Detection of bilayer phospholipid-binding antibodies using flow cytometry

被引:18
作者
Eschwege, V [1 ]
Laude, I [1 ]
Toti, F [1 ]
Pasquali, JL [1 ]
Freyssinet, JM [1 ]
机构
[1] UNIV STRASBOURG 1, FAC MED, INST HEMATOL & IMMUNOL, STRASBOURG, FRANCE
关键词
antiphospholipid antibodies; lipospheres; apolipoprotein H (beta(2) glycoprotein I);
D O I
10.1046/j.1365-2249.1996.00908.x
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Antiphospholipid antibodies (APL) are usually detected using solid-phase immunoassays, where cardiolipin is the most common capture antigen. Phosholipids are believed to adopt a monolayer organization when coated onto polystyrene after evaporation of the solvent. However, bilayer phospholipids are probably those evidenced as microparticles or cell fragments circulating in vivo under various pathological circumstances. The surface density of monolayer phospholipids on polystyrene is six times lower than that of bilayer phospholipids. In order to assess the influence of phospholipid organization on the detection of APL, we prepared glass microspheres coated with bilayer phospholipids (cardiolipin, phosphatidylcholine, cholesterol). Such lipospheres enabled us to study the binding of antibodies in 1:100 diluted plasma samples from patients with anticardiolipin antibodies of Ige isotype previously diagnosed by ELISA. Among the 39 plasma samples analysed by flow cytometry, 17 showed positive IgG binding to lipospheres. Only four additional samples became positive when adding 20 mu g/ml apolipoprotein H. The specificity of the binding was demonstrated by complete reversibility with 1.4 mu M annexin V and with a large excess of liposomes of the same composition. The absence of correlation between liposphere and ELISA results suggests that different subgroups of antibodies are detected depending on the method. The detection of APL using bilayer phospholipids is an original assay and may represent a more physiopathological approach to the specificity of APL.
引用
收藏
页码:171 / 175
页数:5
相关论文
共 21 条
[1]  
ABUAF N, 1994, ANN BIOL CLIN-PARIS, V52, P365
[2]   MEASUREMENT OF ANTIPHOSPHOLIPID ANTIBODIES BY ELISA USING BETA-2-GLYCOPROTEIN-I AS AN ANTIGEN [J].
ARVIEUX, J ;
ROUSSEL, B ;
JACOB, MC ;
COLOMB, MG .
JOURNAL OF IMMUNOLOGICAL METHODS, 1991, 143 (02) :223-229
[3]  
BEVERS EM, 1991, THROMB HAEMOSTASIS, V66, P629
[4]  
DACHARYPRIGENT J, 1993, BLOOD, V81, P2554
[5]   ACTIVATION OF HUMAN PROTEIN-C BY BLOOD-COAGULATION FACTOR XA IN THE PRESENCE OF ANIONIC PHOSPHOLIPIDS - ENHANCEMENT BY SULFATED POLYSACCHARIDES [J].
FREYSSINET, JM ;
WIESEL, ML ;
GRUNEBAUM, L ;
PEREILLO, JM ;
GAUCHY, J ;
SCHUHLER, S ;
FREUND, G ;
CAZENAVE, JP .
BIOCHEMICAL JOURNAL, 1989, 261 (02) :341-348
[6]   HUMAN PLACENTAL ANTICOAGULANT PROTEIN - ISOLATION AND CHARACTERIZATION [J].
FUNAKOSHI, T ;
HEIMARK, RL ;
HENDRICKSON, LE ;
MCMULLEN, BA ;
FUJIKAWA, K .
BIOCHEMISTRY, 1987, 26 (17) :5572-5578
[7]   ANTICARDIOLIPIN ANTIBODIES (ACA) DIRECTED NOT TO CARDIOLIPIN BUT TO A PLASMA-PROTEIN COFACTOR [J].
GALLI, M ;
COMFURIUS, P ;
MAASSEN, C ;
HEMKER, HC ;
DEBAETS, MH ;
VANBREDAVRIESMAN, PJC ;
BARBUI, T ;
ZWAAL, RFA ;
BEVERS, EM .
LANCET, 1990, 335 (8705) :1544-1547
[8]  
GILBERT GE, 1992, J BIOL CHEM, V267, P15861
[9]  
HARRIS EN, 1990, BRIT J HAEMATOL, V74, P1, DOI 10.1111/j.1365-2141.1990.tb02530.x
[10]   A PHOSPHOLIPID-BETA-2-GLYCOPROTEIN-I COMPLEX IS AN ANTIGEN FOR ANTICARDIOLIPIN ANTIBODIES OCCURRING IN AUTOIMMUNE-DISEASE BUT NOT WITH INFECTION [J].
HUNT, JE ;
MCNEIL, HP ;
MORGAN, GJ ;
CRAMERI, RM ;
KRILIS, SA .
LUPUS, 1992, 1 (02) :75-81