CD11b单核苷酸多态性与中国汉族系统性红斑狼疮的研究

目的 分析CD11b基因rs1143679的单核苷酸多态性(SNP)在中国汉族系统性红斑狼疮(SLE)患者中的表达,并阐明该SNP与SLE临床表型的相关性.方法 采用病例对照的研究方法,应用PCR-PFLP以及直接测序技术对中国汉族人群中584例系统性红斑狼疮患者和624例健康对照者进行多态性检测,分析基因型和等位基因频率的分布差异,并与临床表型进行相关性分析.结果 (1)SLE患者中CD11b rs1143679 GA基因型频率为1.89％,大大低于欧美国家的基因型频率,与香港及泰国地区接近.(2) CD11b rs1143679 GA基因型与狼疮肾炎有相关性(P=0.01),而与发病时间、关节炎、血液系统、神经系统损害没有统计学差异(P＞0.05).结论 CD11b rs1143679 SNP与中国汉族人群系统性红斑狼疮易感性有关,并可能参与了狼疮肾炎的发生发展.     

粘附分子CD11a、CD11b、CD62L在恶性淋巴增殖性疾病的表达

目的：观察恶性淋巴增殖性疾病肿瘤细胞表面β２整合素（ＣＤ１１ａ、ＣＤ１１ｂ）及Ｌ－选择素（ＣＤ６２１）的表达变化及其临床意义。方法：用流式细胞仪检测３５例初诊或复发急性淋巴白血病（ＡＬＬ）、４例慢性淋巴细胞白血病（ＣＬＬ）、３０例多发性骨髓瘤（ＭＭ）、１４例淋巴肉瘤白血症及２５例正常人骨髓单个核细胞粘附分子ＣＤ１１ａ、ＣＤ１１ｂ、ＣＤ６２Ｌ的表达。结果：（１）与正常造血细胞比较，ＣＤ１１ｂ、ＣＤ１     

系统性红斑狼疮患者干扰素γ受体基因多态性研究

目的初步探讨干扰素γ受体(interferonγreceptor,IFNγR)的两个氨基酸位点Val14Met和Gln64Arg多态性与系统性红斑狼疮(systemiclupuserythematosus,SLE)的相关性。方法采用聚合酶链反应单链构象多态性和聚合酶链反应限制性片段长度多态性及DNA测序方法对94例SLE患者和80名健康对照者进行基因分型。结果IFNγR1和IFNγR2基因型与SLE易感性无显著相关(P>0.01)。其中Arg64/Arg64基因型在两组患者间的分布差异有统计学意义(P=0.047,OR=2.481,95%可信区间为0.992～6.203)。此基因型在健康对照组中的分布频数高于SLE患者组,可能是一种具有保护作用的基因型。各个基因型的分布情况和优势比显示各基因型的组合与SLE的发病无显著相关(P>0.01)。其中Val14/Val14与Arg64/Arg64基因型的组合在两组患者间的分布差异有统计学意义(P=0.047,OR=2.481,95%可信区间为0.992～6.203)。此基因型组合在健康对照组中的分布频数高于SLE患者组,故这是一种可能具有保护作用的基因型组合。结论IFNγR1和IFNγR2基因型与SLE易感性无显著相关。     

干扰素及其诱导蛋白在系统性红斑狼疮患者中的表达

应用寡核苷酸表达基因芯片技术检测了10例系统性红斑狼疮(SLE)患者外周血白细胞的基因表达谱,与正常对照外周血的基因表达谱作了比较分析。研究中所取血液标本来自符合1982年美国风湿病协会修订的SLE诊断标准的患者共10例(其中包括1个SLE家系中2个SLE患者姐妹) ,男1例,女9例。另选取与患者性别、年龄相匹配的健康人,男5例,女5例,做正常对照。为验证基因芯片结果的可靠性,我们采用TaqMan荧光定量RT PCR方法进行复核分析。我们使用的是MERGEN公司的hu manH0 4包含336 0个点的寡核苷酸基因芯片。在确保基因芯片技术可靠性和重复性的…     

不同复苏溶液对失血性休克大鼠中性粒细胞CD11a、CD11b表达的影响

目的观察不同复苏溶液对失血性休克大鼠PMN表面CD11a、CD11b表达的影响.方法成年Wistar大鼠随机分为0.9%NaCl(NS)、7.5%NaCl(HS)、5%NaCl-3.5%NaAc(HSA)三组,每组6只.动物麻醉后,自股动脉放血,于10min内使MAP下降至5.07-5.47kPa,维持90min.按4mL/kg体重分别注入NS、HS、HSA,5min内输完.随后输注三倍最大失血量的复方氯化钠溶液,40min内输完.分别于休克前、后、给液后1.5h、3h、6h取血0.2mL,流式细胞仪测定PMN表面CD11a、CD11b表达的变化.结果休克后,各组PMN表面CD11a表达下降,但与休克前比较无显著差别.给液后1.5h,各组CD11a表达继续下降;给液后3h,各组CD11a表达稍有回升;给液后6h,各组CD11a表达又呈下降趋势,HSA组CD11a表达显著低于休克前水平;各组间于各时相点均无显著差别.休克后,各组PMN表面CD11b表达增加,与休克前比较有非常显著差别.给液后3h内,各组CD11b表达随观察时间的延长呈进行性增加;给液后6h,NS组CD11b表达继续增加,HS组和HSA组CD11b表达有下降趋势.各组CD11b表达于给液后各时相点均较休克前和休克时非常显著增加,HS组和HSA组CD11b表达于给液后各时相点均低于NS组,但无显著差别.结论液体复苏后,失血性休克大鼠PMN表面CD11a表达呈下降趋势,CD11b表达呈上升趋势,HS和HSA有减弱这种趋势的作用.     

哮喘患者单核细胞CD11c CD14的表达

哮喘是由多种炎性细胞参与的复杂的气道炎症性疾病 ,外周血单核细胞及肺泡巨噬细胞通过递呈抗原给淋巴细胞、分泌多种前炎性和免疫调节因子而与这种炎症反应密切相关。探讨肺泡巨噬细胞与外周血单核细胞结构、功能在哮喘患者中的变化 ,有助于支气管哮喘病理过程的阐明。本研究采用流式细胞仪 ,测定荧光抗体标记单核细胞表面ＣＤ11ｃＣＤ14,以了解哮喘患者单核细胞功能状态及其在哮喘发病中的作用。1　材料与方法1 1　实验对象　哮喘组 :支气管哮喘患者 2 0例 ,其中男 9例 ,女 11例 ,年龄 2 2～ 5 9岁 ,平均 4 0 5岁。均符合 1997年中华医…     

CRP诱导单核细胞表达CD11b和CCR2并影响脂蛋白对CD11b和CCR2表达的观察

目的探讨C反应蛋白(CRP)对单核细胞表达CC趋化因子受体2(CCR2)和CD11b的作用以及与脂蛋白之间的相互作用。方法以不同浓度的CRP和／或脂蛋白处理THP-1单核细胞,应用流式细胞仪检测细胞表面CCR2、CD11b蛋白的表达,并应用半定量RT-PCR方法检测CCR2的mRNA表达,同时检测处理后的单核细胞与内皮细胞的黏附率以及培养液中NO含量。结果CRP以剂量依赖性方式增加单核细胞表达CCR2和CD11b,RT-PCR结果显示CRP在转录水平上诱导CCR2的表达。不同脂蛋白对CCR2和CD11b的表达作用不同:氧化的低密度脂蛋白(OX-LDL)诱导CCR2和CD11b的表达(P<0．01),而高密度脂蛋白(HDL)抑制此二者的表达(P<0．01),天然低密度脂蛋白(LDL)则对其无显著影响(P<0．01)。CRP抑制OX-LDL所诱导的CCR2(115．7±6．40比99．0±3．65, P<0．01)和CD11b(121．3±4．79比98．5±4．90,P<0．01)的表达增加;但与LDL显著地协同上调CCR2(LDL 50μg/ml比LDL 50μg/ml+CRP 10μg／ml;CRP 10μg/ml比LDL 50μg/ml+CRP 10μg/ml,均P <0．01)和CD11b(LDL 50μg/ml比LDL 50μg/ml+CRP 10μg／ml;CRP 10μg／ml比LDL 50μg/ml+CRP 10μg/ml,均P<0．01)的表达;CRP削弱HDL对CCR2和CD11b表达的抑制作用。培养液中的NO含量与CCR2和CD11b密切相关。结论CRP诱导单核细胞表达CCR2和CD11b,并调节脂蛋白对CCR2和CD11b的作用;NO可能是此过程中的信使之一。     

SLE患者TNF-β基因多态性分析

系统性红斑狼疮（SEE）是一种累及多系统、具有多种自身抗体的自身免疫性疾病,遗传因素在SLE发病机制中起重要作用。肿瘤坏死因子（TNn是具有多种生物学功能的细胞因子,体内外实验证实,TNF水平与SLE的活性指标密切相关。鉴于TNF基因的特殊位置及TNF的重要生物学活性,TNF的基因多态性与SLE的发病、病程及预后之间的关系已引起普遍关注。我们早期研究发现．TNF—α基因多态性与SLE发病相关,为探讨TNF-β基因的多态性与SLE的关系．     

安徽省汉族SLE患者和正常人RANTES单核苷酸多态性及CCR5基因多态性的比较

目的　探讨中国汉族系统性红斑狼疮 (SLE)患者和正常人群RANTES单核苷酸多态性(SNP)及其受体CCR5多态性。方法　收集 14 6例确诊的SLE患者和 15 9名正常对照。通过PCR RFLP方法检测研究对象RANTES启动区SNP及其受体CCR5△ 32突变频率。结果　病例组RANTES 4 0 3位点G G、G A、A A基因型频率分别为 76 .71%、2 1.92 %、1.37% ;对照组分别为 6 7.30 %、2 9.5 6 %和3.14 % ,两组间基因型分布差异无显著性 (P >0 .0 5 )。两组突变等位基因 4 0 3A频率分别为 12 .3%、17.9% (P >0 .0 5 )。病例组RANTES 2 8位点C C、C G、G G基因型频率分别为 93.15 %、6 .85 %、0 ;对照组分别为 86 .79%、12 .5 8%和 0 .6 3% (P >0 .0 5 )。两组突变等位基因 2 8G频率分别为 3.4 %、6 .9% (P>0 .0 5 )。病例组和对照组突变等位基因CCR5△ 32频率分别为 0、0 .3% (P >0 .0 5 )。中国汉族人群RANTES突变基因型 4 0 3A A低于北美黑人和西非黑人 (P <0 .0 5 ) ,与北美高加索、北美西班牙人和北美亚洲人一致。RANTES 2 8位点基因型分布与北美亚洲人一致 ,但与北美高加索、北美西班牙人、北美黑人和西非黑人相差较大 (P <0 .0 5 )。结论　本次研究发现RANTES 4 0 3位点和 2 8位点单核苷酸多态性与SLE发病没有直接的关系。CCR5△ 32     

SLE模型小鼠高IgG血症易感基因-Fcgr2b基因突变及对其表达的影响

目的:测定系统性红斑狼疮(SLE)小鼠模型 (NZB×NZW)F1双亲NZB ,NZW小鼠Fcgr2b基因启动子区核酸序列及其表达的改变,明确Fcgr2b基因的突变性质、对该基因表达的影响及与高IgG血症的关系。方法:采用核酸序列分析测定NZB ,NZW小鼠Fcgr2b基因启动子区突变性质;RT PCR检测Fcgr2b基因表达的改变;ELISA法测定比较(NZB×NZW)F1,NZB和NZW小鼠及(NZB×NZW)F1×NZW回交小鼠Fcgr2b基因B/W型与W /W型组间血清总IgG水平。结果:NZB小鼠Fcgr2b基因启动子区与正常鼠BALB/c相比存在2个部位碱基缺失,分别为13bp及3bp。NZW小鼠除有3个碱基置换外,与BALB/c鼠该基因启动子区序列相同;NZB小鼠Fcgr2b基因mRNA的表达较正常鼠BALB/c降低;NZB小鼠血清总IgG水平明显高于NZW小鼠(P <0 0 5 ) ,回交小鼠Fcgr2b基因B/W型组血清总IgG水平明显高于W/W型组(P <0 0 0 0 1)。结论:NZB小鼠Fcgr2b基因启动子区存在碱基缺失,且该缺失突变可引起其表达的降低而导致高IgG血症     

Increased CD11b expression on polymorphonuclear leucocytes and cytokine profiles in patients with Kawasaki disease

Clinical evidence implicates polymorphonuclear leucocytes in the pathogenesis of vasculitis in Kawasaki disease. We examined modulation of expression of adhesion molecules (CD11b and CD62L) on polymorphonuclear leucocytes and how this expression is related to serum cytokine concentrations. In 18 patients with Kawasaki disease and 15 control subjects, adhesion molecule expression was determined by two-colour immunofluorescence staining of blood leucocytes and flow cytometry. Eight cytokines and chemokines were also measured. In patients with Kawasaki disease, mean fluorescence intensity for CD11b before giving intravenous immunoglobulin was significantly higher than in normal subjects (P<0 x 005). After intravenous immunoglobulin, mean fluorescence intensity for CD11b decreased significantly. With coronary artery lesions present, mean CD11b fluorescence intensity was significantly higher than without coronary artery lesions (P=0 x 005 before intravenous immunoglobulin; P=0 x 024 after intravenous immunoglobulin). No differences were seen in CD62L expression on polymorphonuclear leucocytes between patients with Kawasaki disease and normal subjects. CD11b expression on polymorphonuclear leucocytes correlated positively with serum interleukin (IL)-6, IL-10, granulocyte colony-stimulating factor, percentage of neutrophils among white cells and C-reactive protein. Polymorphonuclear leucocytes from patients with Kawasaki disease showed increased CD11b expression, which was associated with increased serum cytokines and appeared to be related to coronary artery lesions.     

Peripheral blood phagocyte CD14 and CD11b expression on admission to hospital in relation to mortality among patients with community-acquired infection

Objective and Design: Prognostic value of markers of systemic inflammation were evaluated in patients admitted to hospital. Material: The study comprises 327 patients with community- acquired infection verified on admission (n = 290) or within 3-day follow-up (n = 37). Methods: On-admission levels of phagocyte CD11b/CD18 and CD14 expression were measured using whole blood flow cytometry. Clinical data were collected retrospectively from medical records. Results: In univariate analysis, non-survivors as compared to survivors had higher age, lower arterial pressure, higher heart rate, and lower monocyte CD14 density. In multivariate analysis high age [relative mortality RR 1.05 (95% CI 1.01 to 1.08), p = 0.016] and low CD 14 expression on monocytes [RR 7.49 (CI 1.63 to 34.33), p = 0.01] remained predictive for the 28-day mortality. Conclusion: In patients with community-acquired infection, low on-admission level of monocyte CD14 is related to fatal outcome. Received 4 May 2005; returned for revision 4 June 2005; accepted by K. Visvanathan 9 July 2005     

Total and biologically active CD154 in patients with SLE

CD154, a member of the tumor necrosis factor receptor family, is involved in several biological responses. In the sera of systemic lupus erythematosus (SLE) patients, the levels of sCD154 have been shown to be increased, however, few reports have dealt with the biologically active tetramer. Here, we assessed the biological activity of the serum CD154 tetramer using bioassays for BC activation and production nitrite or peroxide. The patients showed a markedly increased total sCD154 serum concentration (12.5 ± 8.2 vs. 3.9 ± 1.2 ng/ml; p < 0.001). ba-sCD154 was significantly increased in non-treated patients (7.4 ± 3.4 ng/ml, n = 22; p < 0.001) and patients with the highest SLE disease activity index (SLEDAI) scores (5.3 ± 2.9 ng/ml, n = 8), but not in stable patients (1.3 ± 1.2 ng/ml, n = 30) whose values were similar to normal healthy donors (NHD; 0.8 ± 0.2 ng/ml). Patients with SLEDAI above 8 that recovered after successful treatment displayed significantly decreased levels of ba-sCD154. We conclude that the bioassay is a useful tool discriminating active and stable SLE, as well as non-treated patients.     

c-kit gene mutation and CD117 expression in human anorectal melanomas

c-kit and BRAF mutations play an important role during the pathogenesis of melanoma. The subtypes of melanomas arising from different parts of the body have variable c-kit or BRAF mutation frequencies. Few studies in the literature have examined c-kit and BRAF mutation status in melanomas that occur in the anus and rectum. In this study, we analyzed 40 cases of anorectal melanoma for c-kit and BRAF mutations by DNA sequencing using paraffin-embedded tissues. c-kit Mutations were detected in exons 9, 11, 13, and 17. CD117 expression in tumor cells was analyzed by immunohistochemistry. Our study showed that a c-kit mutation was found in 7 of the 40 cases of anorectal melanoma. CD117 expression was detected in 16 of the 40 cases, and 3 of these 16 cases also had c-kit mutations. Mutations in BRAF were also identified in 2 patients. These results indicate that a subset of anorectal melanomas have activating c-kit mutations, which suggests that kinase inhibitors such as imatinib may be used to treat this subset of melanoma patients. In addition, our results show that c-kit mutations do not correlate with CD117 expression.     

In vivo transmigrated monocytes from patients with stable coronary artery disease have a reduced expression of CD11b

Coronary artery disease (CAD) is characterized by infiltration of monocyte derived cells in the intima of the vessel wall. We hypothesized that accumulation of these cells is caused partly by an altered monocyte transmigration process in CAD. To gain insight into this issue we applied the skin blister method that allows collection of in vivo transmigrated cells at sites of local inflammation. Nineteen patients with stable CAD and 19 matched controls were enrolled. Markers of inflammation and gradients of chemokines, as well as adhesion molecule expression and up-regulation capacity, were studied. The expression of inflammatory markers, such as C-reactive protein, interleukin (IL)-6, tumour necrosis factor-alpha and IL-10, was similar in patients and controls, indicating that patients were in a stable phase of the disease. Expression of adhesion molecules, CD11b and very late activation antigen-4, on peripheral monocytes did not differ between patients and controls. However, following in vivo transmigration, monocytes in patients with CAD had a significantly reduced expression and mobilization of CD11b. The effect on CD11b could not be reproduced by in vitro stimulation with blister fluid, representing a local inflammatory milieu, or in an in vitro system of transmigration. These findings point towards differences in monocyte CD11b expression and availability at an inflammatory site between patients with CAD and healthy controls.     

Human granulocyte CD11b expression as a pharmacodynamic biomarker of inflammation

A method has been developed for the direct quantification of the CD11b integrin on granulocytes by flow cytometric analysis of whole blood specimens following either LTB₄ or lipopolysaccharide (LPS) stimulation. This method has utility in evaluating the pharmacodynamic action of either LTB₄ receptor antagonists or immune cell modulators in effecting CD11b integrin expression and granulocyte activation in human subjects administered such drugs. Previous studies using CD11b as a biomarker of granulocyte activation have faltered because of the difficulty in controlling the activation state of the granulocyte following removal of blood from subjects. The present study has made use of a newly validated method using either LTB₄ or LPS to stimulate CD11b expression on granulocytes and has been used, as one measure, in the evaluation of LPS activity when administered to normal human volunteers.     

Effect of cysteinyl leukotriene receptor antagonist on CD11b and CD23 expression in asthmatic children

BACKGROUND: Cysteinyl leukotrienes are potent pro-inflammatory mediators that contribute to the pathophysiologic features observed in allergic asthma. Inhibitors of leukotriene receptors represent novel therapy in asthma treatment. In addition to the protection from early asthmatic responses, these drugs have recently been shown to protect from late airway responses too. METHODS: We studied the effect of treatment with an oral antagonist of cysteinyl leukotriene receptors on the increased expression of the low-affinity IgE receptor, CD23, on B cells, and of its ligands, CD11b and CD11c, on CD4(+) T cells and monocytes in peripheral blood of patients with allergic asthma. In this uncontrolled open-label study, 14 children with allergic asthma received montelukast, a cysteinyl leukotrine receptor antagonist, for a period of 6 weeks after demonstrating forced expiratory volume in 1 s (FEV(1)) of less than 80% of the predicted value. Samples of peripheral heparinized blood and sera were obtained before and after therapy completion. Three-colour immunofluorescence analysis was performed, and expression of CD11b and CD11c on CD4(+) T lymphocytes and monocytes as well as the expression of CD21 and CD23 on B cells were determined (n=12). Peripheral blood eosinophil count, changes in FEV(1) and peak expiratory flow rate (PEFR), asthma exacerbations, and as-needed use of beta-agonist were also monitored. RESULTS: Montelukast improved FEV(1) and PEFR, and decreased peripheral eosinophil counts in all study patients. There was no significant change in the expression of CD21 and CD23 on B cells. The expression of CD11c on CD4(+) T cells and of both CD11b and CD11c on monocytes remained similar to the pretreatment expression. However, the percentage of CD11b(+)CD4(+) T lymphocytes significantly decreased after treatment with montelukast. This was accompanied by a significant decrease in the levels of total IgE. CONCLUSION: The capacity of 6-week montelukast therapy to reduce the percentage of CD11b CD4(+) T cells might be a mechanism leading to the immune response modulation on this T cell subset interaction with CD23-expressing B cells and subsequent down-regulation of IgE synthesis.     

A novel mutation in STK11 gene is associated with Peutz-Jeghers Syndrome in Indian patients

Background  

Peutz-Jeghers syndrome (PJS) is a rare multi-organ cancer syndrome and understanding its genetic basis may help comprehend the molecular mechanism of familial cancer. A number of germ line mutations in the STK11 gene, encoding a serine threonine kinase have been reported in these patients. However, STK11 mutations do not explain all PJS cases. An earlier study reported absence of STK11 mutations in two Indian families and suggested another potential locus on 19q13.4 in one of them.