脐血CD34~+细胞体外短期培养扩增研究

为寻找更有效的体外扩增脐血CD34 + 细胞的造血细胞因子组合 ,采集健康产妇脐带血 ,用免疫磁珠法分选CD34 + 细胞。采用SCF、FLT3 L、TPO和IL 34种具有早期作用的细胞因子的不同组合进行脐血CD34 + 细胞短期无血清液体培养 ,观察培养前后有核细胞、CD34 + 细胞、CD34 + /CD38- 细胞、CFU GEMM、CFU GM和BFU E数量的变化。结果在 3种不同的细胞因子组合中 ,同时应用SCF、FLT3 L、TPO和IL 34种细胞因子培养 7d的扩增效果最好。突出的发现是在这种条件下CD34 + /CD38- 细胞亚群达到平均 1 97.9倍的扩增效果。提示 :SCF、FLT3 L、TPO和IL 34种细胞因子是脐血CD34 + 细胞体外扩增理想的细胞因子组合     

大肠黏膜癌变过程中PTEN和p27的表达及相关性研究

目的 :探讨PTEN及p2 7在大肠黏膜癌变过程中的表达及两者的相关性。方法 :采用免疫组织化学S -P法检测了 5 8例大肠癌 ,15例腺瘤性息肉及 11例配对的癌旁正常组织中抑癌基因PTEN和 p2 7的蛋白表达。分析PTEN和 p2 7的表达情况及与各种临床病理特征的关系及两者的相关性。结果 :在癌旁正常组织、大肠腺瘤性息肉、大肠癌组织中PTEN表达率分别为 (97.3± 4 .3) % ,(85 .2± 16 .8) % ,(13.8± 17.6 ) % ,呈递减趋势。大肠癌DukesA/B期组PTEN蛋白的高表达率为 88.9% ,明显高于DukesC/D期组中的 5 1.6 %。PTEN的高表达率与性别、年龄、肿瘤的大小、部位、分化程度、有无淋巴结转移无关。p2 7主要表达在细胞核 ,也有少量表达在细胞浆。在癌旁正常组织、大肠腺瘤性息肉、大肠癌组织中p2 7高表达率分别为 (98.8± 1.0 8) % ,(86 .0± 13.6 ) % ,(5 6 .8± 2 6 .0 ) % ,呈递减趋势。在大肠癌高、中分化组中 p2 7蛋白的表达率为 71.4 % ,明显高于低分化组中的 2 3.1%。p2 7的高表达率与患者的Dukes分期、性别、年龄、肿瘤的大小、部位及有无淋巴结转移无关。在大肠癌组织中 ,PTEN与 p2 7蛋白的表达呈正相关 (r =0 .6 4 2 )。结论 :PTEN及p2 7表达在癌旁正常组织、大肠腺瘤性息肉、大肠癌组织中均呈递减趋势 ,提示P     

Lectin-induced increase in clonogenic growth of haematopoietic progenitor cells

Abstract: The galactoside-specific plant lectin, Viscum album agglutinin (VAA-I) increases cellular parameters of natural host defence. It also binds to a variety of haematopoietic cells, including progenitors. We investigated whether VAA-I has a stimulatory effect on haematopoietic progenitor cells. Peripheral blood progenitor cells from 7 healthy volunteers were cultured in a colony assay with VAA-I plus erythropoietin (EPO) and stem cell factor (SCF). At 50 pg/ml VAA-I induced a significant increase in the cytokine-dependent clonogenic growth (52% in median, p<0.05). In another set of experiments purified CD34+ cells were isolated from the bone marrow aspirate of 4 patients with non-metastatic breast cancer using fluorescence-activated cell sorting. Binding to CD34+ cells was demonstrated by using directly fluorescence-conjugated VAA-I. Co-incubation with d -galactose significantly abrogated this effect. CD34+ cells were cultured in the presence of EPO, SCF, interleukin-3, granulocyte/monocyte colony-stimulating factor and granulocyte colony-stimulating factor. VAA-I alone had no measurable effect on the clonogenic growth of the isolated cells. However, at concentrations of 100 and 250 pg/ml VAA-I increased the cytokine-dependent proliferation and differentiation of CD34+ cells by a median of 75 and 85%, respectively. The results show that VAA-I binds to haematopoietic progenitor cells and has a co-stimulatory effect on their proliferation.     

Interactions between peripheral blood CD8 T lymphocytes and intestinal epithelial cells (iEC)

Intestinal intraepithelial lymphocytes (iIEL) are primarily CD8 cells and most of them have a CD28^? phenotype, the phenotype of effector cytotoxic T cells. We asked whether the predominance of CD8⁺ CD28^? T cells in the gut may result from peripheral blood T cells preferentially migrating to the iIEL compartment and adhering to iEC. Compared with CD4 cells, adhesion of resting CD8⁺ T cells to iEC cell lines was significantly higher. Adhesion could be blocked with a MoAb to gp180, a molecule expressed on iEC which is known to interact with CD8/lck. No significant difference in the level of adhesion was observed between CD8⁺ CD28⁺ and CD8⁺ CD28^? T cells. Thus CD8 cells may preferentially migrate to the iIEL compartment, but loss of CD28 expression could occur in situ after migration. Consistent with this hypothesis, the CD8⁺ CD28^? cells became enriched after co-culturing T cells with iEC cell lines and primary iEC. Induction of the CD8⁺ CD28^? phenotype in cord blood and adult T cells was observed in co-cultures with iEC and also with mitogens and superantigens. In the latter case, CD28 down-modulation was seen specifically in the Vβ subset targeted by the superantigen, indicating that loss of CD28 expression is a direct result of T cell receptor (TCR)-mediated stimulation. The combined results suggest that CD8⁺ CD28^? T cells are antigen experienced T cells, and that they may have a survival advantage in the presence of gut epithelial cells in vitro. This may contribute to the predominance of CD8⁺ CD28^? T cells in the iIEL compartment.     

Overexpression of suppressor of cytokine signalling-5 augments eosinophilic airway inflammation in mice

BACKGROUND: Enhanced expression of the suppressor of cytokine signalling (SOCS)-5 might be of therapeutic benefit for T-helper type 2 (Th2) dominant diseases, as its expression is reported to result in a reduction of Th2 differentiation in vitro due to the inhibition of IL-4 signalling. OBJECTIVE: To investigate the regulatory role of SOCS-5 in vivo, we explored the phenotype of an experimental asthma model developed in SOCS-5 transgenic (Tg) mice. METHODS: The SOCS-5 Tg mice or wild-type (WT) mice were sensitized and repeatedly challenged with ovalbumin (OVA). We examined bronchoalveolar lavage fluid (BALF), lung specimens, and airway hyperresponsiveness (AHR) to methacholine. RESULTS: The production of IFN-gamma by CD4(+) T cells from unprimed SOCS-5 Tg mice was significantly increased in comparison with unprimed wild-type mice, indicating that SOCS-5 Tg mice have a Th1-polarizing condition under natural conditions. However, in an asthma model, significantly more eosinophils in the airways and higher levels of IL-5 and IL-13 in BALF were observed in the SOCS-5 Tg than the wild-type mice. AHR in the asthma model of SOCS-5 Tg was also more enhanced than that of wild-type mice. OVA-stimulated CD4(+) T cells from the primed SOCS-5 Tg mice produced significantly more IL-5 and IL-13 than CD4(+) T cells from wild-type mice. CONCLUSION: Our results demonstrate that the overexpression of SOCS-5 does not inhibit Th2 response, but rather augments the phenotype of the asthma model in vivo. This finding throws into question the therapeutic utility of using enhancement of SOCS-5 expression for Th2-dominant disease.     

HLA-B_(27)与急性前葡萄膜炎相关性的临床研究

目的 :应用SSP PCR(Sequencespecialprime polymerasechainreaction)基因检定技术对急性前葡萄膜炎 (a cuteanterioruveitis ,AAU)患者HLA B2 7基因进行检测 ,并且对HLA B2 7阳性与阴性患者临床特征加以分析。方法 :采用SSP PCR基因检定技术检测 98例AAU患者及 82例正常人样本的HLA B2 7基因。并对HLA B2 7阳性与阴性患者临床特征进行观察。结果 :98例AAU患者样本中有 5 7例样本呈HLA B2 7阳性 ,82例正常人样本有 4例样本呈HLA B2 7阳性 ,阳性率分别为 5 8.2 %和 4 .9%。经 χ2 检验 ,χ2 =4 1.33,P <0 .0 0 5 ,二组间有显著差异。HLA B2 7阳性患者多见于男性 ,单眼多见 ,粉尘状KP ,发病时视力下降明显 ,易于复发 ,且并发症少为其特征 ,激素治疗效果佳。结论 :采用SSP PCR基因检定技术测定HLA B2 7快速、简单、准确性高、客观性强 ,值得推广和应用。HLA B2 7与急性前葡萄膜炎有着高度相关性。HLA B2 7阳性患者与阴性患者在临床特征上有着一定程度的差异。     

胃癌根治术病人围术期异体输血后外周血单核细胞CD40L表达的变化

目的 研究胃癌根治术病人围手术期异体输血外周血单核细胞(PBMCs)白细胞分化抗原40配体(CD40L)表达的变化。方法 胃癌根治术病人30例,随机分为3组,每组10例。A组围术期不输血,B组围术期输入去白细胞的全血,C组围术期输入异体全血。另选10例健康人作为对照。分别在手术前、术后2、5、10 d采外周静脉血5 ml,用Ficoll分离液梯度离心法分离出PBMCs和血浆,将PBMCs置于自身血浆环境中,并在植物血凝素(PHA,20 mg/L)的刺激下进行培养,48 h后收获细胞,用流式细胞术检测CD40L表达。结果 健康人外周血未受PHA刺激时检测不到CD40L的表达,经PHA刺激后CD40L 细胞占CD4 T细胞的百分数为1.7％±0.4％,与三组胃癌病人术前比较差异无显著性(P>0.05)。与术前比较,B组术后2 d PBMCs CD40L表达升高(P<0.05),C组术后各时点升高(P<0.05);与A组比较,B组术后2 d升高(P<0.05),C组术后各时点升高(P<0.05);与B组比较,C组术后各时点升高(P<0.05)。结论 围手术期异体输血可造成免疫抑制,输异体血后CD40L表达增加,且输全血比输去白细胞的全血更明显。围手术期成分输血优于输注全血。     

Rosiglitazone down-regulates lipopolysaccharide-induced expression of CD40 and intercellular adhesion molecule 1 in rat peritoneal mesothelial cells through a NF-κB dependent mechanism

Objective To investigate the effect and mechanism by which PPARγ ligand, rosiglitasone, regulates the expression of CD40 and intercellular adhesion molecule 1 (ICAM-1) in the rat peritoneal mesothelial cells (RPMCs) induced by lipopolysaccharide (LPS). Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and maintained under defined in vitro conditions. The cells were randomly divided into groups as follows: medium, LPS (5 mg/L), LPS (5 mg/L)+BAY11-7085(5 μmol/L, NF-κB inhibitor), rosiglitazone (10 μmol/L or 20 μmol/L, peroxisome proliferator-activated receptor γ activator), LPS (5 mg/L)+rosiglitazone (10 μmol/L)+GW9662 (3 μmol/L, peroxisome proliferator-aetivatcd receptor γ antagonist), and LPS (5 mg/L)+vehicle (DMSO 0.2 ml/L). The expressions of CD40 and ICAM-1 RNA in RPMCs were examined by RT-PCR after 3 hour treatment, and the protein expressions of CD40, ICAM-1, p-NF-κB p65 and p-IκBα were examined by Western blot or immunofluorescence after 24 hour treatment. Results Following treatment with LPS, both the expressions of CD40 and ICAM-1 protein in RPMCs were up-regulated significantly (P<0.05), and the phosphoralation of p65 was increased greatly (1.10±0.17 vs 0.55±0.06, P<0.05). BAY11-7085 (5 μmol/L) significantly decreased the protein expression of p-p65 (0.22±0.11 vs 1.10±0.17, P<0.01), CD40 (0.34±0.02 vs 0.50±0.06, P<0.05) and ICAM-1 (0.35±0.16 vs 0.74±0.03, P<0.05). Pretreated with rosiglitazone for 3 h then added with LPS for 1 h, the levels of p-p65, CD40 and ICAM-1 in RPMCs were significantly decreased compared with those of LPS group (0.77±0.08 vs 0.90±0.10, P相似文献   

CD11b在肺纤维化患者的表达及红霉素对其影响

目的探讨CD11b在肺纤维化患者外周血中性粒细胞上的表达及红霉素对它的影响.方法肺纤维化患者(A组,n=13)13例经红霉素治疗3个月,收集治疗前后及健康对照组(B组,n=13)13例的外周血中性粒细胞涂片,用桥联酶免疫法测定CD11b表达阳性的中性粒细胞百分率.结果A组治疗前后,患者外周血中性粒细胞CD11b的表达阳性率均显著高于健康对照组(P＜0.01);A组治疗前,患者外周血中性粒细胞CD11b的表达阳性率显著高于治疗后(P＜0.01).结论肺纤维化患者外周血中性粒细胞CD11b的表达阳性率显著高于健康对照组;红霉素能抑制肺纤维化患者外用血中性粒细胞CD11b的表达.