Evaluation of trophoblast HLA-G antigen with a specific monoclonal antibody

A monoclonal antibody to HLA-G has been generated by immunizing HLA-A2.1/human β²-microglobulin (β²m) double transgenic mice with murine L cells transfected with both human β²m and HLA-G. This monoclonal antibody, designated as G233, has been found not to cross-react with other HLA class I antigens when tested on numerous cell lines by flow cytometry. With immunohistology, all populations of extravillous trophoblast (cell columns, interstitial trophoblast, endovascular trophoblast, placental bed giant cells) were stained. An extensive range of adult and fetal tissues was also tested but none reacted with monoclonal antibody G233, including those previously reported to express HLA-G mRNA, indicating that the protein has a highly restricted distribution. Failure to detect HLA-G in the fetal thymus raises the question as to how T-cell tolerance to this antigen is induced. Immunoprecipitation of trophoblast surface proteins with monoclonal antibody G233 revealed a heavy chain of 39 kDa and a light chain of 12 kDa, indicating that HLA-G expressed on the surface of trophoblast is complexed with p2m. However, sequential immunoprecipitation with monoclonal antibody W6/32 followed by monoclonal antibody G233 continued to detect a residual band of 39 kDa, suggesting that trophoblast surface HLA-G may also occur as free heavy chains not associated with p2m. Immunoprecipitation followed by two dimensional gel electrophoresis showed that monoclonal antibody G233 recognizes several iso-forms of HLA-G from trophoblast similar to the characteristic spot array previously described for HLA-G. This monoclonal antibody G233 will be highly useful in future experiments to elucidate the function of HLA-G.     

化学发光免疫法检测粒细胞抗体IgG的研究

作者以ＡＢＥＩ－ＣｏＣｌ＿２－Ｈ＿２Ｏ＿２作为化学发光指示系统，用固相竞争法检测５８例白细胞减少症患者、４１例无白细胞减少症患者及３２例正常人等的粒细胞抗体ＩｇＧ。判断标准采用正常人血清化学发光强度指数法。实验结果表明，白细胞减少症患者血清中粒细胞抗体阳性率明显高于无白细胞减少症患者（Ｐ＜０．００５）。本法敏感性为４３．１％，特异性为９０．２％，准确性为６２．６％，是一种灵敏而准确的方法，能将粒细胞包被试管在室温下较长时间保存，以供临床监测粒细胞抗体，大大缩短检测时间。     

组织型纤溶酶原激活物双单抗夹心ELISA建立及临床应用

本文动用自制ｔ－ＰＡ单克隆抗体建立ｔ－ＰＡ：Ａａ夹心ＥＬＩＳＡ法。结合ｔ－ＰＡ：Ａ、ＰＡＩ：Ａ测定，对５０例正常人，８７例肝病，４７例冠心病和１９例深静脉血栓 成患者的血浆ｔ－ＰＡ：Ａｇ、ｔ－ＰＡ：Ａ、ＰＡＩ：Ａ水平进行了研究。     

Induction of type II collagen-specific antibody production in blood lymphocyte cultures of rhesus monkeys (Macaca mulatta) with collagen-induced arthritis using the immobilized native antigen.

Peripheral blood mononuclear cells (PBMC) from Rhesus monkeys previously immunized with bovine type II collagen to induce arthritis were cultured with the same antigen. Because the native protein is poorly soluble in culture medium a heating step is often used. The antigen in this form induced PBMC proliferation, but epitopes for the induction of antibody production and arthritis were lost. To keep the native protein intact it was coated on affigel beads. With the immobilized antigen specific antibody production could be induced.     

Plant cardiac glycosides and digoxin Fab antibody

The potential application of the Digoxin Fab antibody (Wellcome Digibind) in the clinical management of plant poisoning was investigated. The cardiac glycoside contents of various Australian plants were studied using immunoassay techniques. The cross-reactions of the Fab antibody and two digoxin assay antibodies against extracts of these plants were also studied. Results obtained indicated that the Digibind antibody cross-reacted with a wide range of glycosides contained in Australian plants and therefore could be of use in the treatment of life-threatening plant poisoning.     

同型半胱氨酸及抗磷脂抗体与深静脉血栓形成和复发关系的研究

目的通过检测深静脉血栓形成（DVT）患者血浆中的同型半胱氨酸（Hey）和抗磷脂抗体（APLA）水平，探讨DVT形成和复发的因素，并寻找DVT复发的预报因子。方法选取2001年1月至2003年4月期间哈尔滨医科大学附属第二医院血管外科收治的60例DVT患者，其中初发组和复发组各30例，另随机选取门诊就诊经证实无DVT的下肢静脉曲张或深静脉功能不全的30例患者作为对照组，同时选取复发DVT的一级亲属组30例。用荧光偏振免疫法测定Hey，用酶联免疫吸附试验（ELISA）检测APLA中的抗心磷脂抗体（AcLA）水平（IgG、IgM型）并测定狼疮抗体（LA）阳性率。结果初发与复发DVT组患者血浆中的Hey和ACLA-IgG及ACLA-IgM水平均高于对照组和复发DVT一级亲属组，差异有统计学意义（P〈0．01）。复发DVT的一级亲属组的血浆中的Hey水平高于对照组（P〈0．05）。各组的LA阳性率差异无统计学意义（P〉0．05）。结论DVT患者血浆中Hey和APLA均升高，Hey和APLA与深静脉血栓形成有密切关系。Hey升高与DVT复发有关，可能是DVT复发的预报因子。     

用酶联免疫吸附术观察向附睾注射鱼肝油酸钠后抗精子抗体的变化

本研究用酶联免疫吸附术观察8只家兔在附睾注射鱼肝油酸钠后血清及精浆中的抗精子抗体。血清抗精子抗体阳性率为37.5～50.0%,最早检出率为术后2个月,6～7月达高峰。精浆抗精子抗体阳性率为25%,最早检出率为术后4个月,且此时己达高峰。与输精管结扎资料相比,发现血清抗精子抗体检出率偏低,而精浆相应抗体偏高,表明向附睾注射鱼肝油酸钠引起的免疫反应有别于结扎,以生殖道局部为主。     

Acetylcholine receptor antibody in myasthenia gravis

Radioimmunoassay techniques were used to detect antibodies to the acetylcholine receptor (AAChR) in 164 patients with adult-onset myasthenia gravis. AAChR levels above 0.6 nM/l were considered pathological and were found in 67% of the patients with an average value of 58.99 +/- 125.02 nM/l (0.6-900.0). Correlation, with clinical functional status, the histopathological thymus alterations and the different therapeutics used did not disclose any statistically significant differences.     

重症肌无力患者泼尼松治疗前后免疫学指标的变化

探讨泼尼松治疗重症肌无力（MG）免疫学机制。对382例MG患者在漏尼松中剂量冲击，小剂量维持疗法治疗前后，检测酰胆碱受体抗炎（AchRab)，突触前膜抗体（PsMab)，单个核细胞亚群（PBMC），肿瘤坏死因子（TNF），可溶性白介素－2受体（SIL－2R），β2－微球蛋白（β2－m)，以及红细胞免疫功能的变化。结果表明：MG患者在泼尼松治疗前后多项免疫指标有显著性的变化。为泼尼松治疗MG提供了评定疗效的免疫学指标，进一步阐明了MG发病的免疫学机制。     

子宫内膜异位症患者血清中靶抗体的检测和应用

目的 :建立一种ELISA方法 ,以检测子宫内膜异位症患者血清中抗转铁蛋白抗体。　方法 :用转铁蛋白包被的间接ELISA方法检测血清中的抗转铁蛋白抗体。血清标本来自 96例正常体检女性 ,6 4例妊娠妇女 ,72例儿科女童 ,以及 2 4例子宫内膜异位症患者。　结果 :子宫内膜异位症患者血清中的抗转铁蛋白抗体阳性率远远高于对照组 ,特异性为 99.0 % ,敏感度为 95 .8% ,准确性为 99.2 %。　结论 :子宫内膜异位症患者血清抗转铁蛋白抗体阳性率明显高于对照组 ,抗转铁蛋白抗体检测可作为子宫内膜异位症的非创伤性诊断方法