窒息新生大鼠血浆ET水平及L-Arg和L-NAME对其影响

为探讨一氧化氮（ＮＯ）的前体Ｌ－精氨酸（Ｌ－Ａｒｇ）和ＮＯ合成酶抑制剂Ｌ－硝基－精氨酸甲酯（Ｌ－ＮＡＭＥ）对围产期窒息新生大鼠血浆内皮素（ＥＴ）的影响,采用放射免疫分析法检测窒息新生大鼠及窒息前应有Ｌ－Ａｒｇ或Ｌ－ＮＡＭＥ预处理的新生鼠血浆ＥＴ的水平。结果显示：窒息组血浆ＥＴ水平较对照组明显升高（Ｐ〈０．０１）;Ｌ－Ａｒｇ组血浆ＥＴ水平较窒息组明显下降（Ｐ〈０．０１）,与对照组相比无显著差异（Ｐ〉     

系统性红斑狼疮病人血T,B淋巴细胞Bcl—2的表达

目的：探讨Ｂｃｌ－２在系统性红斑狼疮（ＳＬＥ）发病机制中作用。方法：采用流式细胞仪双标记法检测３１例ＳＬＥ病人外周血Ｔ、Ｂ细胞Ｂｃｌ－２蛋白表达。结果：发现活动期ＳＬＥ病人活动期ＳＬＤ病人ＣＤ３＾＋、ＣＤ４＾＋和ＣＤ８＾＋Ｔ细胞Ｂｃｌ－２蛋白表达明显高于非活动期ＳＬＥ病人、其他疾病组和正常对照组。ＣＤ１９＾＋Ｂ细胞Ｂｃｌ－２蛋白表达在各组之间并无统计学差异。ＣＤ３＾＋Ｔ细胞Ｂｃｌ－２蛋白表达的平均     

SLE患者血液SC5b—9,ANA及AdsDNA浓度的测定及意义

目的：对５３例ＳＬＥ患者血液中的补体末端复合体（Ｃ５ｂ－９）、补体溶血总活性（ＣＨ５０）、抗核抗体（ＡＮＡ）及抗双链ＤＮＡ（ＡｄｓＤＮＡ）抗体的浓度进行了测定。结果：３３例活动期ＳＬＥ患者，其ＳＣ５ｂ－９、ＡＮＡ及ＡｄｓＤＮＡ的水平均较２０例非活动期ＳＬＥ患者高，ＣＨ５０活性则较非活动期ＳＬＥ患者低，活动期与非活动期ＳＬＥ患者４项指标间的差异均显著（Ｐ＜００１）。此外，用不同方法检测各项指标的敏感性以ＳＣ５ｂ－９敏感性最高。结论：患者ＳＣ５ｂ－９、ＡｄｓＤＮＡ、ＡＮＡ和ＣＨ５０指标的变化，可为临床正确判断ＳＬＥ患者的病情活动，指导用药及评估预后等提供科学依据     

红细胞抗原免疫印迹法在抗SSA抗体分型中的应用

建立了以人红细胞为ＳＳＡ抗原来源的免疫印迹法（ＲＢＣ－ＩＢＴ）检测ＳＳＡ抗体。此法的敏感性略低于常规的对流免疫电泳法（ＣＩＥ），但可检出部分ＣＩＥ中阴性的ＳＳＡ抗体阳性血清，而且尚可鉴定出ＳＳＡ抗体的两种亚型（５２ＫＤ和６０ＫＤ）。检测了干燥综合症（ＳＳ）、系统性红斑狼疮（ＳＬＥ）及类风湿性关节炎（ＲＡ）等三组病人血清，其中ＳＳ和ＳＬＥ两组在ＳＳＡ抗体亚型的分布上差异显著。抗５２ＫＤＳＳＡ抗体主要分布于ＳＳ组（Ｐ＜０．０５），而抗６０ＫＤＳＳＡ抗体主要见于ＳＬＥ组（Ｐ＜０．０５），ＲＢＣ－ＩＢＴ中，非特异或不明显色带极少见，而ＳＳＡ抗体带清晰、易辨。     

SLE患者CD45RA^＋和CD45RO^＋T细胞亚群的变化

利用流式细胞术及免疫双荧光染色法，检测３５例系统性红斑狼疮（ＳＬＥ）患者外周血ＣＤ４５ＲＡ和ＣＤ４５ＲＯＴ细胞亚群，结果表明：ＳＬＥ患者ＣＤ４细胞降低，ＣＤ８细胞增高，ＣＤ４／ＣＤ８降低，ＣＤ４ＣＤ４５ＲＡ细胞在病情活动期降低，在稳定期回升，ＣＤ４ＣＤ４５ＲＯ细胞在活动期有增高倾向，在稳定期下降，ＣＤ８ＣＤ４５ＲＡ及ＣＤ８ＣＤ４５ＲＯ细胞均在活动期增高，又均在稳定期降至正常水平，提示ＣＤ４５ＲＡ和     

间变性大细胞性T细胞性淋巴瘤中EB病毒检测及CD56的表达

目的：研究间变性大细胞性Ｔ细胞性淋巴瘤ＥＢ病毒表达情况及其与ＣＤ５６阳性表达间的关系。方法：采用免疫组织化学ＬＳＡＢ法检测１５例ＡＬＴＣＬ中Ｋｉ－１及ＣＤ５６的表达,并用原位杂交法检测其ＥＢＥＲｓ。结果：１５例ＡＬＴＣＬ中Ｋｉ－１均阳性（１００％）,５例ＣＤ５６阳性（３３．３％）,９例ＥＢＥＲＳ阳性。其中,３例ＡＬＴＣＬ中ＥＢＥＲＳ和ＣＤ５６共同阳性。结论：ＡＬＴＣＬ的发生同ＥＢ病毒感染有一定关系     

拮抗内皮素生物学效应对甘油所致大鼠急性肾功能衰竭的影响

目的和方法：在甘油致大鼠急性肾功能衰竭（ＡＲＦ）模型上,观察外源内皮素（ＥＴ）以及心房钠尿肽（ＡＮＰ）、硝苯吡碇（Ｎｉｆ）和ＮＯ前体Ｌ－精氨酸（Ｌ－Ａｒｇ）的作用。结果：甘油致大鼠ＡＲＦ后２４ｈ,肾功能明显受损,血清尿素氮（ＳＵＮ）,肌酐（Ｓｃｒ）,丙二醛（ＭＤＡ）,肾皮质钙含量及血浆内皮素（ＰＥＴ）水平均明显高于正常对照组（Ｐ＜０００１）。外源性ＥＴ可加重ＡＲＦ大鼠肾损伤,使ＳＵＮ、Ｓｃｒ、ＭＤＡ、ＰＥＴ及肾皮质钙含量显著增加,而ＡＮＰ、Ｎｉｆ和Ｌ－Ａｒｇ则使ＡＲＦ大鼠ＳＵＮ、Ｓｃｒ、ＭＤＡ、ＰＥＴ及肾皮质钙含量明显降低,肾组织ｃＧＭＰ含量增加,减轻肾小管损伤。结论：ＥＴ对ＡＲＦ的发生发展具有重要意义,ＡＮＰ、Ｎｉｆ和Ｌ－Ａｒｇ可通过不同途径拮抗ＥＴ的生物学效应,从而对ＡＲＦ的防治起到积极的作用。     

银杏叶制剂对蛛网膜下腔出血大鼠血浆及脑组织内皮素—1含量？…

目的 探讨蛛网膜下腔出血（ＳＡＨ）后内皮素－１（ＥＴ－１）变化和银杏叶制剂（ＧＢＥ）对其影响。方法 对单纯ＳＡＨ组和ＧＢＥ处理组大鼠检测ＳＡＨ后２４ｈ内颅内血浆及脑组织ＥＴ－１含量的动脉改变。结果 ＳＡＨ组血浆及脑组织ＥＴ－１含量分别于ＳＡＨ后即刻和１ｈ明显增加,并维持２４ｈ（Ｐ〈０．０５,０．０１）。ＧＢＦ组ＳＡＨ后ＥＴ－１增加的程度显著小于ＳＡＨ组（Ｐ〈０．０５,０．０１）。结论 血浆及脑组织     

磷脂抗体阳性红斑狼疮患者周围血T细胞受体基因表达初探

目的：初步探讨ＳＬＥ患者周围血Ｔ细胞受体Ｖβ基因是否存在偏移。方法：应用荧光ＰＣＲ定量方法对ＨＬＡ－ＤＲＢ１＾＊０８０３／ＤＱＡ１＊０１０３的抗磷脂抗体阳性ＳＬＥ患者４例,病人对照干燥综合征（ＳＳ）１例,正常对照３例进行周围血ＴＣＲ－Ｖβ的分析。结果;健康者和病人对照的ＴＣＲ仅少数Ｖβ有轻微增高表达。１例非活动性ＳＬＥ患者与对照组相仿,但３例活动性ＳＬＥ５的ＴＣＲ－Ｖβ基因谱呈多克隆激活,其中２例     

开博通与左旋精氨酸对自发性高血压大鼠血管内皮细胞功能影响

目的通过一氧化氮（ＮＯ）和内皮素（ＥＴ）的改变观察开博通和左旋精氨酸对自发性高血压大鼠（ＳＨＲ）血管内皮细胞功能影响,以探索两者联合使用的降压机制。方法Ｗａｓｔａｒ大鼠 １０Ｒ作正常对照组。３８只 ＳＨＲ随机分为 ４组： ＳＨＲ对照组（ｎ＝１０）,Ｌ－Ａｒｇ治疗组（ｎ＝１０）,开博通治疗组（ｎ＝９）,Ｌ－Ａｒｇ＋开博通联合治疗组（ｎ＝９）。同时测各组用药前后的血压,一氧化氮（ＮＯ）和内皮素（ＥＴ）。结果ＳＨＲ组的ＮＯ低于正常对照而ＥＴ则高于对照组（Ｐ＜ ０．０５）,Ｌ－Ａｒｇ和／或ＣＰＴ两治疗组与ＮＯ和ＥＴ各有不同程度升高和降低．（与ＳＨＲ组比较均Ｐ＜０．０５）,其中以Ｌ－Ａｒｇ＋ＣＰＴ联合治疗组最明显。３个治疗组用药后ＢＰ较用药前均有不同程度的下降（均Ｐ＜０．００１）,而又以Ｌ－Ａｒｇ ＋ＣＰＴ联合组降压效果最佳。结论ＣＰｔ和Ｌ－Ａｒｇ均可有效增加 ＳＨＲ的 ＮＯ合成释放,同时又加强 ＣＰＴ降压效应,由此降低血压进而改善血管内皮细胞功能。     

抗内皮细胞抗体和血小板生成素测定在ITP与SLE鉴别中的意义

目的：探讨抗内皮细胞抗体(AECA)和血小板生成素(TPO)测定在鉴别特发性血小板减少性紫癜（ITP）和系统性红斑狼疮（SLE）中的临床意义。 方法: 用ELISA法分别测定76例ITP患者、41例SLE患者及50例正常人血清中的AECA和TPO水平。 结果: SLE组、ITP组患者血清AECA水平明显高于正常对照组（P<0.01）；SLE组患者血清AECA水平显著高于ITP组（P<0.01）；ITP组患者血清TPO水平与正常对照组无显著差异(P>0.05)，而SLE组血清TPO水平显著高于ITP组患者和正常对照组(P<0.01)。 结论: 血清AECA和TPO的测定在鉴别诊断ITP和SLE中有显著的临床意义。     

Elevated serum anti-endothelial cell autoantibodies titer is associated with lupus nephritis in patients with systemic lupus erythematosus.

BACKGROUND AND PURPOSE: Systemic lupus erythematosus (SLE) is an autoimmune connective tissue disease associated with endothelial dysfunction and the existence of multiple species of autoantibodies. However, the association between endothelial dysfunction and renal manifestations remains unclear in Taiwanese SLE patients. METHODS: Serum samples were collected from SLE patients with biopsy-proven lupus nephritis (n = 32), stable SLE patients (n = 32) and healthy controls (n = 32). The SLE Disease Activity Index (SLEDAI) of SLE patients was scored, and levels of anti-endothelial cell antibodies (AECA) and anti-endothelial activities in serum samples were measured by cell-enzyme-linked immunosorbent assay and crystal violet assay, respectively, using cultured human endothelial EA.hy926 cells. RESULTS: Significantly higher AECA (p<0.001) and anti-endothelial activities (p<0.001) were found in sera from patients with lupus nephritis compared with that from stable SLE patients or controls. Moreover, AECA titers (p<0.001) and anti-endothelial activities (p<0.001) were strongly correlated with SLEDAI scores in these patients. CONCLUSION: The strong correlations of AECA and anti-endothelial activity with lupus nephritis activity support an endothelial origin for renal complications in Taiwanese SLE patients.     

Characterization of the Endothelial Surface Proteins Recognized by Anti-Endothelial Antibodies in Primary and Secondary Autoimmune Vasculitis

The antigenic structures recognized by anti-endothelial cell antibodies (AECA) in sera from 10 Wegener's granulomatosis (WG) and 12 systemic lupus erythematosus (SLE) patients with signs of vasculitis were characterized by immunoprecipitation of selectively radiolabeled surface membrane proteins from human umbilical vein endothelial cells. Electrophoretic analysis of the immunoprecipitated proteins revealed reactivities against endothelial antigens ranging in size from 200 to 25 kDa. AECA antigens were not cell specific, since the same sera also reacted, at least in part, with radiolabeled human fibroblast surface proteins. The majority of WG patients displayed a constant precipitation pattern of five proteins (180, 155, 125, 68, and 25 kDa). On the contrary, AECA from SLE sera reacted with a more heterogeneous series of endothelial proteins. A group of four proteins, however, was also found in the majority of SLE sera: 200, 180, 155, and 25 kDa. In addition, some endothelial antigens were immunoprecipitated only by WG (125 kDa) or by SLE sera (200 kDa), suggesting a different endothelial reactivity in different vasculitic processes. The reaction did not involve intracellular proteins as demonstrated by the lack of reactivity of SLE sera negative for AECA but positive for anti-cytoplasmic or anti-nuclear antibodies. These data confirming that AECA recognize surface endothelial determinants further support a potential pathogenetic role for these antibodies in autoimmune vasculitis.     

Analysis of natural and disease-associated autoantibody repertoires: anti-endothelial cell IgG autoantibody activity in the serum of healthy individuals and patients with systemic lupus erythematosus

The present study demonstrates that natural IgG with anti-endothelialcell activity is present in the serum of healthy individualsand in pooled normal human Ig. By using a novel method thatallows for the simultaneous and quantitative assessment of reactivitiesof antibodies with a large number of antigens in tissues, weobserved that natural anti-endothelial cell antibody (AECA)recognizes a restricted set of self antigens in endothelialcells that is conserved among healthy individuals. The extentto which natural AECA activity is expressed in serum and thepattern of reactivity of AECA with endothelial cell antigensshowed little variability between individuals. Analysis of AECAin the serum of patients with systemic lupus erythematosus (SLE)revealed a higher amount of activity and a wider spectrum ofantigentic specificities than that recognized by natural antibodiesin endothelial cell extracts. AECA activity of IgG in wholeserum was lower than that of purified IgG in the case of healthyindividuals and showed little variation among individuals. incontrast, no difference was found between AECA activity of purifiedIgG and that of IgG in patients' serum, suggesting that SLEsera lack the factors that control expression of AECA activityin the serum of healthy individuals. Our results indicate thatnatural autoantibodies recognize a restricted and conservedset of self antigens. Our observations further suggest thatdefective regulation of the expressed autoreactive B cell repertoireis the basis for expansion of novel clonal specificities andenhanced autoantibody activity in serum of patients with autoimmunedisease.     

Antibodies to endothelial cells in systemic lupus erythematosus: a potential marker for nephritis and vasculitis

Using an ELISA, anti-endothelial cell antibodies (AECA) have been found in sera obtained at the time of renal biopsy in 46 out of 57 patients (81%) with systemic lupus erythematosus (SLE) and nephritis (mean binding index (BI) = 84% +/- 52.8) compared with 22 out of 50 SLE patients (44%) without nephritis (mean BI = 45% +/- 35.9). Seventy normal human sera had a mean BI of 10% +/- 9.8. The highest levels were seen in patients with diffuse proliferative glomerulonephritis (WHO grade IV) and in patients with proteinuria and nephrotic syndrome. When the biopsies were assessed for activity and chronicity scores, AECA were associated with active renal lesions (P less than 0.001). AECA levels correlated with low complement levels but not with anti-DNA antibodies to extractable nuclear antigens (ENA), anti-cardiolipin or anti-neutrophil cytoplasmic antibodies. The presence of AECA conferred a positive predictive value of 0.68 for the presence of nephritis. Twenty-five patients had active vasculitis at the time of assay and the highest AECA values were seen in patients with both nephritis and vasculitis. No correlation was seen with serum immunoglobulin levels and immune complexes did not bind significantly to the endothelial surface. The possible role of these antibodies as a marker in lupus nephritis is discussed.     

Endothelial cell binding by systemic lupus antibodies: functional properties and relationship with anti-DNA activity

Anti-DNA antibodies and anti-endothelial cell antibodies (AECA) are often detected in systemic lupus erythematosus (SLE). Anti-DNA antibodies can also bind the membrane of human umbilical vein endothelial cells (HUVEC), but little is known about the presence of AECA in the population of immunoglobulins from SLE sera that do not bind DNA. The aim of this study is to analyse the ability of anti-DNA and non-anti-DNA antibodies from SLE sera to bind endothelial cell antigens and to investigate their pathogenic potential. Both anti-DNA and non-anti-DNA antibodies display AECA activity by immunoprecipitation and flow cytometry and in some patients recognize antigens of identical molecular weight. Complement-dependent cytotoxicity on HUVEC was not detected with either anti-DNA or non-anti-DNA antibodies. Similarly, apoptosis was not induced in HUVEC and HL60 incubated with anti-DNA or non-anti-DNA antibodies, as shown by the DNA hypodiploid content. These data indicate that AECA are highly heterogeneous, as they recognize a wide variety of surface molecules on HUVEC and equally present in anti-DNA and non-anti-DNA antibodies from SLE patients.     

Anti-endothelial cell antibody in preeclampsia: clinical findings and serum cytotoxicity to endothelial cell.

BACKGROUND: The role of anti-endothelial cell antibody (AECA) in systemic vasculitis has been reported. One candidate which may disrupt vascular function is AECA. In order to investigate the role of AECA in preeclampsia, the incidence of AECA positive patients, the characteristics of the clinical findings of AECA positive patients and also the cytotoxicity of AECA positive serum for cultured endothelial cells was studied. METHODS: Serum samples were taken from 57 preeclampsia (including 37 severe cases) and 46 normal pregnant women. The AECA were measured by ELISA using human umbilical vein endothelial cells. The cytotoxicity for cultured endothelial cells by test serum was measured by using 51Cr release assay. RESULTS: The incidence of IgG and IgM AECA were revealed in 26.3% and 10.5% of preeclampsia respectively. AECA was detected more frequently in severe (29.7%) than in mild preeclampsia (20.0%). In cases with severe proteinuria of greater than 200 mg/dl we detected a significantly higher incidence of AECA than in mild cases (p < 0.04). The incidence of AECA was not significantly increased in cases with severe hypertension or IUGR. The AECA positive sera had greater cytotoxic activity on endothelial cells than AECA negative sera (p < 0.03). CONCLUSIONS: The appearance of AECA is related to the severity of proteinuria and the cytotoxicity to endothelial cells by AECA positive sera may play a role in causing the endothelial damage in preeclampsia.     

A Prospective Study on Anti–endothelial Cell Antibodies in Patients with Systemic Lupus Erythematosus

IgG anti–endothelial cell antibodies (AECA) were detected in 48.5% of patients with active systemic lupus erythematosus (SLE) and in 7% of patients during remission and were associated with the development of diffuse proliferative lupus nephritis. Sixteen AECA-positive patients were prospectively studied for 25.2 ± 2.9 months. Serial AECA levels correlated with disease activity in 10 (62.5%) patients. Seven (43.8%) of 16 patients remained AECA positive during clinical remission. Among four episodes of disease exacerbation and 16 instances of clinical improvement, 85% (17 episodes) were accompanied by corresponding changes in the level of AECA, while corresponding changes in C₃, anti–nuclear antibodies, and anti–double-stranded DNA antibodies were noted in 60, 60, and 80% of cases, respectively (p= not significant). AECA served as the only serologic marker of altered disease activity in five episodes, when C₃, ANA, and anti–double-stranded DNA levels remained unaltered. We conclude that the level of AECA can serve as a marker of disease activity in SLE and that serial monitoring of AECA can complement other serologic parameters in the management of patients.     

New target antigens for anti-endothelial cell antibodies

Numerous connective tissue diseases, such as systemic lupus erythematosus (SLE), and infectious states, such as leprosy, are characterized by early vascular endothelial cell (EC) damage. There is substantial interest in the role of anti-EC antibodies (AECA) in such an injury. Due to the diversity of AECA-associated conditions, these autoantibodies are likely to be heterogeneous, and, therefore, identification of their antigens (Ag) to be difficult. They may be classified into three groups: membrane components, ligand–receptor complexes and Ag derived from the blood and attached to the cell surface. New technologies have been developed to sort it out, such as expression libraries and two-dimensional electrophoresis. A handful of Ag have hitherto been recognized viz. heat-shock protein 60 in SLE and leprosy, or plasminogen activator inhibitor-1 in SLE and Wegener granulomatosis. In reality, most of the target Ag for AECA remain to be identified.     

Human monoclonal anti-endothelial cell IgG-derived from a systemic lupus erythematosus patient binds and activates human endothelium in vitro

Our objectives were to obtain monoclonal anti-endothelial cell antibodies (AECA) from systemic lupus erythematosus (SLE) patients, to characterize their antigen specificity, and their capability to induce a pro-inflammatory and pro-adhesive endothelial phenotype, and to investigate the mechanism of endothelial cell (EC) activation in vitro. Monoclonal IgG AECA were generated by hybridoma formation with human SLE B cells. Antigen specificity was characterized by immunoblotting with enriched cell membrane fractions, by cytofluorimetry and by cell solid-phase ELISA. Endothelial activation was evaluated by measuring increases in U937 cell adhesiveness, adhesion molecule (E-selectin and ICAM-1) expression and IL-6 production. In addition, mechanisms of endothelial activation were investigated by assessment of NF-kappaB by measuring the loss of its inhibitor I-kappaB. mAb E-3 bound live EC and recognized a 42 kDa EC membrane protein, it enhanced U937 adhesiveness, E-selectin and ICAM-1 expression and IL-6 production, and caused the loss of I-kappaB. We conclude this is the first in vitro demonstration that a human monoclonal AECA from a SLE patient reacts with a constitutive endothelial membrane antigen and induces a pro-inflammatory endothelial phenotype through NF-kappaB activation.