甲状腺自身抗体对常见甲状腺疾病诊治及预后估计的临床价值

目的 研究Graves病、原发甲减、亚甲炎、桥本病中TSH受体抗体(TRAb),甲状腺过氧化物酶抗体(TPOAb),甲状腺微粒体抗体(TMAb),甲状腺球蛋白抗体(TGAb)的变化趋势,从而揭示这四种甲状腺自身抗体对于以上四种甲状腺疾病诊治的意义.方法 测定68例上述甲状腺疾病患者的TRAb、TPOAb、TMAb、TGAb水平.结果 ①Graves病组TRAb较其他三种疾病增高更显著,并且治疗1年后随访,复发组该抗体滴度明显高于控制组;②桥本病组TPOAb明显高于其他3种甲状腺疾病,且治疗后1年组TPOAb水平与初发组无明显差异;③原发甲减TPOAb、TMAb、TGAb显著增高,提示可能由桥本病所致;④Graves病,原发甲减,桥本病TGAb水平较亚甲炎高,但在原发甲减与桥本病中无明显差异;⑤Graves病、原发甲减、桥本病TMAb水平较亚甲炎高,但在Graves病与原发甲减、Graves病与桥本病中无明显差异.结论 TRAb在对Graves病确诊、疗效及预后估计方面均具有重要意义,在Graves病复发后可再度增高.TPOAb对桥本病诊断治疗及预后估计方面的意义优于其他抗体,可视为桥本病的特异性抗体、Graves病、原发甲减、桥本病TMAb及TGAb水平较亚甲炎高,但在此三种疾病的鉴别诊断中无显著意义.     

甲状腺过氧化物酶抗体球蛋白抗体微粒体抗体在甲状腺自身免疫性疾病中的改变

目的 观察甲状腺过氧化物酶抗体(TPO Ab)、甲状腺球蛋白抗体(TG Ab)和甲状腺微粒体抗体(TM Ab)在自身免疫性甲状腺疾病(AITD)中的改变,探讨TPO Ab在临床诊断和治疗上的作用和意义.方法 收集AITD患者,根据甲状腺功能不同分为甲状腺功能亢进(简称甲亢)组(Graves病、GD,57例)、甲低组(桥本氏甲状腺炎、HT,48例)、亚甲低组(41例)和AITD复诊组(甲状腺功能恢复正常1～6个月,41例).另取一级亲属无GD或HT的健康人群53例为对照组.采用放射免疫分析法检测血清中甲状腺自身抗体(TPO Ab、TG Ab和TM Ab)及甲状腺激素和促甲状腺激素(FT3、FT4,sTSH)水平.结果 甲亢组、甲低组和亚甲低组中TPO Ab阳性率(87.70%、97.20%、100.00%)均明显高于同组内TG Ab阳性率(43.90%、60.42%、48.78%)和TM Ab阳性率(43.90%、79.10%、60.98%);3种甲状腺自身抗体的阳性率和阳性患者的抗体水平均高于相应的对照组.AITD复诊组的TPO Ab阳性患者的抗体水平[(683.04±606.55)kU/L]明显低于甲亢组、甲低组和亚甲低组[(1049.31±941.00)、(106 440.79±272.38)、(5133.01±4449.67)kU/L].结论 TPO Ab在AITD的诊断更具有代表意义,抗体水平为AITD治疗及预后评估提供了重要的依据.     

亚临床甲状腺功能异常临床细胞病理学分析

目的 探讨亚临床甲状腺功能减退症(亚临床甲减)和亚临床甲状腺功能亢进症(亚临床甲亢)的病因及临床特点.方法 对1994年9月至2005年12月北京大学第一医院符合诊断标准的90例亚临床甲减和48例亚临床甲亢患者的细针穿刺细胞病理学检查(FNAC)结果进行分析,同时测定甲状腺球蛋白抗体(TGAb)和甲状腺过氧化物酶抗体(TPOAb).结果 (1)根据细胞病理学诊断,亚临床甲减最多见于桥本甲状腺炎(81.71%),其次为甲状腺肿(12.19%),甲状腺腺瘤(3.66%)较少见;亚临床甲亢多见于桥本甲状腺炎(41.86%)、甲状腺肿(34.88%)、甲状腺腺瘤(13.95%).(2)亚临床甲减TGAb和TPOAb阳性率明显高于亚临床甲亢(P<0.01);桥本甲状腺炎TGAb和TPOAb阳性率在亚临床甲减中分别为83.08%和84.62%,在亚临床甲亢中分别为77.78%和55.56%.(3)桥本甲状腺炎的病理类型、相同病因的细胞病理形态在亚临床甲减和亚临床甲亢中未见明显差异.结论 桥本甲状腺炎是亚临床甲减的最常见病因,在亚临床甲亢的病因中也占有重要地位;亚临床甲减自身抗体阳性率明显高于亚临床甲亢;从细胞病理学上不能区分亚临床甲状腺疾病的功能状态.     

血清甲状腺自身抗体、碘摄入量与Graves病发病及临床转归的关系

目的探讨血清甲状腺刺激性抗体（TSAb）、甲状腺刺激阻断性抗体（TSBAb）等甲状腺自身抗体、碘摄入量与Graves病（GD）甲状腺功能亢进症（甲亢）发病和转归的关系。方法测定3个不同碘摄入量地区63例临床甲亢患者的血清TSAb、TSBAb、促甲状腺激素结合抑制免疫球蛋白（TBⅡ）、甲状腺过氧化物酶抗体（TPOAb）和甲状腺球蛋白抗体（TGAb）,2年后随访。TSAb和TSBAb采用转染了重组人促甲状腺素受体的中国仓鼠卵巢细胞（rhTSHR—CHO细胞）生物法测定。结果初访GD甲亢患者TSAb、TBⅡ和TSBAb的阳性率分别为80．9％、61．7％和6．4％,TSAb和TBⅡ任-抗体阳性率为91．5％,显著高于对照组。TSAb和TBⅡ的一致率为59．6％。GD甲亢患者TSAb与TBⅡ（r=0．407）、甲状腺球蛋白（r=0、301）、甲状腺体积（r=0．317）正相关。初访碘过量地区GD甲亢患者TSAb阳性率（91．7％）显著高于轻度碘缺乏地区（66．7％）,3个地区GD患者TBⅡ、TPOAb、TGAb阳性率、活性和甲状腺体积无统计学差异。随访时患者分为甲状腺功能恢复组（G1）和未恢复组（G2）。G1组TSAb活性和甲状腺体积显著下降。当TPOAb滴度显著增高,且随访时继续保持高滴度时,甲状腺功能不易恢复。此时TSAb对甲状腺功能的影响降为次要因素。结论TSAb对GD甲亢的诊断和判断临床转归的意义大于TBⅡ,联合应用二者能提高GD甲亢患者促甲状腺激素受体抗体的检出率;GD甲亢的转归与TSAb、TPOAb浓度和甲状腺体积有关。     

甲状腺淋巴细胞浸润与甲状腺自身抗体水平的相关性分析

目的探讨甲状腺淋巴细胞浸润与甲状腺自身抗体水平的关系。方法依据甲状腺针吸细胞学检查（FNAC）中淋巴细胞浸润程度由轻到重将249例甲状腺疾病患者分为5组,观察各组甲状腺球蛋白抗体（TGAb）、甲状腺微粒体抗体（TMAb）、促甲状腺激素受体抗体水平,分析淋巴细胞浸润程度与自身抗体阳性率的关系。结果甲状腺不同淋巴细胞浸润程度间TGAb、TMAb水平差异均有统计学意义（P均〈0.05）,TGAb、TMAb强阳性率（〉60%）与甲状腺淋巴细胞浸润程度存在正相关（r=1.00、0.90,P=0.00、0.04）。结论甲状腺自身抗体强阳性能较好地反映甲状腺淋巴细胞浸润情况,可作为FNAC的选择依据。     

多囊卵巢综合征患者血清甲状腺特异抗体检测及临床意义

目的探讨自身免疫性甲状腺炎（AIT）与多囊卵巢综合征（PCOS）的关系。。方法72例PCOS患者为观察组,68例健康体检的育龄期妇女为对照组。测定两组血清甲状腺特异抗体[包括甲状腺过氧化物酶抗体（TPOAb）、甲状腺球蛋白抗体（TGAb）]水平;比较观察组特异抗体阳性（指TPOAb及TGAb均阳性;诊断为AIT）与阴性（TPOAb及TGAb均阴性）者促卵泡素（FSH）／黄体生成素（LH）值及流产率。结果观察组及对照组特异抗体阳性率分别为23．6％、4．4％,P〈0．05。观察组特异抗体阳性者LH／FSH显著高于阴性者（P〈0．05）;特异抗体阳性者与阴性者流产率分别为23．5％、10．2％,但差异无统计学意义。结论PCOS患者AIT发病率显著高于正常人群,与雌、孕激素水平失衡有关;对PCOS患者应注意筛查甲状腺特异抗体,避免误诊误治。     

1759例甲状腺疾病患者TRAb，TGAb，TMAb联检结果分析

目的 通过对1759例甲状腺疾病患者TRAb,TGAb,TMAb联合检测结果的分析,探讨3种甲状腺自身抗体在甲状腺疾病鉴别诊断中的应用价值.方法 收集2002-2006年在山东省甲状腺疾病防治中心就诊的甲状腺疾病患者1759例,根据病史、症状、体征、化验及辅助检查结果分为6组.观察各组TRAb,TGAb,TMAb测定值的分布情况,各组间率的比较采用卡方检验.结果 (1)TRAb强阳性仅存在于甲亢患者(A、B两组),其他4组阳性率均很低,且测定值均小于50 u/L;(2)6组中TGAb,TMAb各档几乎均有分布,阳性率从高到低依次为D组(高达91.53%,94.92%)、C组、A组与B组、E组、F组;(3)A组与C组TGAb,TMAb强阳性率及TGAb阳性率差异均无统计学意义,仅TMAb阳性率c组略高于A组.结论 TRAb可作为甲状腺疾病的鉴别诊断依据;TGAb,TMAb强阳性对AITD及其他甲状腺疾病的鉴别有一定临床价值;不同类型甲状腺疾病患者的TGAb,TMAb存在异质性.     

Graves甲亢合并白细胞减少与甲状腺自身抗体相关性研究

目的对Graves甲亢合并白细胞减少患者的白细胞与甲状腺自身抗体[甲状腺过氧化酶抗体（TPOAb）、甲状腺球蛋白抗体（TGAb）]进行相关性分析,探讨TPOAb、TGAb与Graves甲亢合并白细胞减少的关系。方法采用放射免疫法检测20例Graves甲亢合并白细胞减少患者及20例白细胞正常的Graves甲亢患者的TPOAb、TGAb水平,并将白细胞数与TPOAb、TGAb水平进行相关性分析。结果 Graves甲亢合并白细胞减少患者的TPO-Ab、TGAb水平略高于白细胞正常的Graves甲亢患者,但无统计学差异。线性回归分析显示,Graves甲亢患者的白细胞数与TPOAb、TGAb之间没有相关性。结论 TPOAb、TGAb水平与Graves甲亢并白细胞减少无关。     

不同治疗手段及甲亢预后与甲状腺自身抗体关系的研究

目的探讨不同治疗手段及甲亢预后与甲状腺自身抗体之间的关系.方法采用放射免疫法(RIA)进行游离三碘甲状腺原氨酸(FT 3)、游离甲状腺素(FT4)、甲状腺球蛋白抗体(TGAb)、甲状腺抗过氧化物酶抗体(TPOAb)的测定;采用免疫放射法(IRA)进行超敏TSH(s-TSH)的测定;采用酶联免疫法(ELISA)进行TSH受体抗体(TRAb)的测定.结果未经治疗的无药组甲亢患者,其临床症状及体征最明显,甲状腺激素水平最高,TRAb阳性例数最多;抗甲状腺药物治疔组临床症状及体征有所缓解,但TRAb阳性例数较多、甲状腺激素水平较高;放射性碘(131Ⅰ)治疗后远期随访的甲亢患者,其临床症状及体征基本消失,甲状腺激素水平正常,预后最好,TRAb阳性例数为0,TGAb水平基本正常,TPOAb水平升高.结论抗甲药物、131Ⅰ治疗甲亢时,后者治疗后远期疗效最好,甲状腺自身抗体的变化最有利于患者的康复.     

自身免疫性甲状腺疾病和丙型肝炎

自身免疫性甲状腺疾病（AITD）和丙型肝炎均为常见病、多发病.近来丙型肝炎病毒（HCV）感染作为导致AITD发病的环境因素引起关注.一些研究发现甲状腺过氧化物酶抗体（TPOAb）和甲状腺球蛋白抗体（TgAb）阳性的患者或桥本甲状腺炎患者的抗-HCV抗体阳性率升高,慢性丙型肝炎患者的甲状腺功能减退症、TPOAb、TgAb的阳性率均增高.而另一些研究未发现丙型肝炎与AITD有关.     

输血前11606例患者HIV和HCV及TP抗体检测结果分析

目的为避免因输血传播疾病引起医疗纠纷和防止职业感染。方法采用酶联免疫吸附试验对2010-01～2011-03 11 606例就医患者进行输血前人类免疫缺陷病毒(HIV)抗体、丙型肝炎病毒(HCV)抗体、梅毒螺旋体(TP)抗体三项指标筛查。结果共检出阳性例数856例。结论患者输血前进行三项感染性指标检测,对诊断和预防医患交叉感染、减少医疗纠纷的发生具有重要意义。     

Pentameric complex of viral glycoprotein H is the primary target for potent neutralization by a human cytomegalovirus vaccine

Human cytomegalovirus (HCMV) can cause serious morbidity/mortality in transplant patients, and congenital HCMV infection can lead to birth defects. Developing an effective HCMV vaccine is a high medical priority. One of the challenges to the efforts has been our limited understanding of the viral antigens important for protective antibodies. Receptor-mediated viral entry to endothelial/epithelial cells requires a glycoprotein H (gH) complex comprising five viral proteins (gH, gL, UL128, UL130, and UL131). This gH complex is notably missing from HCMV laboratory strains as well as HCMV vaccines previously evaluated in the clinic. To support a unique vaccine concept based on the pentameric gH complex, we established a panel of 45 monoclonal antibodies (mAbs) from a rabbit immunized with an experimental vaccine virus in which the expression of the pentameric gH complex was restored. Over one-half (25 of 45) of the mAbs have neutralizing activity. Interestingly, affinity for an antibody to bind virions was not correlated with its ability to neutralize the virus. Genetic analysis of the 45 mAbs based on their heavy- and light-chain sequences identified at least 26 B-cell linage groups characterized by distinct binding or neutralizing properties. Moreover, neutralizing antibodies possessed longer complementarity-determining region 3 for both heavy and light chains than those with no neutralizing activity. Importantly, potent neutralizing mAbs reacted to the pentameric gH complex but not to gB. Thus, the pentameric gH complex is the primary target for antiviral antibodies by vaccination.Human cytomegalovirus (HCMV) is an important pathogen in transplant patients (1–5), and its infection can lead to invasive end-organ diseases, such as pneumonitis and hepatitis, as well as vascular pathology contributing to graft failure (4, 6, 7). HCMV is also the most common cause of in utero viral infections in North America and Europe, affecting 0.5–2% of newborns annually (8–10). Congenital HCMV infection can lead to symptomatic diseases at birth and also cause developmental disabilities in children (10, 11). Maternal seropositivity before conception protects against congenital transmission (12, 13), and both maternal humoral and cellular immunity are likely to contribute to the protection (14–16). Antibodies in particular are important for preventing congenital infection, serving as the first line of defense against maternal infection. It may also play a role in preventing transmission to the fetus, supported by the results of a small, nonrandomized study in pregnant women with primary HCMV infection, in which the passive immunity of monthly infusions of HCMV hyperimmune human IgG (HCMV-HIG) (200 mg/kg maternal weight) was ∼60% effective in protecting against congenital HCMV infection (17, 18). These studies suggest that it is feasible to develop a vaccine for preventing congenital HCMV infection and its sequelae. However, despite the fact that the Institute of Medicine has identified development of an effective vaccine for prevention of congenital HCMV as a top priority since 1999 (19), progress toward this goal has only been incremental (8, 20, 21). One of the hurdles to the efforts is our limited understanding of component of natural immunity associated with protection against HCMV infection.HCMV is a large, complex virus, with a genome capable of encoding >150 proteins (22–26). Because of the strict species specificity, options of animal models for HCMV research are limited (27). Thus, the functions of most HCMV antigens in viral infection in vivo and their roles as targets for host immunity are poorly understood. Furthermore, culture systems of single cell types have limitations for studying HCMV pathogenesis. Immunohistochemistry studies showed that HCMV can infect varieties of cells in vivo, including endothelial, epithelial cells, fibroblasts, and leukocytes (28–36). Many HCMV end-organ diseases, such as pneumonitis and gastroenteritis, are due to infection of the epithelial/endothelial cells in the affected organ (35–39). However, common laboratory strains, such as AD169 and Towne, were culture-adapted in fibroblast cells, with genomic mutations (22, 24, 40) and, more importantly, have lost their tropism to endothelial and epithelial cells, in contrast to pathogenic clinical isolates (32, 33, 41, 42).Loss of viral tropism to endothelial and epithelial cells was mapped to various mutations in the viral UL131-128 locus, and these mutations abrogated the expression of the pentameric glycoprotein H (gH) complex, composed of gH, gL, UL128, UL130, and UL131 proteins, a determinant for viral tropism to endothelial and epithelial cells (42–44). Because the pentameric gH complex is missing in common laboratory strains (42, 43), its importance in viral tropism, viral pathogenesis, and vaccine design was not fully appreciated until recently (42, 45). With this understanding, it is not surprising that Towne virus and recombinant glycoprotein B (gB) vaccines, although with ∼50% efficacy against primary infection in the clinic (46–49), induced poor neutralizing titers against viral infection of epithelial cells, in contrast to immune sera from HCMV-seropositive donors (50, 51). Thus, missing the pentameric gH complex is likely a deficiency in antigen composition for both vaccines (50). Studies of monoclonal antibodies (mAbs) isolated from HCMV-seropositive donors or polyclonal IgG enriched for antigen specificity supported the hypothesis that the pentameric gH complex, not gB, appears to be important for neutralizing activity in human subjects with natural infection (52).We recently described an experimental vaccine virus in which expression of the pentameric gH complex was restored (53). Unlike the parental AD169 virus and the recombinant gB vaccine, this virus can elicit high levels of neutralizing antibodies in rabbits and rhesus macaques (53). To support clinical development of this vaccine centered its concept on the pentameric gH complex, we established a comprehensive panel of 45 mAbs from a single rabbit that received vaccination. Of the 45 mAbs, 25 had neutralizing activity against viral entry in epithelial cells, including 11 elite neutralizers with ≥10-fold greater potency than HCMV-HIG. Biochemical analysis demonstrated that all elite neutralizers preferentially bound to the virus expressing the pentameric gH complex, and the majority of elite neutralizers (8 of 11) specifically recognized a recombinant form of the pentameric gH complex. Interestingly, binding affinity for intact virions was not correlated with neutralizing activity. Moreover, genetic analysis of the 45 mAbs based on their heavy- and light-chain sequences identified at least 26 B-cell linage groups characterized by distinct binding or neutralizing properties. In addition, neutralizing antibodies had longer complementarity-determining region 3 (CDR3) for both heavy and light chains than those of antibodies with no neutralizing activity. These data establish the importance of the pentameric gH complex as the primary target for potent neutralizing antibodies by vaccination, and support development of an experimental HCMV vaccine featuring the pentameric gH complex.     

Anti-endothelial cell antibodies to the endothelial hybridoma cell line (EAhy926) in systemic lupus erythematosus patients with antiphospholipid antibodies

The endothelial hybridoma (EAhy926) cell line was employed to clarify whether antiphospholipid antibodies (aPA) [lupus anticoagulant (LA), antiprothrombin antibody (aPT) and/or anticardiolipin antibody (aCL)] and anti-endothelial cell antibodies (AECA) are identical, and establish whether β₂-glycoprotein I (β₂-GPI) is needed for reactivity of aPA to endothelial cells. Ig-G AECA was positive in 9/30 SLE patients with aPA (30.0%) and 10/22 SLE patients negative for aPA (45.5%). Ig-M AECA was positive in one SLE patient with aPA and one SLE patient without aPA. AECA-positivity was not significantly different among unfixed, TNF-stimulated and fixed EAhy926. The influence of β₂-GPI on the reactivity of serum to EAhy926 was minimal, and absorption experiments of serum with cardiolipin-liposome/β₂-GPI or phosphatidylserine-liposome/prothrombin gave little evidence of cross-reactivity of aPA and AECA. The results of our study suggest that aPA and AECA may have partially cross-reacted, but were different antibodies. However, further study is needed to clarify the clinico-pathological significance of AECA.     

Determination of antiplatelet antibody activity in sera cytotoxic for human lymphocytes

Pooled serum aliquots obtained from sensitized potential renal allograft recipients on chronic hemodialysis were evaluated for their lymphocytotoxicity titers against the lymphocytes and then for alloantibodies against the platelets of 7 random donors by 5 methods. Platelet donor specific lymphocytotoxicity was present in 93% of 42 combinations. Of the positive combinations, 57% had a positive test for antiplatelet activity by the 14C serotonin release assay, 16% by the platelet aggregation method, and 19% as judged by acid phosphatase availability on the platelet membrane. No serum tested released beta-glucoronidase or lactic dehydrogenase. No correlation of the height of the titer of antiplatelet activity with that for lymphocytoxicity was detected. Thus, even in sera demonstrating significant activity against donor lymphocyte antigens, detection of associated platelet antibody activity is not uniform. Thus, a positive lymphocytoxic titer does not necessarily predict detectable antiplatelet activity. Therefore, additional tests for detection of antiplatelet activity should also be considered. This study shows that of the tests evaluated, the 14C serotonin release assay is the most sensitive for detection of antiplatelet antibodies.     

单克隆抗独特型抗体NP30用于日本血吸虫病血清学诊断的研究

本研究用单克隆抗独特型抗体NP30与日本血吸虫肠相关抗原(GAA)和可溶性虫卵抗原(SEA)检测了702份不同病期及正常人群中的血清抗体,结果显示,在急性感染时,NP30抗体的检出率为98％,与GAA(94％)和SEA(98％)的无差别。在慢性感染时NP30抗体的检出率为87％,与GAA(86％)的无差别,但低于SEA(98％)的。在正常人群中,上述3种的抗体假阳性率均为3％左右,无差别。NP30的抗体滴度几何均数在急性血吸虫感染时高于GAA的,低于SEA的,在慢性感染时低于后两者,提示NP30的抗体在血吸虫感染期间出现比较早,消退较快。上述结果提示,NP30可以替代虫源性抗原,用于日本血吸虫病诊断。     

From the Cover: Construction of a rationally designed antibody platform for sequencing-assisted selection

Antibody discovery platforms have become an important source of both therapeutic biomolecules and research reagents. Massively parallel DNA sequencing can be used to assist antibody selection by comprehensively monitoring libraries during selection, thus greatly expanding the power of these systems. We have therefore constructed a rationally designed, fully defined single-chain variable fragment (scFv) library and analysis platform optimized for analysis with short-read deep sequencing. Sequence-defined oligonucleotide libraries encoding three complementarity-determining regions (L3 from the light chain, H2 and H3 from the heavy chain) were synthesized on a programmable microarray and combinatorially cloned into a single scFv framework for molecular display. Our unique complementarity-determining region sequence design optimizes for protein binding by utilizing a hidden Markov model that was trained on all antibody-antigen cocrystal structures in the Protein Data Bank. The resultant ∼10¹²-member library was produced in ribosome-display format, and comprehensively analyzed over four rounds of antigen selections by multiplex paired-end Illumina sequencing. The hidden Markov model scFv library generated multiple binders against an emerging cancer antigen and is the basis for a next-generation antibody production platform.     

The influence of anti-endothelial/antiphospholipid antibodies on fibrin formation and lysis on endothelial cells

The prothrombotic mechanisms associated with antiphospholipid antibodies remain incompletely defined. Antibody binding to endothelial cells in vitro is a feature of antiphospholipid antibody-positive sera. We hypothesised that impairment of endothelium-dependent fibrinolysis by antiphospholipid/anti-endothelial antibodies is a contributory factor in the pathogenesis of thrombosis. We also aimed to confirm the displacement of annexin-V from endothelial cells and enhanced fibrin formation. Binding of immunoglobulin (Ig) from antiphospholipid antibody-positive sera to endothelial cells was examined using a cell-based enzyme-linked immunosorbent assay. Effects on fibrin formation and lysis were examined on cultured endothelial cell monolayers. Plasminogen activator inhibitor-1 (PAI-1) was assayed in supernatants. We confirmed antibody binding to endothelial cells. With four of 14 antiphospholipid antibody-positive sera there was some prolongation of fibrin clot lysis time, consistent with impairment of endothelial fibrinolytic activity. Secretion of PAI-1 was significantly correlated with clot lysis time on endothelial cell monolayers incubated with antiphospholipid/anti-endothelial antibody-positive sera, but not with control sera. IgG from antiphospholipid antibody-positive sera had little effect on endothelial cell surface annexin-V expression. We conclude that impaired endothelial fibrinolysis is a potential prothrombotic mechanism in subjects with antiphospholipid antibodies. We were unable to confirm enhanced displacement of annexin-V from endothelium by antiphospholipid antibodies.     

自身免疫性肝炎患者自身抗体的测定及意义

目的:探讨自身抗体测定对诊断自身免疫性肝炎的临床意义．方法:采用间接免疫荧光法(IIF)检测47例自身免疫性肝炎患者、158例非自身免疫性肝炎患者及40例健康体检者体内抗核抗体(ANA)、抗平滑肌抗体(SMA)、抗中性粒细胞胞质抗体(ANCA)、抗线粒体抗体(AMA)等自身抗体,ELISA法检测抗MPO抗体,并对结果进行回顾性分析．结果:ANA、SMA及ANCA检出率的比较,结果显示AIH中阳性率最高为SMA(66．0％, 31／47),而非AIH中则为6．3％(10／158),两组差异有非常显著性意义(P<0．01)．经X2检验, SMA、AMA和MPO抗体检测在AIH与PBC中,均有非常显著性意义(P<0．01)．AIH各型自身抗体检测结果表明,AIH-Ⅰ与ANA、SMA和ANCA相关,AIH-Ⅱ与LKM相关,而AIH=Ⅲ与SLA和ANCA相关．结论:血清自身抗体的检测对诊断、治疗和阻止自身免疫性肝炎的发展有着十分重要作用,对提高AIH在临床上同其他肝病鉴别诊断和治疗有着非常重要的意义．     

噬菌体抗体库的构建及人源抗内毒素类脂A抗体的筛选

目的寻找一种治疗内毒素血症及其并发症较为有效的途径.方法从感染革兰阴性杆菌J7患者体内含有内毒素类脂A抗体的淋巴细胞中提取mRNA,经反转录再从IgG重链Fd两端及轻链通用引物分别扩增Fab基因片段,依次插入到噬菌体载体pCOMR3中,电穿孔转入大肠杆菌XL-blue,经辅助病毒VCSM13感染,重组噬菌体溶源裂解.结果Fab抗体表达于噬菌体CPⅢ的N端,噬菌体抗体库的容量为4.8×106,筛选出抗内毒素类脂A抗体,经LPS蛋白对噬菌体抗体库进行3轮淘选,使抗内毒素类脂A的特异性抗体得到100倍的富集.结论通过直接和竞争ELISA实验筛选出3株结合活性好的特异性抗体,为下一步应用研究奠定了基础.