粒-巨噬细胞集落刺激因子联合白细胞介素-2活化的脐血造血细胞抗肿瘤作用的研究

为探讨粒 巨噬细胞集落刺激因子 (GM CSF)联合白细胞介素 2 (IL 2 )活化的脐血造血细胞体外杀伤白血病细胞株HL 6 0细胞的作用 ,以及对脐血粒 -巨噬造血祖细胞 (CFU GM)生成活性的影响 ,采用MTT比色法测定经GM CSF和 (或 )IL 2激活的脐血造血细胞的抗肿瘤活性及半固体培养法检测活化脐血CFU GM生成活性。结果显示 ,IL 2或GM CSF +IL 2组均可活化脐血产生对体外培养的白血病细胞株HL 6 0细胞的杀伤作用 ,杀伤率分别为 ( 6 7 2± 1 5 1 % )和 ( 6 3 4± 2 3 9% ) ;GM CSF单独应用不能活化脐血产生对白血病细胞株HL 6 0细胞的杀伤作用 :GM CSF或GM CSF +IL 2均可扩增脐血CFU GM ,其产率分别为 ( 1 36 1± 4 2 9) / 1 0 5MNC和 ( 90 1±30 3) / 1 0 5MNC ;IL 2单独应用对CFU GM产率有减少的趋势。结果表明 ,IL 2或GM CSF +IL 2均能活化脐血产生明显的抗肿瘤活性 ;GM CSF或GM CSF +IL 2组使脐血CFU GM得到了明显的扩增 :脐血造血祖细胞治疗和免疫治疗相结合将是肿瘤高剂量化疗中一种有希望的治疗方法。     

粒细胞-巨噬细胞集落刺激因子的研究进展

粒细胞一巨噬细胞集落刺激因子（GM—CSF）主要由体内激活的T细胞、单核巨噬细胞等产生。近年研究发现它是一种有广泛免疫活性的效应因子,在激活免疫反应中起重要作用,参与特异性抗体的产生。GM—CSF主要促进粒、巨噬细胞增殖、分化及功能成熟,提高宿主的防御能力,而且可以活化巨噬细胞产生依赖性细胞介导的细胞毒效应（ADCC）和多形性中性粒细胞（PMN）产生补体介导的吞噬作用,以及增强PMN的各种功能。GM—CSF还能提高C3bi及Fcr受体的表达,现就其近年来的研究进展作一综述。     

粒细胞、粒巨噬细胞集落刺激因子在新生儿期的应用

新生儿感染的发病率及病死率较高，吞噬细胞系统不成熟是主要原因这一，粒细胞，粒巨噬细胞集落刺激因子可刺激粒子巨噬细胞增殖，分化、增强中性粒细胞和单核巨噬细胞功能，临床上应用重组人类集落刺激因子治疗和预防新生儿感染取得了令人鼓舞的效果。     

粒细胞、粒巨噬细胞集落刺激因子在新生儿期的应用

新生儿感染的发病率及病死率较高，吞噬细胞系统不成熟是主要原因之一。粒细胞、粒巨噬细胞集落刺激因子可刺激粒巨噬细胞增殖、分化，增强中性粒细胞和单核巨噬细胞功能。临床上应用重组人类集落刺激因子治疗和预防新生儿感染取得了令人鼓舞的效果。     

新生儿感染性疾病血清白细胞介素-8和粒细胞集落刺激因子水平研究

本文测定96例新生儿感染患儿血清白细胞介素-8(IL-8)和粒细胞集落刺激因子(G-CSF),并与正常新生儿对照组比较。结果表明:感染急性期IL-8与G-CSF均高于恢复期及对照组,对照组与恢复期间无显著差异。此外,中性粒细胞计数与G-CSF存在正相关,而与IL-8无相关性。本文结果提示:IL-8和G-CSF作为炎症介质参与了新生儿感染疾病过程,是反映新生儿感染的重要指标。     

正常足月儿脐血粒细胞集落刺激因子水平与白细胞的相关性

目的 研究正常足月儿脐血粒细胞集落刺激因子（G-CSF）水平与白细胞数、中性粒细胞数及分化抗原CD13、CD33表达的相关性。方法 采用ELISA法测定了33例正常足月儿脐血G-CSF水平，并用Counter Electronics仪计数了同一标本脐血白细胞和中性粒细胞数，用免疫荧光法计数了同一标本脐血CD13、CD33阳性细胞数。结果 21例脐血G-CSF检测阳性标本，经统计学处理后发现G-CSF水平与其他4项均无明显相关性。结论 G-CSF与白细胞增殖分化之间并不是简单而唯一的调节关系，可能有其他细胞因子参与调节。     

白细胞介素15和粒-单核集落刺激因子激活的自然杀伤T细胞对神经母细胞瘤细胞的杀伤作用

目的：研究白细胞介素15(IL-15)和粒-单核集落刺激因子(GM-CSF)对健康儿童自然杀伤T细胞(NKT)增殖和对NKT细胞介导的神经母细胞瘤(NB)细胞的细胞毒作用的影响。方法：采集分离健康儿童外周血T细胞,分别加入IL-15,GM-CSF培养,MTT法检测T细胞增殖程度的变化,流式细胞仪检测NKT/T比例并分选出NKT细胞,MTT法检测NKT细胞对NB细胞毒作用。结果：IL-15刺激组、GM-CSF刺激组和IL-15+GM-CSF刺激组T细胞增殖程度分别是对照组的 1.25 倍、1.2 倍和 1.4 倍(均P<0.01);NKT/T比例分别是对照组的 2.56 倍、3.27 倍和 4.39 倍(P<0.01或0.05);GM-CSF 组和IL-15+GM-CSF组NKT细胞毒作用高于对照组(均P<0.01),IL-15组NKT细胞毒作用与对照组比较差异无显著性(P>0.05)。与GM-CSF组比较,IL-15+GM-CSF组T细胞增殖程度增高但NKT/T比例和NKT细胞毒作用差异无显著性。与IL-15组比较,IL-15+GM-CSF组NKT/T比例和NKT细胞毒作用增高但T细胞增殖程度差异无显著性。IL-15组与GM-CSF组T细胞增殖程度、NKT/T比例和细胞毒作用差异无显著性。结论：NKT细胞对NB具有细胞毒作用,IL-15和GM-CSF联合应用可以促进健康儿童外周血NKT细胞增殖,增强NKT细胞对NB细胞毒作用。     

重组人粒-巨噬细胞集落刺激因子治疗儿童药物性粒细胞缺乏症18例

重组人粒 巨噬细胞集落刺激因子 (ｒＨｕＧＭ ＧＳＦ)能特异性刺激粒细胞和巨噬细胞的分化、增殖和成熟 ,并能提高效应细胞的免疫活性 ,起到加强机体免疫力和对抗感染的作用。骨髓抑制是抗癌药物的主要副作用 ,是化学治疗 (简称化疗 )中最显著的障碍因素 ,在临床上表现为粒细胞减少以及与之相关的感染。目前临床已将ｒＨｕＧＭ ＣＳＦ广泛用于强烈化疗及骨髓移植所致的白细胞减少症或粒细胞缺乏症。我们于 2 0 0 0年 3～ 6月将厦门特宝生物工程有限公司研制的ｒＨｕＧＭ ＣＳＦ(商品名 :特尔立 )用于化疗后药物性粒细胞减少症的治疗。对象和…     

哮喘儿粒细胞集落刺激因子和肿瘤坏死因子的变化

目的探讨小儿哮喘血粒细胞集落刺激因子(G-CSF)和肿瘤坏死因子α(TNF-α)变化.方法采用ELISA法测定30例健康儿童、52例哮喘急性期和38例治疗后患儿静脉血G-CSF和TNF-α水平.结果急性期G-CSF阳性38/52例,TNF-α水平高于治疗后和对照组,差异显著(P＜0.01).缓解期25/38例G-CSF转阴,TNF-α与对照组比较仍明显升高(P＜0.01).G-CSF与TNF-α含量呈正相关,急性期r=0.614,缓解期r=0.461 P＜0.01.结论哮喘患儿血清G-CSF和TNF-α增高,参与了哮喘病理炎症发展.     

Treatment of leukemia relapse with recombinant granulocyte-macrophage colony stimulating factor (rhGM-CSF) following unrelated umbilical cord blood transplant: Induction of graft-vs.-leukemia

An infant with congenital leukemia in complete remission (CR1) received an unrelated donor umbilical cord blood cell transplant from a one-HLA disparate donor. The conditioning regimen consisted of thiotepa, busulfan and cyclophosphamide. GVHD prophylaxis consisted of tacrolimus and mini-methotrexate. Engraftment occurred and a bone marrow aspirate obtained on day 28 showed 100% donor cells. The post-transplant course was complicated by skin and liver GVHD, grade III, that responded to therapy with methylprednisolone, anti-thymocyte globulin and daclizumab (Zenapax), in addition to tacrolimus. A bone marrow aspirate obtained on day 187 showed relapse, with 17% blasts. The patient was then treated for 30 days with recombinant human granulocyte-macrophage colony-stimulating factor treatment (rhGM-CSF). A bone marrow aspirate obtained 17 days after the initiation of rhGM-CSF treatment showed 2% blasts. Ascites was the predominant side-effect of the rhGM-CSF treatment. The patient remains in complete remission 24 months after relapse and 30 months after transplantation. This case documents that rhGM-CSF and withdrawal of immunosuppression can induce a durable complete remission after relapse following an unrelated donor cord blood transplant.     

Studies on tissue factor activity and production by leukocytes of human umbilical cord and adult origin.

Human adult and umbilical cord-derived leukocytes were shown to be capable of generating tissue factor activity on exposure to endotoxin and to reduced pH. Blood for leukocyte separation was collected from normal adults and from newly delivered sections of umbilical cord and mixed leukocyte preparations were obtained by separation over methyl cellulose Hypaque. The coagulant activity of the cell suspension was assayed using a one-stage or two-stage method. Cord-derived leukocytes were shown to develop greater coagulant activity than adult-derived leukocytes when stimulated by endotoxin in vitro at 2,000 cells/mm-3. This response to endotoxin was partially inhibited by prior exposure of the cells to prostaglandin (PG) E1 an to L-epinephrine. Acetylcholine stimulated the production of coagulant activity in the absence of endotoxin. Both cord and adult-derived leukocytes (20,000/mm-3) developed coagulant activity when exposed to pH reduction by lactic or hydrochloric acids and this activity was shown to be tissue factor.     

去铁胺联合阿糖胞苷诱导HL-60细胞凋亡作用的实验研究

目的：探讨铁螯合剂联合阿糖胞苷诱导HL-60细胞凋亡的作用。方法：实验分为5组：空白对照组、去铁胺组、阿糖胞苷组、去铁胺+ 阿糖胞苷组、去铁胺+阿糖胞苷+三氯化铁组。上述各组与HL-60细胞共同培养6,12,24,4 8 h。通过测定细胞活力、形态学变化、流式细胞仪检测和DNA凝胶电泳等观察细胞凋亡等方法观察诱导HL-60细胞的作用。结果：去铁胺+阿糖胞苷可协同降低HL-60 细胞活力、抑制HL-60细胞增殖、诱导HL-60细胞凋亡,其作用随培养时间延长及药物浓度增加而增强。结论：铁螯合剂协同阿糖胞苷具有增强诱导HL-60细胞凋亡 的作用。     

单倍体造血干细胞联合第三方脐血移植治疗慢性肉芽肿临床研究

目的探讨单倍体造血干细胞（haplo-HSC）联合第三方脐血（tpCB）移植治疗X连锁慢性肉芽肿（X-CGD）的临床疗效。方法 2014年4月至2018年3月我院收治26例男性X-CGD患儿，采用haplo-HSC联合tpCB移植治疗，移植中位年龄3.5岁，供者25例为父亲，1例为姑姑。HLA配型5/6相合9例，4/6相合12例，3/6相合5例。清髓性预处理方案选用白消安/环磷酰胺/氟达拉滨/抗胸腺球蛋白；预防急性移植物抗宿主病（aGVHD）采用环孢素A和吗替麦考酚酯。于移植第1天分别输注单倍体骨髓造血干细胞和tpCB，移植第2天输注单倍体外周造血干细胞，回输单倍体总有核细胞中位数为14.6×10⁸/kg体重，CD34⁺细胞中位数为5.9×10⁶/kg体重，CD3⁺细胞中位数为2.1×10⁸/kg体重。结果中性粒细胞和血小板植入中位天数分别是移植后12 d和23 d。移植后30 d为供者型完全嵌合，其中25例为单倍体，1例为脐血。无1例发生原发性植入失败和植入功能不良，1例发生继发性植入失败。NADPH氧化酶活力于移植后1个月均恢复正常。Ⅰ~Ⅱ度aGVHD发生率达35%，Ⅲ~Ⅳ度达15%，1例出现慢性GVHD（cGVHD，皮肤局限性），予激素未进展。随访6~52个月，存活25例，其中24例为无病存活（23例不伴cGVHD，1例伴慢性局限型皮肤排异），NADPH氧化酶活性均恢复正常，1例发生继发性植入失败但仍存活；1例于移植后16个月死于病毒性间质性肺炎。5年的EFS率和OS率分别为81%±12%和89%±10%。结论 haplo-HSC联合tpCB是治疗儿童X-CGD的有效方法之一。     

Phenotypic changes of T-lymphocyte subsets induced by interleukin-12 and interleukin-15 in umbilical cord vs. adult peripheral blood mononuclear cells

The decreased incidence of graft-vs.-host disease found following umbilical cord blood (CB) transplantation, and the increased susceptibility of newborns to infections, have been attributed, in part, to functional and phenotypic immaturity of neonatal T cells. We investigated the phenotypic changes of CB T cells induced by two immunoregulary cytokines, interleukin (IL)-12 and IL-15, alone or in combination. Adult peripheral blood (APB) mononuclear cells (MNCs) were also tested for comparison. Prior to culture, the percentages of CD3⁺ CD8⁺, CD3⁺ CD25⁺, and CD3⁺ CD56⁺ cells were significantly lower in CB MNCs than in APB MNCs. IL-15, but not IL-12, significantly increased CD3⁺ CD8⁺ expression among the CB MNCs after 1 week of culture. Combining IL-12 and IL-15, however, resulted in decreased CB CD3⁺ CD8⁺ expression compared with IL-15 alone. The percentage of CD3⁺ CD25⁺ cells in CB MNCs spontaneously increased in the absence of cytokines, while that of CD3⁺ CD56⁺ cells in CB MNCs could not be enhanced with cytokines. In contrast, the percentages of CD3⁺ CD25⁺ and CD3⁺ CD56⁺ cells among the APB MNCs could be increased with IL-12, IL-15, and further with IL-12 and IL-15 combined. Thus, different patterns of T-cell subset changes were demonstrated between CB MNCs and APB MNCs in response to IL-12 and/or IL-15. These data may serve as a foundation for using cytokine therapy in newborns and children receiving CB transplants.     

心肌疾病抗ADP/ATP载体抗体与粒细胞-巨噬细胞集落刺激因子的临床研究

目的探讨病毒性心肌炎（ＶＭＣ）和扩张型心肌病（ＤＣＭ）的自身免疫发病机制。方法采用免疫印转和放射免疫技术检测了３０例ＶＭＣ、１４例ＤＣＭ患儿血浆中抗心肌线粒体ＡＤＰ／ＡＴＰ运载蛋白（ＡＮＴ）抗体和粒细胞－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）。结果ＶＭＣ和ＤＣＭ患儿抗ＡＮＴ抗体和ＧＭ－ＣＳＦ的阳性率分别为７３％和５７％，而２５例正常健康儿分别为０和１２％（Ｐ均＜０．０１），且抗ＡＮＴ抗体与ＧＭ－ＣＳＦ的血浆水平呈正相关（ｒ＝０．４０９１，Ｐ＜０．０５）。结论小儿ＶＭＣ和ＤＣＭ的发病与心肌的自身免疫损伤有关，抗ＡＮＴ抗体和ＧＭ－ＣＳＦ均参与了这一过程。抗ＡＮＴ抗体可作为小儿ＶＭＣ和ＤＣＭ的一种特异性诊断指标。     

Association of tumor necrosis factor, interleukin-6 and cyclooxygenase pathway with lipopolysaccharide-induced intussusception.

INTRODUCTION: Many factors and mechanisms have been proposed as causes for intussusception (IN); however, the etiology remains unclear. Inflammatory mediators such as tumor necrosis factor (TNF) and interleukin-6 (IL-6), which are elevated during infectious diseases, can significantly affect gastrointestinal motility. Motility changes caused by these agents might contribute to the development of IN. The aim of this experimental study was to determine the preventive effects of indomethacin on lipopolysaccharide (LPS)-induced IN in mice and to investigate the role of TNF and IL-6 on intussusception. MATERIALS AND METHODS: Seventy-eight mice were divided into five groups. In the Control group (n=6), no procedure was done. In the Sham group (n=6), 1 ml saline, in the Indomethacin group (n=6), 10 mg/kg of indomethacin, in the LPS group (n=30), 12 mg/kg of LPS was administered intraperitoneally (IP). In the Treatment group (n=30), 10 mg/kg of indomethacin was administered IP following 12 mg/kg of LPS. All animals were laparotomized 6 hours following IP injections. The existence of IN was noted and blood specimens were obtained. TNFalpha and IL-6 plasma level measurements were performed by standard ELISA for mice. The results were compared using the Mann-Whitney U test and one-way ANOVA test. A value of p<0.05 was considered significant. RESULTS: Five mice (1 in the control, 2 in the LPS, 2 in the Treatment group) were excluded from the study. IN was observed in 6 (20%) mice in the LPS group, whereas it was not found in any mice in the Treatment group. Mean TNFalpha and IL-6 levels were statistically higher in the LPS group (394.72+/-403.79; 195.18+/-218.37 pg/ml, respectively) compared to all other groups, including the Treatment group (p<0.05 for each comparison). Within the LPS group of mice, the levels were higher in animals with IN compared to the mice without IN. CONCLUSION: Increased TNFalpha and IL-6 levels induced by LPS correlated well with the occurrence of IN, and a decrease in these levels via cyclooxygenase (COX) inhibition by indomethacin prevented IN from forming in this experimental model.     

Functional and molecular characterization of interleukin-2 transgenic Ewing tumor cells for in vivo immunotherapy

BACKGROUND: Interleukin-2 (IL-2) is a potent cytokine with potential activity against several tumors including Ewing tumors (ET). Side effects of systemic IL-2 can be circumvented by the use of transgenic tumor cells. However, in vitro manipulation may change the overall gene expression profile of tumor cells unfavorably. Therefore, we assessed gene expression profiles, safety, and immunomodulatory efficacy of IL-2 transgenic (IL-2-tg) ET cells in vitro and in NOD/scid mice. PROCEDURE: Viable wild type A673 tumor cells were co-cultured together with irradiated IL-2-tg or mock-transfected cells and HLA matched peripheral blood mononuclear cells. Activation of T and NK cells was assessed by FACS analysis. The effect of irradiated IL-2-tg cells on tumor growth in vivo was investigated by using NOD/scid mice. Gene expression profiles of wild type and transfected cells were analyzed with Affymetrix HG-U95A microarrays. RESULTS: IL-2-tg cells activated and increased the number of T cells and NK cells in vitro. Co-culture with IL-2-tg but not with mock-transfected cells almost completely suppressed wild type tumor cell growth in vitro. Cell depletion experiments indicated a major contribution of NK cells to this tumor cell suppression. Co-transfer of irradiated IL-2-tg cells significantly reduced wild type tumor growth in NOD/scid mice. Side effects in the treated animals were not observed and no tumor growth was observed after injection of irradiated IL-2-tg cells alone. Gene expression profiling revealed a substantial degree of homogeneity of gene expression in transfected and wild type cells and suggests that transfection and selection procedures had no major impact on the gene expression profile. CONCLUSIONS: Next to a high degree of homogeneity between transgenic and wild type cells, our data suggest that irradiated IL-2-tg ET cells can activate cytolytic effector cells. These cells may have therapeutic potential for ET patients.     

Serial changes of serum interleukin-6, interleukin-8, and tumor necrosis factor alpha among patients with Kawasaki disease.

To determine the role of cytokines in Kawasaki disease, serial measurements of serum cytokine levels were done in 60 patients treated solely with aspirin. Coronary artery aneurysms later developed in 12 of them. The results suggest that elevated serum interleukin-6 and interleukin-8 levels during the first week of illness may be associated with a higher risk of coronary aneurysm formation.