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.     

心肌肌凝蛋白轻链—1的免疫组化研究

为了研究心肌肌凝蛋白轻链－１（ＣＭＬＣ－１）组织特异性的特点，并对其构型在心肌发育过程中的表达规律以及不同哺乳动物ＣＭＬＣ－１构型的生化规律进行探讨，应用抗人心室肌ＣＭＬＣ－１，单克隆抗体（ＭｃＡＢ）对成人和不同胎龄胎儿的心肌和其它组织，不同种属哺乳动物的心肌进行了免疫组织化学研究。结果发现：成人心室肌，慢性骨骼肌均发生强烈反应，成人心房，胎儿心室肌发生交叉反应，而其它组织加快骨骼肌，平滑肌以及肌     

Avidity of technetium 99m glucarate for the necrotic myocardium: In vivo and in vitro assessment

Background  Similar to other^99mTc-based infarct-avid agents,^99mTc-glucarate localizes in myocardial infarcts. Whether severely ischemic viable myocytes sequester^99mTc-glucarate is uncertain. To assess the infarct specificity, in vitro and in vivo studies were performed. Methods and Results  H9C2 embryonic rat cardiocytes cultured under normoxia (N) or hypoxia (H) for 24 hours in 7.5 μCi^99mTc-glucarate were compared with necrotic cardiocytes. Mean H/N ratio (3.0±0.004, mean±SD) was significantly less than that of the necrotic/N ratio (39.9±6.5,p<0.01). Reperfused myocardial infarction (MI) in 4 dogs confirmed by²⁰¹Tl, (0.5 to 1.0 mCi) scintigraphy were imaged serially, with simultaneously injected mixture of^99mTc-glucarate and¹¹¹In-antimyosin Fab. Infarcts were detected scintigraphically within 4 to 10 minutes with^99mTc-glucarate.¹¹¹In-antimyosin required more than 1 hour. Myocardial distribution at 5 hours showed a direct correlation between^99mTc-glucarate and¹¹¹In-antimyosin uptake (r=0.99,p<0.0001). Both^99mTc-glucarate (r=−0.777,p<0.0001) and¹¹¹In-antimyosin (r=−0.775,p<0.0001) were inversely related to²⁰¹Tl distribution. Conclusions  The near perfect correlation between^99mTc-glucarate and¹¹¹In-antimyosin uptake (r=0.99) in reperfused canine MI and the insignificant glucarate uptake by viable cardiocytes in vitro attest to the avidity of^99mTc-glucarate for the necrotic myocardium and favor its use as a specific early marker of myocyte necrosis in acute MI. Supported in part by SBIR grant 1R43-HL54410-01 and Molecular Targeting Technology, Inc.     

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.     

EXPERIMENTAL STUDY OF RADIOIMMUOGUIDED SURGERY IN OVARIAN CARCINOMA

INTRODUCTIONThemaintreatmenttodateforovariancarcinomaissurgicalresection,thesocalledcytoreductivesurgery.Toperformacompleteresection,surgeonsusuallydependonvisualandtactilesensesaswellassurgicalinstinctswhicharerelativelycrudeinidentifyingsubclinicalmicroscopictumor.Radioimmuno--guidedsurgerysystem(RIGS)hasbeentakenoverthelast10yearsinbothlaboratoryandclinicalsettings.However,onlyafewreportswereseenaboutitsstudyinovariancancerworldwideespeciallyinourcountry.OurstudyincludeestablishingaR…     

2 132 例不同人群血清中SARS-CoV抗体水平调查

目的：探讨SARS-CoV(IgG)抗体检测辅助诊断非典型肺炎的临床应用价值。方法：采用ELISA方法及蛋白芯片法对2132例不同人群的血液标本检测SARS-CoV(IgG)抗体，分析SAILS康复者与其它疾病患者和正常人群抗体水平的差异。结果：用ELISA方法测定SARS康复者、肾病、恶性肿瘤、肺部疾病和糖尿病患者、其它疾病的SARS-CoV(IgG)抗体阳性率分别为75．6％(118／156)、29．3％(22／75)、22．3％(19／85)、22．7％(10／44)、13．6％(8／59)，500例健康查体人员全部为阴性，500例密切接触人员也为阴性。蛋白芯片方法测定SARS康复者的抗体阳性率为86％，对62例ELISA方法阳性的非SARS患者测定，仅发现4例阳性。结论：SARS-CoV(IgG)抗体可辅助回顾性诊断是否有SARS-CoV的感染。与ELISA方法相比，蛋白芯片方法具有更高的特异性和灵敏度。     

肝细胞生长因子对实验性急性胰腺炎细胞凋亡作用的研究

目的:观察外源性肝细胞生长因子(HGF)对小鼠轻症实验性急性胰腺炎的炎症反应及胰腺细胞凋亡的作用。方法:以雨蛙肽腹腔注射诱发小鼠轻症实验性急性胰腺炎。第1组以盐水腹腔注射作为对照,第2、3及4组每小时腹腔注射雨蛙肽诱发急性胰腺炎;第3组在造模前及造模中两次皮下注射重组人肝细胞生长因子(rhHGF);第4组同时皮下注射rhHGF及其单抗。观察HGF对炎症反应的干预作用及对腺泡细胞凋亡的影响。结果:炎症小鼠予每公斤体重4、20μgrhHGF时,与单纯炎症组相比,血清淀粉酶、脂肪酶下降,胰腺组织学炎症评分减少,胰腺腺泡细胞TUNEL凋亡指数增加,caspase3活性增强。予每公斤体重4或20μgrhHGF时,其效应无显著性差异。同时予rhHGF单抗的小鼠,上述指标与单纯炎症组相比差异无显著性。结论:HGF能减轻雨蛙肽诱导的小鼠轻症实验性急性胰腺炎。HGF单抗能消除HGF的生物学作用。HGF可以促进急性胰腺炎时腺泡细胞的凋亡,这可能是其保护胰腺的作用机制之一。