On the association of the platelet-specific alloantigen, Pena, with glycoprotein IIIa. Evidence for heterogeneity of glycoprotein IIIa.
On the association of the platelet-specific alloantigen, Pena, with glycoprotein IIIa. Evidence for heterogeneity of glycoprotein IIIa.
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关于血小板特异性同种抗原 Pena 与糖蛋白 IIIa 的关联。
DOI：
10.1172/jci113250
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
Kunicki,TJ
中科院分区：
文献类型：
--
作者：
Furihata,K;Nugent,DJ;Bissonette,A;Aster,RH;Kunicki,TJ
新生儿同种免疫性血小板减少性紫癜与一种新的血小板特异性同种抗原Pena相关。我们现在提供的直接证据表明，Pena决定因子与糖蛋白（GP） IIIa相关，但它不同于定义PlA系统的表位。通过ELISA，使用GPIIb特异性单克隆抗体（Tab）和GPIIIa特异性单克隆抗体（AP3）从产生游离GPIIb和GPIIIa的条件下制备的血小板裂解液中捕获并保持抗原，anti-Pena与AP3持有的GPIIIa反应，而与Tab持有的GPIIb不反应。在另一种ELISA中，从pla1阳性和pla1阴性的血小板中纯化的GPIIIa单独用作抗原，anti-Pena与两种等位基因形式的GPIIIa反应。通过放射免疫沉淀，anti-Pena在未还原或还原条件下，在十二烷基硫酸钠-聚丙烯酰胺凝胶中沉淀出具有电泳特征的单个表面标记膜蛋白，与GPIIIa相同。通过荧光细胞测定，来自几个供体的血小板，无论PlA表型如何，结合的抗- pena量大致相当于PlA1纯合子（A1/A1）血小板结合的抗-PlA1量的一半，大致等于PlA1杂合子（A1/A2）血小板结合的抗-PlA1量。使用供体PlA1和Pena纯合子的血小板进行定量间接结合试验，在饱和状态下，每个血小板结合14- 24000分子的抗Pena和41- 51000分子的抗PlA1。无论PlA表型如何，Anti-Pena完全抑制adp诱导的pena阳性血小板聚集。这些结果表明，Pena行列式与GPIIIa相关，但与PlA不同。图片
Neonatal alloimmune thrombocytopenic purpura associated with a new platelet-specific alloantigen Pena has been reported. We now provide direct evidence that the Pena determinant is associated with glycoprotein (GP) IIIa, but that it is distinct from epitopes that define the PlA system. By ELISA wherein monoclonal antibodies specific for GPIIb (Tab) and specific for GPIIIa (AP3) were used to capture and hold antigens from a platelet lysate prepared under conditions that generate free GPIIb and GPIIIa, anti-Pena reacted with GPIIIa held by AP3 but not with GPIIb held by Tab. In an alternative ELISA where purified GPIIIa from both PlA1-positive and PlA1-negative platelets were used individually as antigen, anti-Pena reacted with both allelic forms of GPIIIa. By radioimmuno-precipitation, anti-Pena precipitated a single surface-labeled membrane protein with electrophoretic characteristics in sodium dodecyl sulfate-polyacrylamide gels, under nonreduced or reduced conditions, identical to those of GPIIIa. By fluorocytometry, platelets from several donors, regardless of PlA phenotype, bound an amount of anti-Pena roughly equivalent to one-half that amount of anti-PlA1 bound by PlA1 homozygous (A1/A1) platelets and roughly equal to that amount of anti-PlA1 bound by PlA1 heterozygous (A1/A2) platelets. Using platelets from donors typed homozygous for PlA1 and Pena in a quantitative indirect binding assay, 14-24,000 molecules of anti-Pena and 41-51,000 molecules of anti-PlA1 were bound per platelet at saturation. Anti-Pena completely inhibited ADP-induced aggregation of Pena-positive platelets, regardless of PlA phenotype. These results indicate that the Pena determinant is associated with GPIIIa but distinct from PlA.Images