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。     

外周血CD19+B细胞PD-L1的表达与系统性红斑狼疮发病的相关性分析

目的:探讨程序性死亡配体1(Programmed death ligand-1,PD-L1)在系统性红斑狼疮(Systemic lupus erythema-tosus,SLE)患者外周血B细胞上的表达及临床意义。方法:应用流式细胞仪检测51例SLE患者和38例健康对照者外周血CD19+B细胞表面PD-L1的表达水平,比较SLE稳定组、活动组和健康对照组以及狼疮肾炎组和无狼疮肾炎组之间CD19+B细胞表面PD-L1表达阳性细胞的百分比,并分析其与临床表现及实验室检查数据的相关性。结果:SLE活动组和稳定组CD19+PD-L1+B细胞百分率均低于健康对照组,活动组又低于稳定组,差异均有统计学意义(均P<0.05)。狼疮肾炎患者CD19+PD-L1+B细胞百分率低于无狼疮肾炎患者(P<0.05)。SLE患者CD19+PD-L1+B细胞百分率与SLEDAI评分、尿蛋白定量、呈负相关,与C3呈正相关。SLE患者中抗dsDNA抗体、抗Sm抗体、抗U1snRNP抗体、抗核小体抗体阳性组外周血B细胞PD-L1表达水平均低于对应阴性组,且均有统计学意义(均P<0.05)。结论:SLE患者外周血CD19+B细胞表达PD-L1下降,与病情活动性和抗体产生有很好的相关性。     

CD40在系统性红斑狼疮外周血淋巴细胞的表达

使用密度离心法分离SLE患者和正常人外周血单个核细胞 (PBMC ) ,采用流式细胞术检测B淋巴细胞白细胞分化抗原 4 0 (CD4 0 )的表达水平 ,进行SLE患者 (活动期和缓解期 )和正常人之间的比较 ;并进行B淋巴细胞CD4 0表达水平和血清抗dsDNA抗体水平及狼疮活动指数 (SLEDAI)的相关分析。结果表明 ,活动期SLE患者外周血B淋巴细胞比例 (% )和其表达CD4 0的比例 (% )均明显高于缓解期SLE患者和对照组 ,其表达CD4 0的平均荧光强度 (MFI)在活动期SLE患者最高 ,缓解期SLE患者稍低 ,对照组最低 ;相关分析结果表明 ,活动期SLE患者B淋巴细胞CD4 0的表达比例 (% )和强度 (MFI)均与血清抗dsDNA抗体及SLEDAI呈正相关 ,后两者呈高度相关 ;缓解期SLE患者B淋巴细胞表达CD4 0的强度 (MFI)和SLEDAI呈正相关。CD4 0在活动期SLE患者B淋巴细胞的表达增加 ,其水平与疾病活动度有关。     

系统性红斑狼疮患者外周血CD14~+单核细胞表达PD-L1的分析和意义

目的:探讨PD-L1在系统性红斑狼疮(SLE)患者外周血单核细胞(Mo)上的表达及临床意义。方法:应用流式细胞仪检测51例SLE患者和38例健康对照者外周血CD14+Mo表面PD-L1表达水平,比较SLE不活动组、活动组和健康对照组以及狼疮肾炎组和无狼疮肾炎组之间CD14+Mo表面PD-L1表达的百分比,并分析其与临床表现及实验室检查数据的相关性。结果:SLE活动组和稳定组CD14+PD-L1+Mo百分率均高于健康对照组,差异均有统计学意义(P<0.05)。狼疮肾炎患者CD14+PD-L1+Mo百分率高于无狼疮肾炎患者(P<0.05)。SLE患者CD14+PD-L1+Mo百分率与SLEDAI评分、尿蛋白定量呈正相关。SLE患者中抗dsDNA抗体、抗Sm抗体、抗U1snRNP抗体、抗核小体抗体阳性组外周血CD14+PD-L1+单核细胞百分率均高于对应阴性组,且均有统计学意义(均P<0.05)。结论:SLE患者外周血CD14+Mo细胞表达PD-L1异常,与病情活动性和抗体产生有关。     

系统性红斑狼疮患者B淋巴细胞EBV-LMP1和ZEBRA的表达研究

目的：探讨ＥＢＶ－ＬＭＰ１和ＺＥＢＲＡ在系统性红斑狼疮患者（ＳＬＥ）的表达情况。方法：间接荧光免疫标记，流式细胞仪检测。结果：ＳＬＥ患者Ｂ淋巴细胞中ＥＢＶ－ＬＭＰ１和ＺＥＢＲＡ的表达显著高于正常对照组（Ｐ＜０．０１）。活动期患者ＣＤ２３＋细胞ＥＢＶ－ＬＭＰ１和ＺＥＢＲＡ的表达率均高于ＣＤ１９＋细胞（Ｐ＜０．０１）。非活动期患者ＣＤ２３＋细胞ＥＢＶ－ＬＭＰ１表达也高于ＣＤ１９＋细胞（Ｐ＜０．０１）。但ＥＢＶ－ＺＥＢＲＡ表达在两亚群间差异没有统计学意义（Ｐ＞０．０５）。结论：朋病毒参与了ＳＬＥ的发病机制，病毒主要以潜伏期状态存在于患者中，病毒复制促进病情发展，检测Ｂ淋巴细胞ＥＢＶ－ＬＭＰ１和ＺＥＢＲＡ的表达率，有助于病情活动指标的判断。     

系统性红斑狼疮患者外周血单个核细胞TRAF1 mRNA的表达及其意义

目的：研究肿瘤坏死因子受体相关因子1(TRAF1)mRNA表达在SLE患者的外周血单个核细胞中的表达是否异常，以及该基因mRNA表达与SLE活动指数是否相关。方法：TRAF1 mRNA表达采用RT-PCR半定量法；SLE活动指数以SLEDAI为判断标准。结果：TRAF1 mRNA在SLE患者的外周血单个核细胞中的表达较正常人为高，但与SLEDAI指数无相关。结论：TRAF1基因可能在SLE的发病机制中起重要作用。     

儿童系统性红斑狼疮患者外周血淋巴细胞表面CD95的表达及临床意义

目的:探讨儿童系统性红斑狼疮(systemic lupus erythematosus,SLE)外周血淋巴细胞表达CD95的特征及与疾病活动性和其他免疫学指标间的关系.方法:使用流式细胞术检测60例SLE患儿和20例对照外周血T淋巴细胞亚群和B淋巴细胞表面CD95的表达,并分析其与SLE疾病活动性以及实验室检查之间的关系.结果:初发SLE患儿外周血中CD4+T细胞表面CD95的表达显著高于对照组,差异有统计学意义(P<0.05);初发SLE患儿外周血中CD19+B细胞表面CD95的表达显著高于健康儿童(P<0.05);CD19+CD95+B细胞的比例和SLE疾病活动性呈正相关(r=0.4,P<0.05);CD4+CD95+T细胞的比例和SLE疾病活动性呈正相关(r=0.3,P<0.05),CD4+CD95+T细胞的比例和外周血抗双链DNA抗体(anti-dsDNA Abs)的水平呈正相关(r=0.2,P<0.05);治疗后SLE患儿外周血中CD19+CD95+B细胞和CD4+CD95+T细胞的比例均有显著下降,差异有统计学意义(P<0.05).结论:儿童SLE患者外周血中淋巴细胞表达CD95的水平显著升高,且与SLE的疾病活动性及血清中抗双链DNA抗体相关,可以作为SLE的评价指标.     

DNAM-1在系统性红斑狼疮患者T淋巴细胞的表达研究

目的：研究DNAM－1在SLE患者外周血T淋巴细胞亚群上的表达，以阐明DNAM－1抗原在SLE患者体内活化作用以及与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PHA刺激培养72小时后，三色荧光标记的单克隆抗体染色，利用流式细胞仪测定T淋巴细胞亚群膜表面DNAM－1抗原的表达。同时检测SLE患者抗dsDNA抗体、C3和C4补体，疾病活动性用SLEDAI记分。结果：SLE患者CD4^＋、CD8^＋淋巴细胞上DNAM－1表达率均高于正常对照组（P＜0．01）；活动期SLE组CD4^＋、CD8^＋细胞上DNAM－1表达高于正常对照组和静止期SLE组（P＜0．01），而静止期SLE组与正常对照组无明显性差异（P＞0．05）；SLE患者CD8^＋细胞DNAM－1表达与SLEDAI、抗dsDNA抗体之间呈显著正相关（P＜0．001），与C3和C4补体水平呈明显负相关（P＜0．05），CD4^＋细胞DNAM－1表达与SLEDAI、抗dsDNA抗体、C3和C4补体之间无明显相关性（P＞0．05）。结论：SLE患者内存在T细胞亚群异常活化；活动期SLE患者T淋巴细胞亚群DNA－1表达增高；SLE患者CD8^＋细胞DNAM－1表达异常与SLEDAI、抗dsDNA抗体、C3和C4补体之间有明显相关，CD8^＋细胞活化程度可能与SLE疾病严重程度有关，故DNAM－1可能参与了SLE的免疫发病机理。     

系统性红斑狼疮病人T和B淋巴细胞凋亡的观察

为了研究SLE的T、B淋巴细胞及其亚类的凋亡情况,采用碘化丙锭染色,在流式细胞仪下观察计数:22例SLE病人淋巴细胞(包括总体淋巴细胞、CD3~+、CD4~+、CD8~+T细胞和CDl9~+B细胞)凋亡率在培养0、24、48h时均较正常组有显著增高,并以CD4~+T细胞和CDl9~+B细胞在活动期SLE病人中凋亡更突出。此外,SLE病人,自身抗体产生愈多,其细胞凋亡率愈高;疾病活动度增高,凋亡率也较高。提示;SLE病人的淋巴细胞凋亡在体外加速,与其自身抗体产生有密切关系,在其发病机制中起一定的作用。     

冠心病患者外周血T淋巴细胞PD-L1的表达及意义

目的研究程序性死亡配体-1（PD-L1）在冠心病患者外周血中T淋巴细胞上的表达及意义。方法按照WHO关于冠心病诊断标准,将实验分为稳定型心绞痛（SA）组（n=23）、急性冠脉综合征（ACS）组（n=21）和正常对照组（n=18）,三组均经冠状动脉造影证实。采集三组人群的外周血,用流式细胞技术及RT-PCR技术测定T淋巴细胞PD-L1蛋白及mRNA表达情况。结果与正常对照组比较,SA组和ACS组外周血T淋巴细胞PD-L1蛋白及mRNA表达水平均显著升高（P〈0.01）;但SA组与ACS组比较差异无统计学意义（P〉0.05）。结论冠心病患者外周血T淋巴细胞PD-L1表达上调,其可能在动脉粥样硬化斑块形成过程中起作用。     

Expression of CTLA-4 Molecule in Peripheral Blood T Lymphocytes from Patients with Systemic Lupus Erythematosus

CTLA-4 is a cell surface molecule expressed on activated T cells that is suggested to deliver a negative signal for T cell activation. Since CTLA-4 might be a negative regulator of autoimmune diseases, we investigated its expression on T cells from 20 patients with systemic lupus erythematosus (SLE) by flow cytometric analysis and RT-PCR. We found that although CTLA-4 mRNA was readily detected in all patients and controls, only a very minor subset of T cells expressed detectable surface CTLA-4 molecules in both groups. But patients with SLE had significantly increased percentages of CTLA-4-positive T cells compared with normal controls, implying at least that there was no apparent defective expression of CTLA-4 molecule in human lupus. The kinetics of CTLA-4 expression on T cells stimulated in vitro with PMA plus ionomycin were similar in normal controls and patients with SLE. The expression of CTLA-4 molecules after stimulation increased gradually and peaked at 72 hr. However, the induction of CTLA-4 expression on patients' T cells appeared to be weaker than that of normal individuals. Whether this reflects impaired down regulation by CTLA-4 molecules in SLE patients needs to be clarified further.     

Expression of CD80 and CD86 on Peripheral Blood T Lymphocytes in Patients with Systemic Lupus Erythematosus

CD80 and CD86 were detected in high amounts on circulating T cells in the peripheral blood of some patients with systemic lupus erythematosus (SLE), using flow cytometry and monoclonal antibodies. Patients with other connective tissue diseases did not have a high percentage of T cells expressing CD80 or CD86 in their peripheral blood. CD80 was expressed mainly on CD4 T cells, whereas CD86 was expressed on CD8 T cells, and these two populations were associated with paticular clinical features. These two molecules were expressed on different T-cell populations and might have different roles in the generation and regulation of immune responses. Since high expression of CD86 on T cells was detected much earlier than the appearance of clinical features and a high titer of anti-DNA antibody, it may be a useful parameter for predicting the flare of SLE.     

In Vitro TNP-Specific Antibody Formation by Peripheral Lymphocytes from Patients with Systemic Lupus Erythematosus

Immunological reactivity in patients with systemic lupus erythermatosus (SLE) was assessed by investigating in vitro trinitrophenyl (TNP)-specific antibody formation by peripheral lymphocytes. Peripheral lymphocytes from 16 patients with SLE were cultured with TNP conjugated with horse erythrocytes (TNP-HRBC) in the presence of 2-mercaptoethanol. The hemolytic plaque assay was used to detect hapten (TNP)-specific antibody-forming cells. Peripheral lymphocytes from normal individuals failed to produce antibody to TNP, whereas SLE lymphocytes produced a significant number of plaque-forming cells. Co-culture experiments with SLE and normal lymphocytes suggested that patients with SLE have a defect in T lymphocytes, leading to abnormal antibody production.     

Mitogenic Responses to Lipopolysaccharide by B Lymphocytes from Patients with Systemic Lupus Erythematosus

Peripheral blood lymphocytes from 43 patients with systemic lupus erythcmatosus (SLE) and from age- and sex-matched normal controls were cultured with lipopolysaccharide (LPS) to examine the response to the polyclonal B-cell activator. Lymphocytes from active SLE patients incorporated 4840±471 (mean ± SE) cpm in response to LPS, whereas lymphocytes from inactive SLE patients incorporated 6906 ± 897 cpm. In contrast, lymphocytes from normal individuals incorporated 7452 ± 1126 cpm. Ig synthesis of lymphocytes from active SLE in response to LPS stimulation was also less than that of normal individuals. The helper T-cell function of active SLE, as examined by co-culturing irradiated SLE lymphocytes with unirradiated normal lymphocytes, was normal. These results thus suggested that a defect of B lymphocytes exists in active SLE patients. This B-cell defect and T suppressor cells apparently play an important role in the pathogenous of SLE.     

Activation Profile of Intracellular Mitogen-Activated Protein Kinases in Peripheral Lymphocytes of Patients with Systemic Lupus Erythematosus

Introduction  

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease associated with aberrant activation of T and B lymphocytes. Abnormal activation of intracellular signaling molecules in lymphocytes by inflammatory cytokines can instigate the inflammation in SLE.     

系统性红斑狼疮病人环孢素A受体mRNA的表达

目的探讨静止期、活动期系统性红斑狼疮病人(SLE)外周血单个核细胞(PBMC)环孢素A受体(CyP) mRNA表达及糖皮质激素对其表达的影响,为临床应用环孢素A辅助治疗该病提供理论依据.方法采用逆转录PCR方法,经凝胶图像半定量分析,检测患者外周血单个核细胞CyP mRNA的表达.结果 SLE病人外周血单个核细胞存在有CyP mRNA的表达,静止期较正常人差异无显著性(p>0.05)意义,但在活动期CyP mRNA的表达较正常人和静止期病人明显降低(p<0.01),地塞米松处理后,活动期病人CyP mRNA的表达水平显著上升(p<0.01).结论 SLE病人有CyP mRNA的表达,但活动期明显下降,糖皮质激素可改善CyP mRNA的表达.     

Differential Expression of CD30 on CD3 T Lymphocytes in Patients with Systemic Lupus Erythematosus

Systemic lupus erythematosus (SLE) is an autoimmune systemic disease caused as a result of an imbalance of Th1‐/Th2‐type cytokines. The soluble form of CD30 (CD30s) released from peripheral blood cells has been described as a marker of active disease in Th2‐type immune response as in SLE. However, the expression of CD30 on CD3 T lymphocytes from patients with SLE has not been studied yet. Therefore, we have addressed our study to attempt this issue, studying CD30 expression by flow cytometry on CD3 T lymphocytes and CD4/CD8 subsets in samples from SLE patients mainly with lupus nephritis. In parallel, we have determined the production of the cytokines IL‐4 (Th2), IFNγ (Th1), IL‐10 and TGFβ by intracellular staining. Differences between positive CD30 T cells in healthy controls and patients with SLE were found, with a higher percentage of CD30‐expressing T cells in patients with SLE (P = 0.001). In contrast to healthy controls, CD30 was mainly expressed on CD8 T cells from patients with SLE. The intracellular cytokine staining showed that TGFβ is the main cytokine expressed in CD3 T cells from patients with SLE. In addition to this, we have found a positive correlation between CD30‐expressing T cells and IL‐4, IFNγ, and immunosuppressive cytokines (IL‐10 and TGFβ) (P < 0.05). These results suggest that CD30 could play a role in the pathogenesis of SLE and its expression on CD3 T lymphocytes is not restricted only to Th2‐type response.