Bray Robert A, Tarsitani Christine, Gebel Howard M, Lee Jar-How
Department of Pathology, Emory University, Atlanta, Georgia, USA.
Methods Cell Biol. 2011;103:285-310. doi: 10.1016/B978-0-12-385493-3.00012-7.
For most solid organ and selected stem cell transplants, antibodies against mismatched HLA antigens can lead to early and late graft failure. In recognition of the clinical significance of these antibodies, HLA antibody identification is one of the most critical functions of histocompatibility laboratories. Early methods employed cumbersome and insensitive complement-dependent cytotoxicity assays with a visual read-out. A little over 20 years ago flow cytometry entered the realm of antibody detection with the introduction of the flow cytometric crossmatch. Cytometry's increased sensitivity and objectivity quickly earned it popularity as a preferred crossmatch method especially for sensitized recipients. Although a sensitive method, the flow crossmatch was criticized as being "too sensitive" as false positive reactions were a know drawback. In part, the shortcomings of the flow crossmatch were due to the lack of corresponding sensitive and specific HLA antibody screening assays. However, in the mid 1990s, solid phase assays, capable of utilizing standard flow cytometers, were developed. These assays used microparticles coated with purified HLA molecules. Hence, the era of solid-phase, microparticle technology for HLA antibody detection was born permitting the sensitive and specific detection of HLA antibody. It was now possible to provide better correlation between HLA antibody detection and the flow cytometric crossmatch. This flow-based technology was soon followed by adaptation to the Luminex platform permitting a mutltiplexed approach for the identification and characterization of HLA antibodies. It is hoped that these technologies will ultimately lead to the identification of parameters that best correlate with and/or predict transplant outcomes.
对于大多数实体器官移植和特定的干细胞移植而言，针对不匹配的人类白细胞抗原（HLA）抗原的抗体可导致移植器官早期和晚期功能衰竭。鉴于这些抗体的临床意义，HLA抗体鉴定是组织相容性实验室最关键的功能之一。早期的方法采用繁琐且不灵敏的补体依赖细胞毒性试验，并通过肉眼读取结果。20多年前，随着流式细胞术交叉配型的引入，流式细胞术进入了抗体检测领域。流式细胞术提高的灵敏度和客观性使其迅速成为一种首选的交叉配型方法，尤其对于致敏受者。尽管是一种灵敏的方法，但流式交叉配型因假阳性反应这一已知缺点而被批评为“过于灵敏”。部分原因在于，流式交叉配型的缺点是缺乏相应灵敏且特异的HLA抗体筛查试验。然而，在20世纪90年代中期，能够利用标准流式细胞仪的固相检测方法得以开发。这些检测方法使用包被有纯化HLA分子的微粒。因此，用于HLA抗体检测的固相微粒技术时代诞生了，它能够灵敏且特异地检测HLA抗体。现在有可能在HLA抗体检测和流式细胞术交叉配型之间提供更好的相关性。这种基于流式的技术随后很快被应用于Luminex平台，从而允许采用多重方法来鉴定和表征HLA抗体。人们希望这些技术最终能够确定与移植结果最相关和/或最能预测移植结果的参数。
