Taaning E
Department of Clinical Immunology, Glostrup Hospital.
Dan Med Bull. 1992 Aug;39(4):343-54.
The development of an enzyme linked immuno-sorbent assay (ELISA) for detection of platelet antibodies and platelet specific-antigens has been described. In the assay, intact platelets fixed to the bottom of microplates have been used as targets. After incubation with serum, the attached antibody has been detected by antiglobulin serum labelled with enzyme followed by assay of the enzyme reaction with its substrate. The clinical significance of platelet antibodies in alloimmune neonatal thrombocytopenia (AINT), post-transfusion purpura (PTP), and refractoriness to platelet transfusion therapy has been described. The clinical significance of autoantibodies in autoimmune thrombocytopenic purpura (AITP) and the detection of platelet surface glycoproteins by a modification of the platelet ELISA has also been described. In addition, a survey of the present state of knowledge of human platelet immunology, especially platelet-specific alloantigens and alloantibodies has been given. The studies of AINT and PTP have revealed a human immune response gene located in the MHC region: The susceptibility to anti-Zwa alloimmunization is strongly associated with the HLA-B8 and even more DR3 and DRw52a tissue types. In the studies of PTP, the first example of PTP due to anti-Zwb (only detectable in the platelet ELISA) has been described, and an association of the presence of anti-Zw antibodies of IgG3 subclasses (defined in the platelet ELISA using monoclonal mouse antibodies) and destruction of autologous platelets has been described. In an investigation of Ig classes and IgG subclasses of platelet associated Ig (PAIg) in children suffering from autoimmune thrombocytopenic purpura, no correlation was found between the immunochemical properties of the PAIg and the clinical course of the disease. However, a correlation between increased amounts of PAIgG3 and very low platelet counts was observed. A modification of the platelet ELISA using monoclonal antibodies against glycoproteins (GP) has been described. Using antibodies against GPIb and GPIIb/IIIa, it has been possible to diagnose Glanzmann's thrombasthenia (GT) and Bernard-Soulier syndrome (BSS). Especially in the diagnosis of thrombocytopenic patients suspected for BSS where platelet aggregation studies has been impossible due to low platelet counts, the detection of decreased amounts of surface GPIb in ELISA has been of great value. In conclusion, the platelet ELISA has been found simple and sensitive, and suitable for routine investigations of AINT, PTP, AITP, and refractoriness to platelet transfusion.
本文描述了一种用于检测血小板抗体和血小板特异性抗原的酶联免疫吸附测定法（ELISA）。在该测定法中，固定在微孔板底部的完整血小板被用作靶标。血清孵育后，通过用酶标记的抗球蛋白血清检测附着的抗体，然后用其底物测定酶反应。文中描述了血小板抗体在同种免疫新生儿血小板减少症（AINT）、输血后紫癜（PTP）以及血小板输血治疗无效中的临床意义。还介绍了自身抗体在自身免疫性血小板减少性紫癜（AITP）中的临床意义以及通过改良血小板ELISA检测血小板表面糖蛋白的方法。此外，还对人类血小板免疫学的现有知识状况进行了综述，特别是血小板特异性同种抗原和同种抗体。对AINT和PTP的研究揭示了位于MHC区域的人类免疫反应基因：抗Zwa同种免疫的易感性与HLA - B8密切相关，与DR3和DRw52a组织类型的相关性更强。在PTP的研究中，描述了首例由抗Zwb（仅在血小板ELISA中可检测到）引起的PTP，并报道了IgG3亚类抗Zw抗体（在使用单克隆小鼠抗体的血小板ELISA中定义）的存在与自体血小板破坏之间的关联。在对患有自身免疫性血小板减少性紫癜儿童的血小板相关Ig（PAIg）的Ig类别和IgG亚类进行的调查中，未发现PAIg的免疫化学性质与疾病临床进程之间存在相关性。然而，观察到PAIgG3量增加与极低血小板计数之间存在相关性。文中描述了一种使用针对糖蛋白（GP）的单克隆抗体改良的血小板ELISA。使用针对GPIb和GPIIb/IIIa的抗体，可以诊断Glanzmann血小板无力症（GT）和Bernard - Soulier综合征（BSS）。特别是在诊断疑似BSS的血小板减少患者时，由于血小板计数低无法进行血小板聚集研究，ELISA中检测到表面GPIb量减少具有重要价值。总之，血小板ELISA已被证明简单、灵敏，适用于对AINT、PTP、AITP以及血小板输血无效的常规检测。
