Automated high-throughput detection anti-HLA antibodies

自动化高通量检测抗 HLA 抗体

• 批准号: 7156115
• 负责人: John D Roback
• 金额: $ 9.95万
• 依托单位: TRANSFUSION AND TRANSPLANTATION TECH
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7156115
• 关键词: antibody titering; biotechnology; blood donor; blood group incompatibility; blood transfusion; evaluation /testing; high throughput technology; histocompatibility antigens; histocompatibility typing; human tissue; immunohematology; immunomagnetic separation; leukocyte antigen typing; lung injury; method development; neutrophil; therapy adverse effect

DESCRIPTION (provided by applicant): This is an STTR Phase I application to fund proof of concept studies for a sensitive, accurate, and fully-automated blood screening assay to detect antibodies implicated in TRALI (Transfusion Related Acute Lung Injury). TRALI has become the leading cause of post-transfusion mortality in industrialized countries. Current evidence suggests that many (if not the majority) of TRALI cases are due to antibodies in the donated blood component that bind to allogeneic HLA class I and class II antigens, as well as human neutrophil antigens (HNA), on leukocytes of the recipient. Activated and/or agglutinated recipient leukocytes then lodge in the small capillaries of the lung, leading to severe pulmonary edema and hypoxemia. Because treatment is largely supportive (including intubation and mechanical ventilation), and mortality rates of 6-10% or higher have been reported, preventative strategies are needed. In this application, we provide proof of concept for an accurate and automated screening assay for TRALI antibodies: anti-HLA class I antibodies in patient plasma can be detected with excellent sensitivity and specificity using human HLA class I-coated beads and a fully-automated blood bank multianalyzer (3Ti-Aegis). This workstation, developed through previous STTR support, has already been shown to perform rapid, accurate, and cost- effective blood typing with walk-away automation. The present study will add to the roster of tests available on this instrument, and will demonstrate that testing for antibodies implicated in TRALI is possible with throughput compatible with blood center screening. The Specific Aims of this Phase I application are: (1) To develop and optimize a multi-specificity bead cocktail for simultaneous detection of antibodies against HLA class-I, class-II, and neutrophil (e.g., HNA-1abc, HNA-4a) antigens; and (2) To perform an in-house clinical trial comparing test sensitivity and accuracy with externally performed manual assays. Successful completion of this work will support a Phase II application, with the goals of performing multicenter clinical trials with the assay leading to FDA 510(k) approval. Implications and relevance to public health: While blood transfusions are life-saving interventions, they can be complicated by side effects. 1 potentially lethal adverse effect is TRALI, in which antibodies in the blood donor can produce lung injuries in the transfusion recipient. This application proposes to develop a test that could be used to screen blood donors for these antibodies, thus preventing TRALI and improving patient outcomes.

描述（由申请人提供）：这是I阶段的I阶段应用，用于为概念研究提供资金证明，以进行敏感，准确和完全自动化的血液筛查测定法，以检测与TRALI有关的抗体（输血相关的急性肺损伤）。特拉利（Trali）已成为工业化国家输血后死亡率的主要原因。当前的证据表明，TRALI病例的许多（如果不是大多数）是由于捐赠的血液成分中与同种异体HLA I和II类抗原结合的抗体，以及人类嗜中性粒细胞抗原（HNA），对受体的白氧细胞上。然后激活和/或凝集的受体白细胞在肺的小毛细血管中产生，导致严重的肺水肿和低氧血症。由于治疗在很大程度上是支持性的（包括插管和机械通气），并且已经报告了6-10％或更高的死亡率，因此需要进行预防策略。在此应用中，我们为TRALI抗体的准确和自动化筛选分析提供了概念证明：可以使用人类I型I类涂层的珠子和完全启发性的血液库多亚储物库（3ti-aegis）以极好的敏感性和特异性来检测患者血浆中的抗HLA I类抗体。该工作站通过以前的STTR支持开发，已经显示出具有步行自动化的快速，准确且具有成本效益的血液打字。本研究将增加该仪器上可用的测试名册，并证明与与血液中心筛查兼容的吞吐量可能与Trali有关的抗体进行测试。该阶段应用的具体目的是：（1）开发和优化多种特异性珠鸡尾酒，以同时检测针对HLA类-I类，II类和中性粒细胞（例如HNA-1ABC，HNA-4A）抗原的抗体； （2）进行内部临床试验，将测试灵敏度和精度与外部执行的手动分析进行比较。这项工作的成功完成将支持II期应用程序，其目标是进行多中心临床试验，并获得了FDA 510（k）批准的测定。对公共卫生的影响和相关性：输血是挽救生命的干预措施，但它们可能会因副作用而变得复杂。 1潜在的致命不利影响是TRALI，其中献血者中的抗体可以在输血接受者中产生肺部损伤。该应用建议开发一种可用于筛查这些抗体的献血者的测试，从而防止Trali并改善患者的结局。