0.01). After allergen exposure, both sputum eosinophils (from 5.2% +/- 2.0% to 12.1% +/- 4.1%, p < 0.01) and eNO (from 15.1 +/- 2.2 ppb to 24.2 +/- 5.8 ppb, p < 0.05) showed significant increase. Lung function tests presented significant deterioration of Forced Expiratory Flow at 25-75% of Vital Capacity (FEF(25--75)) (p < 0.05) and increase of residual volume (p = 0.002). Significant changes in CC chemokine receptor expression in sputum-induced cells in asthmatic children in response to HDM exposure have been observed leading to consider the relevance of CCR4 and CCR8 in allergic asthmatic inflammation.     

CCR2基因rs1799864多态性与儿童噬血性淋巴组织细胞增生症的关联研究

目的 探讨CCR2基因单核苷酸多态性(SNP)位点V64I(rs1799864)与儿童噬血性淋巴组织细胞增生症(HLH)发病的关联性.方法 收集2007年1月至2013年12月确诊为HLH的86例患儿的临床资料,采用SNaPshot基因分型技术对HLH患儿和128例健康对照进行CCR2基因rs1799864位点分型,比较两组该SNP位点基因型和等位基因频率的差异;以及HLH患儿不同临床特征与rs1799864位点基因型分布的关系.结果 与对照组相比,HLH组rs1799864位点的基因型和等位基因频率差异均无统计学意义(均P>0.05);就发病年龄是否<1岁、治疗后1 d体温是否恢复正常及治疗后2~3周血小板是否恢复正常等不同临床特征的HLH患儿基因型分布进行比较后发现差异均具有统计学意义(均P<0.05).结论 CCR2基因 rs1799864位点多态性与儿童HLH的发病无关,但其基因型不同可能与HLH患儿的临床表现及预后有关.     

RNA干扰趋化因子受体4基因表达对神经母细胞瘤体外侵袭能力的影响

目的 构建趋化因子受体4(CXCR4)小分子干扰RNA(siRNA)表达载体,研究其对体外神经母细胞瘤侵袭能力的影响.方法 选择CXCR4高表达的神经母细胞瘤SH-SY5Y细胞系,设计合成人CXCR4基因不同靶点的能编码siRNA的3条双链DNA序列,克隆到真核表达载体pSilencerTMneo中构建siRNA表达载体,体外脂质体介导转染SH-SY5Y细胞,用半定量RT-PCR分析CXCR4基因mRNA的变化,用免疫组织化学和Western blot分析CXCR4蛋白表达,Transwell小室检测细胞的侵袭能力.结果 成功构建了CXCR4-siRNA表达载体,转染后半定量RT-PCR检测神经母细胞瘤细胞CXCR4 mRNA丰度分别为siR1转染组0.32±0.09、siR2转染组0.35±0.13和siR3转染组0.33±0.11,相对于对照组0.58±0.13表达下降(P＜0.05);转染后免疫组化检测神经母细胞瘤细胞CXCR4的蛋白表达分别为siR1转染组75.98±4.81、siR2转染组75.52±3.95和siR3转染组76.35±6.51,相对于对照组92.196±3.89表达下降(P＜0.01);转染后Western b1ot检测神经母细胞瘤细胞CXCR4的蛋白表达分别为siR1转染组0.1103±0.0023、siR2转染组0.1203±0.015和siR3转染组0.1308±0.0018,相对于对照组0.4832±0.0012表达下降(P＜0.01);且转染后神经母细胞瘤细胞侵袭能力较对照组53.11±3.72降低(P＜0.05),siR1转染组为25.48±2.81、siR2转染组为30.89±2.77、siR3转染组为18.83±1.79.结论 CXCR4-siRNA表达载体通过降低CXCR4基因的蛋白表达能显著抑制神经母细胞瘤细胞的体外侵袭能力,有望为神经母细胞瘤的基因治疗开辟新途径.

Abstract：

Objective To explore the effect of silencing chemokine receptor type 4 (CXCR4) by siRNA on the invasion capability of neuroblastoma cell line SH-SY5Y in vitro. Methods Three siRNAs targeting CXCR4 were chemically synthesized and transfected into SH-SY5Y cells. The transfection efficiency was observed under fluorescence microscope. CXCR4 expression at mRNA and protein levels were detected by semi-quantitative RT-PCR and Western blotting. The invasion capability of the cells was evaluated by Boyden Chamber in vitro. Results Compared with control groups, after the SH-SY5Y cells being transfeeted with the three CXCR4 targeting siRNAs, CXCR4 mRNA in transfected cells significantly decreased (0. 32 ± 0. 09, 0. 35 ± 0. 13 and 0. 33 ± 0. 11 vs 0. 58 ± 0. 13, P＜0. 05 ), CXCR4 protein detected by immunohistochemistry was decreased (75. 98 ± 4. 81, 75. 52 ± 3. 95and 76. 35 ± 6. 51 vs 92. 196 ± 3. 89, P＜0. 01 ), CXCR4 protein detected by Western blotting was also decreased (0. 1103 ± 0. 0023, 0. 1203 ± 0. 0015 and 0. 1308 ± 0. 0018 vs 0. 4832 ± 0. 0012, P＜0. 01 ).The invasion capability of the SH-SY5Y cells was decreased 48 hours after the cells were transfected (25.48±2.81, 30.89±2.77 and 18.83± 1.79 vs 53. 11 ±3.72, P＜0.05). Conclusions Silencing CXCR4 by siRNA decreases the invasion capability of SH-SY5Y cells.     

Induction of MCP1, CCR2, and iNOS expression in THP-1 macrophages by serum of children late after Kawasaki disease

Evidence of premature atherosclerosis late after Kawasaki disease (KD) is accumulating. Given the potential roles of monocyte chemoattractant protein-1 (MCP-1), chemokine receptor CCR-2, and inducible nitric oxide synthase (iNOS) in atherogenesis, we sought to determine whether serum obtained from children late after KD would induce expression of these genes in macrophages in vitro. A total of 79 subjects were studied, which comprised 57 KD patients, 33 of whom had coronary aneurysms, and 22 age-matched controls. Expression of MCP-1, CCR2, and iNOS mRNA in THP-1 macrophages in the presence of patient and control serum was quantified as a ratio to beta-actin mRNA and expressed as a percentage of control. MCP-1 expression was significantly increased in the presence of serum from patients with coronary aneurysms. Expression of CCR2 and iNOS was significantly increased when THP-1 macrophages were incubated with serum from patients with and without coronary aneurysms. The magnitude of induction of MCP-1, CCR2, and iNOS or in combinations correlated positively with serum high-sensitivity C-reactive protein (hs-CRP), and low-density lipoprotein (LDL) cholesterol levels and negatively with high-density lipoprotein (HDL) cholesterol level. In conclusion, the serum of patients with a history of KD induces expression of MCP-1, CCR2, and iNOS in THP-1 macrophages in vitro. Induction of these genes in vivo may be related to chronic inflammation and may have important implications for premature atherosclerosis.     

Comparison of childhood myelodysplastic syndrome, AML FAB M6 or M7, CCG 2891: report from the Children's Oncology Group

BACKGROUND: Myelodysplastic syndromes (MDS), acute erythroleukemia (FAB M6), and acute megakaryocytic leukemia (FAB M7) have overlapping features. PROCEDURE: Children without Down syndrome or acute promyelocytic leukemia who were newly diagnosed with primary myelodysplastic syndrome or acute myeloid leukemia (AML) M6 or M7 were compared to children with de novo AML M0-M5. All children were entered on the Children's Cancer Group therapeutic research study CCG 2891. RESULTS: The presentation and outcomes of the 132 children diagnosed with MDS (60 children), AML FAB M6 (19 children), or AML FAB M7 (53 children) were similar. Children with AML FAB M7 were diagnosed at a significantly younger age (P = 0.001). Children with MDS, M6, or M7 had significantly lower white blood cell (WBC) counts (P = 0.001), lower peripheral blast counts (P < 0.001), and an increased frequency of -7/7q- (P = 0.003) at presentation. All three groups had significantly inferior overall survival (OS) (P < 0.001) and event free survival (P < 0.001) compared with the 748 children diagnosed with AML FAB M0-M5 when assessed from entry on study. This poor survival was largely attributable to induction death and failure. However, when assessed from successful completion of induction therapy, the 5-year OS (P = 0.090)(49.1 vs. 56.9%) and disease-free survival (DFS) (P = 0.113)(38.0 vs. 46.3%) therapy were not significantly different from other children with AML. CONCLUSIONS: Childhood AML FAB M6 and AML M7 resemble MDS in presentation, poor induction success rates, and outcomes.     

Understanding the bone marrow microenvironment in hematologic malignancies: A focus on chemokine,integrin, and extracellular vesicle signaling

Signaling between leukemia cells and nonhematopoietic cells in the bone marrow microenvironment contributes to leukemia cell growth and survival. This complicated extrinsic mechanism of chemotherapy resistance relies on a number of pathways and factors, some of which have yet to be determined. Research on cell–cell crosstalk the bone marrow microenvironment in acute leukemia was presented at the 2016 annual Therapeutic Advances in Childhood Leukemia (TACL) investigator meeting. This review summarizes the mini-symposium proceedings and focuses on chemokine signaling via the cell surface receptor CXCR4, adhesion molecule signaling via integrin α4, and crosstalk between leukemia cells and the bone marrow microenvironment that is mediated through extracellular vesicles.     

Analysis of costimulatory molecules OX40/4-1BB (CD134/CD137) detection on chosen mononuclear cells in children and adolescents with Graves' disease during methimazole therapy

Antibody synthesis follows interactions between the T cell receptor (TCR) on activated T lymphocytes and the main histocompatibility complex (MHC) present on APC cells, resulting in lymphocyte proliferation, as well as cytokine synthesis and release. The involvement of costimulatory markers OX40/4-1BB/4-1BBL leads to the enhancement of signals which are necessary for lymphocyte activation in addition to the antigen-specific signal and may prevent anergy. The aim of this study was to estimate the expression of OX40 and 4-1BB molecules on peripheral blood cells in patients with Graves' disease (GD) (n = 35, mean age 16.5 +/- 6.1 years) and non-toxic nodular goiter (NTNG) (n = 35, mean age 16.2 +/- 4.7 years), in comparison with sex- and age-matched healthy controls (n = 35, mean age 16.2 +/- 2.1 years). Expression of the costimulatory molecules on mononuclear cells was analyzed by three-color flow cytometry using a Coulter EPICS XL cytometer. Stimulating and blocking antibodies to the TSH-receptor using JPO9 CHO cells in unfractionated serum were measured by a highly sensitive commercial radioimmunoassay. The analysis of OX40/4-1BB expression in patients with newly recognized Graves' disease revealed a statistically significant increase in the percentage of CD134+ T cells (7% vs 1.4%, p <0.001) and CD137+ T cells (3.2% vs 0.8%, p <0.04) compared to the control group. After 2-6 months of methimazole therapy, the percentage of these cells in the peripheral blood of hyperthyroid patients returned to normal values. In addition, the expression of 4-1BBL (CD137L) was detected only on the surface of active monocytes in patients with untreated GD (3.8%), while in the group with nodular goiter and controls the values were trace (0.6% and 0.2%, respectively). We conclude that the changes of expression of costimulatory molecules on the surface of peripheral blood T cells and their significant relationship with the level of antithyroid antibodies indicate an involvement of these molecules in the pathogenesis of Graves' disease. A marked increase in the percentage of CD134/ CD137+ T cells at disease onset may indicate the need for more aggressive therapy in Graves' disease and for a greater duration than the standard 3-year period.     

川崎病患儿外周血单个核细胞基质细胞衍生因子-1和其受体的变化及意义

目的：观察川崎病(KD)患者血浆基质细胞衍生因子1(SDF1)水平及外周血单个核细胞(PBMC)中SDF1的受体CXCR4mRNA表达变化,并分析其与KD及冠状动脉损害的相关关系。方法：采用ELISA方法及荧光定量PCR技术分别测定12例合并冠脉损害和44例无冠脉损害KD患者的急性期与缓解期血浆SDF1浓度及PBMC中CXCR4mRNA表达变化,并与60例正常人比较。结果：KD患者急性期血浆SDF1蛋白水平为1833±395ng/L;PBMC中CXCR4mRNA表达水平为6.57±2.81较对照组升高(P<0.05,P<0.01);缓解期血浆SDF1蛋白和CXCR4mRNA表达水平较急性期下降(P<0.01),但仍显著高于对照组。KD患者合并冠脉损害者PBMC中CXCR4mRNA表达为8.19±2.39,显著高于无冠脉损害者的6.13±2.77(P<0.01)。结论：KD患者血浆SDF1水平及PBMC中CXCR4mRNA表达是增高的,有可能作为判断KD病情活动状态及冠脉损害的免疫学指标。     

Allogeneic versus autologous versus peripheral stem cell transplantation in CR1 pediatric AML patients: a single center experience

BACKGROUND: Treatment of childhood acute myelocytic leukemia (AML) in first remission, is still evolving. Allogeneic bone marrow transplantation (BMT) in patients with a donor has been well established, but the role of autologous transplantation remains of interest, particularly in the light of some encouraging results in adults. PROCEDURE: Out of 81 pediatric patients with AML in first CR, 67 were biologically randomized for allogeneic (n = 31), autologous (n = 20), or peripheral stem cell transplant (n = 16) after completing consolidation treatment, with the remaining (n = 11) dropping out or receiving chemotherapy. Disease free survival (DFS) of these different groups were analyzed. RESULTS: Allogeneic transplantation is not superior to autologous and autologous peripheral blood stem cell transplantation (PBSCT) (DFS in 5 years is 61%, 50%, and 75%). The 5 years DFS in the autologous PBSCT group is significantly better than in the autologous BMT group (75% vs. 50%, P < 0.05). CONCLUSION: In pediatric AML patients without a donor, autologous BMT or autologous PBSCT appears to be an effective treatment option with low transplant related mortality especially in less privileged countries where the chemotherapy only results are still low.