柯萨奇B组病毒IgM抗体检测方法的建立及在产科中的应用

应用间接免疫荧光法检测孕产妇血清中柯萨奇Ｂ组病毒ＩｇＭ抗体（ＣＢＶ－ＩｇＭ）并与ＥＬＩＳＡ法作比较，两法结果无显著性差异（ｘ＾２＝０．５，Ｐ〉０．５），１３７０名早，中，晚期孕产妇血清ＣＢＶ－ＩｇＭ抗体检出率均高于非孕组（Ｐ〈０．０５）。     

抗心磷脂抗体与反复自然流产关系的探讨

应用间接ＥＬＩＳＡ法检测１０５例反复自然流产患者血清中的抗心磷脂抗体，同时对ＡＮＡ、扩ｄｓ－ＤＮＡ、抗－Ｓｍ，抗－ＲＮＰ、ＲＦ进行了测定。结果表明：ＲＳＡ组ＡＣＡ阳性率为２８．６％，明显高于其他自身抗体及对照组（Ｐ＜０．０１），ＡＣＡ与ＲＳＡ患者的孕龄及流产次数无相关性（Ｐ＞０．０５），三种类别Ｉｇ中，以ＩｇＧ、ＩｇＧ、ＩｇＭ型ＡＣＡ检出率较高，中等或高水平的阳性结果占３３．３％（１０／３０），其     

两种血清型HFRS患者病程中及病后4种特异性抗体测定分析

为评价汉滩及汉城型肾综合征出血热患者免疫状况，用ＥＬＩＳＡ法检测了５４例病程中，４７例病后１－６年ＨＦＲＳ患者（汉滩２４例，汉城２３例）血清中特异性ＩｇＡ、ＩｇＥ、ＩｇＧ、ＩｇＭ抗体。在病后血清中检出率及抗体滴度几何均数（ＧＭＴ）依次为ＨＦＲＳ－ＩｇＧ、ＩｇＡ、ＩｇＥ、ＩｇＭ，各本间差异显著（Ｐ〈０．００５），其中ＨＦＲＳ－ＩｇＧ抗体水平在病后第４年仍达１：１００以上（ＧＭＴ值）；ＨＦＲＳ－ＩｇＡ     

不孕、流产患者血清中三种Ig类抗卵巢自身抗体的研究

用亲和层析法纯化人卵巢抗原、建立ＥＬＩＳＡ间接法检测不孕、流产患者血清中三种Ｉｇ类抗卵巢抗体。结果表明：不孕、流产患者血清中ＩｇＧ、ＩｇＡ、ＩｇＭ三类抗卵巢抗体的水平与阳性率，均明显高于对照组（Ｐ＜０．０１）。提示此类抗体的检测，有助于了解不孕、流产的病因。     

血清中三种Ig类抗卵巢自身抗体的测定及其临床意义

用亲和层析柱纯化人卵巢抗原，建立ＥＬＩＳＡ间接法检测不孕、流产患者血清中三种Ｉｇ类抗卵巢抗体。结果表明：以健康孕妇（ｎ＝７０）Ａ值的Ｘ＋２ｓ为正常上限，不孕组患者血清中ＩｇＧ、ＩｇＡ、ＩｇＭ类抗卵巢抗体的阳性率分别为２２．７％（４８／２１１）、１４．２％（３０／２１１）和１２．３％（２６／２１１）；流产组患者血清中ＩｇＧ、ＩＧＡ、ＩｇＭ类抗卵巢抗体的阳性率分别为２０．６％（２２／１０７）、１５％（１６／１０７）和１２．１％（１３／１０７），均明显高于健康孕妇组（Ｐ＜０．０１）。不孕组以单独ＩｇＧ类或ＩｇＡ类抗体多见，流产组以ＩｇＧ、ＩｇＡ两类抗体为主。     

HEP-2细胞为靶细胞的ELISA检测血清抗核抗体

以ＨＥＰ－２细胞株为靶细胞包被塑料微孔扳，运用ＨＲＰ羊抗入ＩｇＧ为二抗对８例活动期ＳＬＥ，１例混合结缔组织病（ＭＣＴＤ），８例类风湿性关节炎，７例风湿性关节炎和３０例健康者做了血清抗核抗体的的ＥＬＩＳＡ检测，同时以间接免疫荧光法作对照。并对高滴度阳性血清作了稀释度与吸光度值的直线回归分析。结果表明，ＥＬＩＳＡ检测血清抗核抗体特异性为９８．５％，敏感性为１００％，值得推广应用     

抗β2GP1抗体检测方法的建立及临床意义

目的建立抗β２ＧＰ１抗体检测的ＥＬＩＳＡ方法。方法用亲和层析得到的纯化β２ＧＰ１为抗原。结果和结论１１２例被检血清抗β２ＧＰ１及抗心磷脂抗体的检测结果表明，两种方法的检测结果呈正相关（ＩｇＧ类ｒ＝０．６６７，ＩｇＭ类ｒ＝０．５５３）。抗β２ＧＰ１阳性组较阴性组患者血栓／多发性流产的发生率高，抗β２ＧＰ１的检测对血栓／多发性流产的发生有预示价值。为临床建立敏感、特异的诊断方法奠定了基础。     

解脲脲原体与新生儿感染

目的确定解脲脲原体（Ｕｕ）与新生儿肺炎等感染的关系。方法用间接血凝试验（ＩＨＡ）检测妊娠晚期孕妇血清Ｕｕ抗体，并对胎盘做Ｕｕ分离培养，用酶联免疫吸附试验（ＥＬＩＳＡ）检测部分胎儿脐血清Ｕｕ－ＩｇＭ抗体。并对孕妇血清Ｕｕ抗体阳性组和阴性组新生儿感染情况进行比较。结果１６０名孕妇血清中，８１例Ｕｕ抗体阳性，从其中１３例的胎盘中分离培养出Ｕｕ。用ＥＬＩＳＡ法检测５例胎盘Ｕｕ阳性的新生儿脐血Ｕｕ－ＩｇＭ，３例阳性。孕妇血清抗体阳性组的新生儿肺炎１６例，发热者１０例，两者总发病率为５４．５％，较阴性组高（３７．９％，Ｕ检验Ｐ＜０．０５）。结论Ｕｕ是引起新生儿感染，特别是新生儿肺炎的主要病原体之一     

过敏性紫癜与T、B细胞的相关性探讨

目的：探讨过敏性紫癜与Ｔ细胞和Ｂ细胞免疫的关系。方法：对４２例过敏性紫癜患儿采用单克隆抗体免疫荧光法,进行了外周血Ｔ细胞亚群检测。血清免疫球蛋白ＩｇＧ,ＩｇＡ,ＩｇＭ的测定采用琼脂单相免疫比浊法,可溶性白介素２受体（ＳＩＬ－２Ｒ）测定采用双抗体夹心（ＥＬＩＳＡ０法,同时做了３０例对照。结果：ＣＤ３,ＣＤ４,ＣＤ８活性值均低于正常对照组,ＣＤ３（ｔ＝１０．１８,Ｐ〈０．０１）,ＣＤ４（ｔ＝７．６６,     

固相间接血凝法检测伤塞杆菌O抗体及其临床初步应用

建立了固相间接血凝法（ＳＰ－ＰＨＡ）用于检测伤寒杆菌Ｏ抗并与间接血凝法（ＬＰＳ－ＰＨＡ），酶联ＡＳ蛋白ＥＬＩＳＡ法及经典的肥达氏试验作了比较，ＳＰ－ＰＨＡ法敏感性优于ＬＰＳ－ＰＨＡ《肥达氏试验（Ｗｉｄａｌ‘ｓ）与酶联Ａ蛋白ＥＬＩＳＡ法（ＳＰＡ－ＥＬＩＳＡ）相近。伤寒患者血清Ｏ抗全阳性纺为８８．２％（１５／１７），发热待查阳性纺为３．６％（２／５５），３０例献血员血清Ｏ抗体均为阴性。该法敏感、特异、     

血管内皮细胞损伤性疾病患者抗内皮细胞抗体和抗心磷脂抗体检测及其意义

目的探讨抗内皮细胞抗体(AECA)及抗心磷脂抗体(ACA)与血管内皮细胞损伤性疾病的关系及意义。方法建立ELISA法检测AECA,采用ELISA法测定ACA,分别检测168例血管内皮细胞损伤性疾病患者中2种抗体的水平。结果168例血管内皮细胞损伤性疾病中冠心病、特发性血小板减少性紫癜、溶血性尿毒综合征及妊娠高血压综合征(妊高征)患者的AECA和ACA的阳性率分别为32.9%、22.7%、22.2%、38.9%和38.2%、31.8%、33.3%、44.4%,均显著高于正常对照组。结论AECA和ACA与血管内皮细胞损伤性疾病有一定的相关性。AECA与ACA无相关性。     

IgG antiendothelial cell autoantibodies from scleroderma patients induce leukocyte adhesion to human vascular endothelial cells in vitro. Induction of adhesion molecule expression and involvement of endothelium-derived cytokines.

IgG autoantibodies that bind human endothelial cells (AECA) were detected by ELISA in 30 of 42 samples of sera from patients with scleroderma. Pretreatment of human umbilical vein endothelial cells with AECA-positive scleroderma sera, or IgG purified from these sera, led to a dose- and time-dependent increase in the ability of the cells to bind human U937 monocytic cells. Threshold-active IgG concentrations were 1-10 micrograms/ml; effects were significant after 3 h and maximal after 6-12 h. IgG from AECA-negative sera or normal sera were without effect. Increased adhesion of U937 cells was accompanied by increased expression of endothelial intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin. Transfer of endothelial cell-conditioned media after pretreatment with AECA and immunodepletion of IgG demonstrated the presence of transferable activity that mimicked the effects of AECA. Treatment with neutralizing anticytokine antibodies indicated that IL-1, generated by the endothelial cells in response to AECA, was involved in the upregulation of adhesion molecules and U937 cell adhesion. We conclude that AECA can play a pathogenic role in scleroderma by activating endothelial cells, in part due to autocrine or paracrine actions of IL-1.     

急性脑梗塞患者AECA、TM检测的临床意义

目的:通过对急性脑梗塞患者血清抗血管内皮细胞抗体(Anti endothelial cell antibodies,AECA)和血浆可溶性血栓调节蛋白(soluble thrombomdulin,sTM)的联合检测,旨在探讨急性脑梗塞临床发病与血管内皮细胞损伤的内在关系.方法:采用间接荧光免疫法和双抗体夹心ELISA法检测66例急性脑梗塞患者(急性脑梗塞组)和35例献血员标本(对照组)的AECA、sTM水平. 结果:急性脑梗塞组的AECA阳性率显著高于对照组(P＜0.01);急性脑梗塞组治疗前sTM水平显著高于对照组(P＜0.01);急性脑梗塞组治疗3～4周后sTM水平与对照组相比无显著性差异(P＞0.05).结论:AECA、TM的检测可作为急性脑梗塞血管内皮损伤的观察指标,有助于脑血管病变的诊断和治疗.     

抗内皮细胞抗体和抗整合素αvβ3抗体测定在ITP与SLE鉴别诊断中的意义

目的探讨抗内皮细胞抗体（AECA）和抗整合素αvβ3抗体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EL1SA法分别测定40例ITP患者,43例SLE患者及44名正常人血清中的AECA和抗整合素αvβ3抗体水平。结果SLE组、ITP组患者血清AECA、抗整合素αvβ3抗体水平明显高于正常对照组（P〈0．01）SLE组患者血清AECA、抗整合素αvβ3抗体水平及阳性率显著高于ITP组（P〈0．01）。结论血清AECA和抗整合素αvβ3抗体的测定在鉴别诊断ITP和SLE中有显著的临床意义。     

Anti-endothelial cell antibodies: detection and characterization using a cellular enzyme-linked immunosorbent assay

Anti-endothelial cell antibodies (AECA) have been detected in autoimmune diseases such as systemic lupus erythematosus (SLE) and scleroderma (PSS) but their role in pathogenesis is unknown. Immunofluorescence, immunohistochemistry, complement-dependent antibody lysis, and radioimmunoassay have been used in the past to detect AECA. We have developed a rapid, sensitive, and quantitative cellular enzyme-linked immunosorbent assay (ELISA) to detect and characterize AECA. Sera were obtained from 28 normal volunteers, 28 patients with SLE, and 14 patients with PSS. We also performed studies in 47 patients with various monoclonal gammopathies. Endothelial cells (EC) were obtained from human umbilical veins by standard methods and subcultured on 96-well tissue culture plates without fixation. EC were then sequentially incubated with sera, peroxidase-conjugated goat anti-human Ig (IgG, IgM, or IgA), and substrate. Optical density readings were converted to arbitrary units by developing a standard curve. Heavy-chain specific antibodies were used to determine the class of AECA binding to EC. IgG was purified by using protein A columns and digested with pepsin to obtain F(ab')2 fragments. The mean units of AECA from normals were 19.3 for IgG and 12.5 for IgM. SLE sera showed significant levels of IgM AECA (37 units, P less than 0.001) but not IgG (29 units, P less than 0.1). PSS sera showed significant levels of both IgM AECA (38 units, P = 0.001) and IgG AECA (42.7 units, P less than 0.005). IgA AECA were not detected in normal, SLE, or PSS sera. Blocking Fc receptors with rabbit IgG did not affect the titer of IgG or IgM AECA.(ABSTRACT TRUNCATED AT 250 WORDS)     

冠心病患者血清中相关自身抗体的检测及临床意义

目的：探讨血清中抗氧化型低密度脂蛋白(ox-LDL)抗体和抗内皮细胞抗体(AECA)这2种自身抗体与冠状动脉粥样硬化的关系。方法：分别用酶联免疫吸附法(ELISA)和间接免疫荧光法(IIF)检测118例冠状动脉硬化性心脏病患及44名健康对照血清ox-LDL和AECA水平，同时用生化方法检测其血脂水平。结果：冠心病组血清抗ox-LDL抗体阳性率为32．20％，明显高于正常对照组阳性率6．82％．(P=-0．001)；血清抗内皮细胞抗体阳性率冠心病组为38．14％，健康对照组为13．64%(P=0．002)。在血脂正常的冠心病组中抗ox-LDL抗体阳性率25．00％也高于正常对照组(P=0．045)。结论：血清中上述2种自身抗体阳性率与冠状动脉粥样硬化密切相关，而且抗ox-LDL抗体阳性率与血脂无明显相关性，自身免疫反应是动脉粥样硬化的一种可能的发病机制。     

Pathogenic role of anti-endothelial cell antibodies in systemic vasculitis

Antiendothelial cell antibodies (AECA), a heterogeneous group of antibodies quite distinct from the ANCA family, have been detected in variety of diseases which share a varying degree of vessel wall damage. This review is mainly focused on Wegener's granulomatosis, Takayasu's arteritis and Kawasaki syndrome, which provide the best examples to evaluate the pathogenic and prognostic value of AECA. There is increasing evidence to show that AECA might be pathogenic in inducing autoimmune vascular disease. It is relevant to note that the presence and titre of AECA has been correlated with disease activity in systemic vasculitis. Experimental in vitro and in vivo models support a potential pathogenic role for AECA in sustaining immune-mediated vessel inflammation. Rather than being cytotoxic to endothelial cells, AECA are able to up-regulate the expression of adhesion molecules (E-selectin, intercellular adhesion molecule-1 and vascular cell adhesion molecule-1) and to induce the secretion of cytokine and chemokine which, in turn, cause leukocyte recruitment and adhesion. A recent idiotypic animal model has provided further evidence that AECA can be pathogenic.     

血清抗内皮细胞抗体在突发性耳聋诊断和预后中的价值

目的:检测突发性耳聋(sudden hearing loss,SHL)患者血清抗内皮细胞抗体(anti-endothelial cell antibody,AECA)的水平并探讨其临床价值。方法:采用酶联免疫吸附方法检测67例SHL患者(SHL组)及30例健康者(对照组)的血清AECA水平。分析血清AECA与SHL患者性别、年龄、听力损失程度、听力下降类型及预后之间的关系。结果:AECA阳性率在SHL组、对照组中分别为70.15%(47/67)、23.33%(7/30),差异有统计学意义(P=0.0000)。SHL组血清AECA浓度显著高于对照组(P=0.0001)。血清AECA阳性分布与患者性别、年龄、听力下降类型无显著相关,但与听力下降程度和预后呈显著相关(P=0.0366;P=0.0324);且在41.031ng/mL～162.26ng/mL范围内时,AECA水平与患者预后呈正相关(r=0.7346,P=0.0000)。对照组和SHL组ROC曲线下面积为0.7657[标准误为0.0479,95%可信区间为(0.66830～0.85594)]。结论:血清中的AECA可能在SHL的发病机制中发挥了重要作用,AECA还有望作为患者筛选及指导预后的血清学指标。     

类风湿关节炎患者AECA、VEGF、IL-17的检测及临床相关性研究

目的通过对类风湿关节炎(rheumatoid arthritis,RA)患者血清抗血管内皮细胞抗体(Anti endothelial cell anti-bodies,AECA)、血浆血管内皮生长因子(Vascular endothelial growth factor,VEGF)和白细胞介素-17(interleukin-17,IL-17)的检测,以探讨AECA、VEGFI、L-17在RA患者发病、病情进展中的相关性及其内在联系。方法采用间接免疫荧光法(IIF)和双抗体夹心ELISA法检测86例RA、45例骨关节炎(OA)、30例健康对照AECA的阳性率和VEGFI、L-17水平,VEGFI、L-17水平与红细胞沉降率(ESR)、超敏C反应蛋白(hs-CRP)、类风湿因子(RF)等指标进行相关性分析。结果 RA患者AECA阳性率为8.1%,高于OA患者的阳性率2.2%和健康对对照的阳性率0%(P0.05);RA活动期AECA阳性率为16.7%,高于RA缓解期的阳性率3.6%(P0.05);RA患者血浆IL-17和VEGF水平显著高于OA组和健康对照组(P0.05);RA患者活动期血浆IL-17和VEGF水平明显高于RA缓解组及健康对照组(P0.01);RA患者AE-CA滴度、血浆IL-17、VEGF水平与ESR、hs-CRP、RF的指标呈正相关。结论 AECA、VEGFI、L-17三者与RA的发病、病情活动存在一定的关系,IL-17、VEGF水平变化及AECA的滴度可作为临床观察RA病情活动、判断疗效及预后等方面的参考指标。     

抗内皮细胞抗体的检测及临床应用

目的 建立可以常规应用的免疫荧光法检测血清中抗内皮细胞抗体（AECA）。方法 对试验条件进行研究并与ELISA法对比，检测冠心病、脑梗死、肺疾患及脑血管障碍患者310例，青年健康组50例，老年健康组30例。结果 两法检测阳性率无显著差异（P＞0．05），符合率89．7％；试验表明，用两种方法检测冠心病组、脑梗死组的AECA阳性率均明显高于健康组（P＜0．01），肺疾患组与脑血管障碍组也高于健康组（P＜0．05），内皮素检测证实AECA阳性者与内皮素升高明显正相关。结论 自身免疫形成的AECA在促进内皮细胞损伤中具有重要意义；免疫荧光法快速、简便，适用于临床应用。