系统性红斑狼疮外周血单个核细胞CD28/B7分子mRNA的表达

目的 :探讨CD2 8 B7分子在系统性红斑狼疮 (SLE)发病机制中的作用及其临床意义。方法 :应用逆转录 聚合酶链反应 (RT PCR)检测 35例活动期SLE患者和 30例正常人外周血单个核细胞 (PBMC)中CD2 8、B7 1和B7 2mRNA的表达水平。结果 :35例活动期SLE患者PBMC中CD2 8的阳性表达率 (2 2 86 % )明显低于正常人对照组 (70 0 0 % ) ,差异非常显著 (P <0 0 0 1) ;B7 2的阳性表达率 (82 86 % )明显高于正常对照组 (5 3 33% ) ,差异显著 (P <0 0 1) ;活动期SLE组CD2 8的平均表达水平 (0 194 5± 0 2 0 74 )明显低于正常对照组 (0 4 2 38± 0 10 5 3) ,差异显著 (P <0 0 5 ) ;B7 2的平均表达水平 (0 86 75± 0 2 5 75 )明显高于正常人对照组 (0 4 898± 0 30 72 ) ,差异非常显著 (P <0 0 1) ;35例活动期SLE患者中仅有 2例B7 1呈阳性表达。结论 :CD2 8 B7分子的异常表达可能与SLE患者淋巴细胞和抗原呈递细胞 (APC)的功能变化有关。B7 1低水平与B7 2的高水平表达表明 ,SLE患者T细胞的活化可能主要是通过CD2 8与B7 2的交联传递共刺激信号 ,介导以Th2型反应为主的免疫应答反应 ;B7 2的表达水平可能与SLE疾病的活动性有一定的相关性。CD2 8mRNA的低水平表达可能与外周血CD2 8 T细胞凋亡增加或迁移到炎症部     

系统性红斑狼疮患者淋巴细胞共刺激分子异常表达的研究

目的 探讨系统性红斑狼疮(SLE)患者淋巴细胞共刺激分子的表达及意义.方法 采用流式细胞术检测SLE患者淋巴细胞CD28、CTLA-4、CD80和CD86的表达,并与对照组比较.结果 与健康对照者相比较,SLE患者CD3+细胞增加(P<0.05),CD3+CD4+细胞降低(P<0.05),CD3+CD8+细胞升高(P<0.05),CD4/CD8比例明显倒置(P<0.01),CD28表达降低,CTLA-4升高,CD86的表达显著高于正常人(P<0.01),CD80在SLE患者CD19+B细胞上的表达与正常人无异(P>0.05).结论 T细胞亚群改变及T、B淋巴细胞共刺激分子CD28、CTLA-4和CD86在SLE发病机制中起重要作用.     

CD28受体分子的研究进展

人ＣＤ２８分子为共刺激受体分子家族中的主要成员之一，是目前国内外研究的一个热门课题，通过激活或抑制ＣＤ２８分子独特的信号传导途径，探讨机体的免疫调节功能，可揭示Ｔ细胞活化的机理。人Ｄ２８分子为同源二聚体糖蛋白，Ｍｒ４４０００，ＯＲＦ编码２２０个氨基酸，主要表达于Ｔ细胞表面，在Ｔ细胞的活化，增殖，淋巴分子的产生，以及免疫调节等方面发挥着重要的作用，本文仅对人ＣＤ２８分子受体的家族、生物学功能与机理等     

系统性红斑狼疮患者外周血淋巴细胞CD1c的表达

目的 :研究系统性红斑狼疮 (SLE)患者外周血淋血细胞CD1c的表达情况及与疾病活动性之间的关系。方法 :用流式细胞仪检测了 4 7例SLE患者外周血淋巴细胞CD1c的表达及淋巴细胞表型分析 ,并评价与疾病活动性的关系。结果 :SLE活动组病人CD1c 细胞百分率显著增高 (P <0 0 5 ) ,CD4 细胞百分率显著降低 (P <0 0 1) ,CD3 、CD8 细胞百分率正常 ,CD2 0 细胞数增高 (P <0 0 1)。稳定期病人CD1c 细胞百分率正常 ,CD4 、CD8 、CD2 0 细胞百分率均正常。SLE患者CD1c细胞阳性率与患者SLEDAI的评分有显著的相关性 (r=0 6 8,P <0 0 1) ,与抗dsDNA抗体的表达有显著相关性 (r =0 36 ,P <0 0 5 ) ,与抗心磷脂抗体的表达有显著的相关性 (r=0 6 4 ,P <0 0 1) ,与血清C3水平有显著相关性 (r =- 0 35 ,P <0 0 5 )。活动期病人经治疗后CD1c的表达明显下降。结论 :系统性红斑狼疮患者外周血CD1c表达与疾病的活动性明显相关 ,CD1c可能在SLE脂类抗原及核酸类抗原的递呈及抗双链DNA抗体、抗磷脂抗体的产生中起重要作用。     

CD4+CD25+T细胞在系统性红斑狼疮中的表达及意义

目的：探讨CD4^＋ CD25^＋ T细胞、IL-10在系统性红斑狼疮（SLE）患者外周血的表达及临床意义。方法：入组30例SLE患者和20例正常对照者，其中活动性SLE患者17人，非活动性SLE患者13人。用流式细胞仪检测SLE患者和正常对照者的外周血CD4^＋ CD25^＋ T细胞阳性率，用酶联免疫吸附试验（ELISA）检测血清中IL-10浓度。结果：活动性和非活动性SLE患者CD4^＋ T细胞总数均低于正常对照者；活动性和非活动性SLE患者CD4^＋ CD25^＋ T细胞阳性率高于正常对照者；活动性SLE患者IL-10浓度显著高于非活动性SLE患者和正常对照者。SLE患者CD4^＋ CD25^＋ T细胞阳性率和血清IL-10浓度与补体C3、抗DNA抗体水平及SLEDAI积分均无相关性。结论：SLE患者外周血CD4^＋ CD25^＋ T细胞是活化T细胞的标志，IL-10分泌异常与SLE的发病有关。     

共刺激分子CD28：CTLA4／B7及CD40／CD40L和SLE

共刺激分子CD28：TCLA-4／B7和CD40／CD40L对B细胞的增殖、分化、抗体的分泌和T细胞的活化、细胞因子的分泌具有重要作用，其对系统性红斑狼疮的发生、发展具有重要的促进作用。本文综述了CD28：TCLA-4／B7和CD40／CD40L生物学特性、功能及在系统性红斑狼疮发生、发展中的作用，以及免疫学治疗的临床应用前景。     

重症肌无力外周血单个核细胞CD28-B7共刺激通路相关分子的表达

目的 :探讨重症肌无力 (MG)患者CD2 8 B7共刺激信号通路相关分子的表达及其与MG发病的关系。方法 :用流式细胞仪检测 18例MG患者和 16例健康对照者外周血CD2 8、CTLA4、B7 1、B7 2分子在CD4 T细胞和CD8 T细胞表面的表达。结果 :MG组外周血CD2 8、CTLA4、B7 1、B7 2分子的表达均增强 ,其中CD2 8 细胞的增多主要表现在CD4 T细胞亚群 ,CT LA4的表达增强主要在CD8 T细胞 ,B7 1和B7 2分子在CD4 或CD8 T细胞的表达 ,MG组与对照组无差别。结论 :MG的发病与CD2 8共刺激通路的活化密切相关 ,检测外周血CD2 8共刺激相关分子的表达可反映MG患者的免疫反应状态 ,有助于临床诊断和用药。     

CD226在系统性红斑狼疮患者T淋巴细胞亚群上的表达

目的：研究CD226在SLE患者外周血T淋巴细胞亚群上的表达，以阐明CD226抗原在SLE患者体内T细胞活化中作用以及与SLE发病的关系。方法：31例SLE患者和30例健康志愿者外周血单个核细胞，在体外培养72h后，三色荧光标记的单克隆抗体染色，利用流式细胞仪测定T细胞亚群细胞表面CD226抗原的表达。结果；SLE患者组的CD3^ 、CD4^ 、CD8^ T淋巴细胞上CD226^ 表达均高于正常对照组（P＜0．01）；活动期SLE组、静止期SLE组的CD3^ 、CD4^ 、CD8^ T细胞上CD226^ 表达均显著高于对照组（P＜0．01），而活动期与静止期患者之间T细胞亚群上CD226^ 表达则无显著性差异（P＞0．05）。SLE患者CD3^ 、CD4^ 、CD8^ T细胞CD226^ 抗原表达水平与SLEDAI之间无明显相关性（P＞0．05）。结论：SLE患者体内存在T细胞亚群异常活化；活动期、静止期SLET淋巴细胞CD226^＋表达均增高，CD226^ 可能参与了SLE的免疫发病。     

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…     

重症肌无力患者外周血B7：CD28／CTLA4共刺激分子表达的动态变化

目的：研究重症肌无力（MG）患者B7：CD28／CTLA4共刺激通路相关分子表达动态变化及其与MG发病的关系。方法：用流式细胞仪检测18例MG患者和16例健康对照者外周血单个核细胞（PBMC）在经PMA（佛波酯）＋ionomycin(钙离子导入剂）刺激的0h,6h,24h,48h时B7－1、B7－2、CD28、CTLA4分子在CD4^ T细胞和CD8^ T细胞表面的表达。结果：（1）0h,MG患者B7－1、B7－2分子总体表达增加，CD4^ 、CD8^ T细胞表面B7－1、B7－2的表达未见明显增加，PMA＋ionomycin刺激后B7－1、B7－2在CD4^ 、CD8^ T细胞表面也无增加；（2）0h,CD28、CTLA4的表达增强，其中CD28^ 细胞的增多主要表现在CD4^ T细胞亚群，CTLA4的表达增强主要在CD8^ T细胞，在PMA＋ionomycin激活后，CD28表达明显增强，持续到48h都处于较高水平（与对照组相比P＜0．01），CTLA4表达出现短暂增强，6h即达高峰，以后下降，与对照组相比无显著增强（P＞0．05）。结论：MG患者外周血中B7：CD28／CTLA4通路共刺激相关分子表达增高，持续时间延长，检测共刺激分子的表达可反映机体的免疫激活状态，B7：CD28／CTLA4通路在MG发病中可能起重要作用。     

SLE患者外周血T细胞亚群ICOS与CD28共表达水平与疾病的关系

目的观察系统性红斑狼疮(systemic lupus erythematosus，SLE)病人外周血CD4^ 及CD8^ T细胞表面可诱导共刺激分子(inducible co-stimulator，ICOS)及CD28共表达水平，探讨ICOS和CD28共表达水平与SLE疾病的关系。方法采用三色流式细胞术检测SLE病人(n=51)及正常人(n=30)外周血CD4^ 及CD8^ T细胞表面ICOS与CD28共表达水平，并结合SLE病人疾病活动程度、临床表现等进行分析。结果与正常人相比，SLE活动期和稳定期病人外周血CD4^ T细胞中仅表达ICOS不表达CD28(即CD28^-ICOS^ )的细胞比例明显升高，但活动期病人和稳定期病人之间并无差异；活动期病人外周血CD8^ T细胞上同时表达CD28和ICOS的细胞(即CD28^ ICOS^ 细胞)和CD28^-ICOS^ 细胞比例都明显升高；同一SLE病人在疾病活动期CD4^ 和CD8^ T细胞中CD28^ ICOS^ 的细胞比例和CD8^ T细胞中CD28-ICOS^ 细胞比例明显高于该病人经过治疗病情稳定时；初发未治疗SLE病人仅CD4^ T细胞中CD28^ ICOS^ 细胞的比例比复发病人明显升高；血清抗dsDNA抗体( )病人和血清免疫球蛋白含量(IsG、IgA和IgM中任何一种)异常升高的病人外周血CD4^ T细胞中CD28^ ICOS^ 细胞和CD8^ T细胞中CD28-ICOS^ 细胞比例分别明显高于血清抗dsDNA抗体(-)和血清免疫球蛋白含量正常的病人。结论CD28和ICOS在SLE病人外周血CD4^ 和CD8^ T细胞上共表达水平与SLE疾病的活动程度、病程及临床表现等存在一定的关联。     

SLE患者Th1/Th2平衡状态及与外周血淋巴细胞CD28及CTLA-4分子表达的关系

目的:了解SLE患者Th1/Th2平衡状态以及共刺激分子CD28/CTLA-4与Th1/Th2平衡状态的关系。方法:研究对象为18例SLE患者(活跃期12例、缓解期6例)。对照组14例,为健康体检者。外周血单个核细胞(PBMCs)经梯度密度离心法分离后置于含PMA(5μg/L)及ionomycin(500μg/L)培养液中培养72 h。采用ELISA方法检测培养的PBMCs上清液中IFN-γ及IL-10的含量。应用流式细胞技术检测培养的淋巴细胞CD28及CTLA-4分子的表达。结果:活跃期SLE患者培养的PBMCs分泌IL-10的量(351.29 ng/L±153.31 ng/L)较对照组(254.48 ng/L±120.69 ng/L)有一定程度的升高,但差异无显著(P0.05),IFN-γ的分泌量(25.76 ng/L±16.09 ng/L)明显低于对照组(50.71 ng/L±27.92 ng/L,P0.05),IL-10/IFN-γ比值(18.74±13.77)明显高于对照组(6.66±4.95,P0.05)。培养前、后SLE患者CD3+及CD8+T细胞CD28分子表达量与对照组比较均无显著差异。培养前活跃期SLE患者CD3+T细胞CTLA-4分子表达量(0.79%+0.37%)较对照组(1.31%+0.61%)明显降低(P0.05)。培养后SLE患者CD3+T细胞及CD8+T细胞CTLA-4分子表达量仍低于对照组,但差异无显著(P0.05)。活跃期SLE患者培养的PBMCs中CD3+T细胞CTLA-4分子的表达量与上清液中IFN-γ含量呈明显的直线正相关关系(r=0.681,P0.05)、与上清液中IL-10及IL-10/IFN-γ比值呈明显的直线负相关关系(r=-0.624,P0.05;r=-0.738,P0.01)。结论:SLE患者存在Th1/Th2平衡向Th2方向偏移,即Th2优势状态。CTLA-4分子可能通过抑制CD28的信号转导参与Th2优势状态的形成。     

Aberrant expression of the costimulatory molecule CD40 ligand on monocytes from patients with systemic lupus erythematosus

CD40 ligand (CD40L, CD154) is overexpressed on T and B cells in systemic lupus erythematosus (SLE). Monocytes have been shown to contribute to immune-mediated pathology in SLE and to express CD40L under certain conditions. Therefore, we studied CD40L expression on lupus monocytes ex vivo, as well as after activation in vitro. A highly significant sevenfold increase in the frequency of CD40L-expressing peripheral monocytes from 23 SLE patients, compared to 16 healthy individuals (mean percentage of CD40L(+)CD14(+) among CD14(+) cells, 11.7 versus 1.6), was found by flow cytometry. Increased CD40L expression on monocytes correlated significantly with disease activity, elevated gamma-globulin serum levels, as well as increased CD40L expression on T cells. CD40L expression by lupus monocytes was verified at both the mRNA and protein levels, while LPS stimulation was found to upregulate CD40L mRNA accumulation and surface protein expression. CD40L expression on activated lupus monocytes within anti-CD3-stimulated, mononuclear cell cultures was also enhanced compared to control-derived monocytes. These novel findings underscore the multiplicity of pathways through which monocytes may contribute to SLE pathology and suggest that T cell-independent CD40L-mediated cell to cell interactions may be also involved in humoral immune activation in SLE.     

CD28 costimulation and CD28 expression in T lymphocyte subsets in HIV-1 infection with and without progression to AIDS

In a prospective study of 152 HIV-1 patients (with and without progression to AIDS) we examined CD28 MoAb costimulation and CD3 MoAb response using whole blood culture at baseline and up to either the time of AIDS diagnosis or the end of the observation period. CD28 antigen expression on both CD4+ and CD8+ T lymphocytes was also studied in both groups of patients. In patients who progressed to AIDS, CD28 MoAb costimulation was found to be decreased. Univariate time-dependent analysis showed that decreases in (i) absolute numbers of either CD4+, CD4+CD28+, CD8+CD28+ T cells, (ii) CD28 MoAb costimulation, and (iii) CD3 MoAb response, and an increase in CD8+CD28- %, are significant predictors for progression to AIDS. In addition, multivariate time-dependent analysis demonstrated that a decrease in CD28 MoAb costimulation (but not a decrease in CD3 MoAb response) was predictive for progression to AIDS, as were decreases in the percentage of CD4+ T cells and the absolute number of CD4+CD28+ T cells. Thus, CD28 MoAb costimulation can be considered a useful assay for monitoring HIV-1 infection. Furthermore, apart from the early increase in the percentage of CD8+CD28- T cells and an increase in the percentage of CD28- on CD8+ T cells in both groups of patients at baseline compared with normal controls, a negative correlation was found to exist between the percentages of CD4+ or CD4+CD28+ T cells and the percentage of CD8+CD28- T cells; this suggests that these cells are probably mutually regulated.     

IL-10 production in B cells is confined to CD154+ cells in patients with systemic lupus erythematosus

The immunological hallmark of SLE is B cell hyperactivity. CD154 (CD40-L) is normally expressed in activated T cells, and plays an important role in T-B interactions. Its expression is increased in SLE T cells. Additionally, its expression on B cells leads to the development of SLE-like disease in a transgenic model. IL-10 is a key cytokine in the disturbed SLE immune system. The aim of this work was to explore the relation between IL-10 and CD154 expression in SLE B cells. We studied 11 SLE patients and 10 healthy volunteers. Mononuclear cells were isolated from peripheral blood and cultured in the presence or absence of Cowan I Strain Staphylococcus (CSS). Surface CD154 and intracytoplasmic IL-10 expression were quantified with flow cytometry. In basal conditions, CD154 expression was not different in patients and controls. B cell stimulation did not cause a significant increase in CD154 expression in control B cells. However, its expression increased 2 times in B cells obtained from SLE patients. IL-10 expression was confined to CD154(+) cells. Our results show that IL-10 production is intimately linked to CD154 expression in B cells, and that the IL-10(+)CD154(+) B cell subset increases abnormally when SLE-derived cells are stimulated with CSS.     

Preferential elimination of CD28+ T cells in systemic lupus erythematosus (SLE) and the relation with activation-induced apoptosis

CD28 on T cells provides a potent costimulatory signal for T cell activation. Down-regulation of CD28 on peripheral T cells has been reported in certain clinical conditions, but full studies on the mechanism and biological significance have not been performed. Our extensive phenotype analysis of peripheral blood lymphocytes (PBL) from SLE patients revealed that the absolute number of CD28⁺ T cells of both CD4 and CD8 phenotypes was selectively decreased, while that of CD28⁻ T cells was maintained. CD28⁺ T cells from SLE patients exhibited mostly normal proliferative responses to both CD28-dependent and -independent stimulations. In contrast, CD28⁻ T cells were hyporesponsive to anti-CD3 stimulation in both SLE and normal controls. These results implied that the selective decrease of CD28⁺ T cells in SLE does not result from a hyporesponsiveness of CD28⁺ T cells. To investigate the reason for the selective loss of CD28⁺ T cells, we determined the appearance of apoptotic cells in culture with or without anti-CD3 stimulation. Apoptotic cells defined by merocyanine (MC)540 were gradually increased from 12 h to 24 h. Anti-CD3-induced apoptosis of CD28⁺ T cells was significantly accelerated in SLE, whereas apoptosis of CD28⁻ T cells was hardly detected in both SLE and normal controls. Comparative analysis between CD28⁺ and CD28⁻ T cells on CD95 (Fas) and Bcl-2 expression, which are related to activation-induced cell death (AICD), did not show a major difference, although CTLA4, which has been demonstrated to transmit an apoptosis-inducing signal, was expressed only on CD28⁺ T cells. Our results suggest that CD28-mediated costimulation influences T cell susceptibility to AICD and may be involved in T cell lymphopenia in SLE.     

Glucocorticosteroid therapy decreases CD14-expression and CD14-mediated LPS-binding and activation of monocytes in patients suffering from systemic lupus erythematosus

In order to study the possible action of glucocorticosteroids (GCS) on the CD14/Toll like receptor mediated activation of monocytes the CD14-expression, CD14-mediated LPS binding and activation of these cells of patients suffering from Systemic Lupus Erythematosus receiving no, low dose or pulse steroid treatment was studied. The CD14-expression was determined on whole blood monocytes by flow cytometry, while the LPS-binding of an FITC-LPS preparate and the LPS-induced TNFalpha secretion were tested on isolated monocytes. The CD14-dependent and -independent LPS-binding and activation were evaluated with the help of a blocking anti-CD14 mAb. Our results showed that the CD14-expression, CD14-dependent LPS-binding and activation were significantly inhibited by the in vivo applied pulse steroid therapy. In contrast, the CD14-independent LPS-binding and activation were not altered by the GCS treatment. Our data provide further in vivo evidence for a possible new way of GCS therapy is able to initiate its anti-inflammatory action.     

Human cytomegalovirus in patients with systemic lupus erythematosus

The objective of this study was to determine the frequencies of human cytomegalovirus (HCMV) infection and HCMV genome copy number in blood of consecutive (treated from several months to several years) systemic lupus erythematosus (SLE) patients (22 women). The obtained results were compared to the healthy controls (15 women). All patients fulfilled at least four of the 1982 revised American rheumatism association (ARA) classification criteria for SLE. Our patients demonstrated three or four of the nine possible organ systems involved and most of them had mild SLE with SLE disease activity index (SLEDAI) score < 10 at time when blood samples were collected to detect HCMV. Quantitative analysis of HCMV genome was performed with aid of sequence analyzer ABI PRISM ^TM 7700 Perkin Elmer. Primers and probe were constructed on the basis of IE4 region of HCMV genome. The viral load was expressed as log₁₀ of calculated HCMV genome copy number. Qualitative analysis revealed that 100% of our SLE patients were infected with HCMV, whereas in the control group only 73% of persons were HCMV positive. Statistically significant difference was demonstrated when the strength of the association between SLE or controls and infection of HCMV was calculated (estimated by Fisher's exact test, P value = 0.02). Higher viral DNA copy number was observed in whole blood of SLE patients than in the control group (338.45 ± 221.76 and 229.00 ±405.61 copies/ml respectively) but did not reach statistical significance level (95% confidence interval from 170.41 to 249.32, P = 0.71). Furthermore percentage of patients with HCMV-DNA copy number >2.0 × 10² copies/ml was statistically significantly higher than this one in controls. The data show association between HCMV infection and SLE, which should be taken into account during the course of SLE.