Prevalence and immunogenicity of HNA-3a and -3b antibodies and antigens

HNA-3a 和 -3b 抗体和抗原的流行率和免疫原性

• 批准号: 8031461
• 负责人: Richard Herbert Aster
• 金额: $ 24.98万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8031461
• 关键词: Acute Lung Injury; Address; Alleles; Alloantigen; Amino Acid Substitution; Antibodies; Antigens; Biological Assay; Biological Response Modifiers; Blood; Blood Transfusion; Blood donor; Blood donor screening; Cells; Complication; Detection; Donor Selection; Exposure to; FDA approved; Female; Genetic Polymorphism; Grant; HLA Antigens; Immunoassay; Incidence; Individual; Insecta; Isoantibodies; Laboratories; Length; Leukocytes; Membrane; Molecular; Mutate; Patients; Peptides; Phase; Phenotype; Population; Pregnancy; Prevalence; Property; Proteins; Reaction; Recombinants; Research; Research Personnel; Safety; Sampling; Serological; Serum; Solid; System; Testing; Time; Transfusion; Work; base; blood product; choline transporter; extracellular; immunogenicity; improved; male; mortality; neutrophil; prototype; theories

DESCRIPTION (provided by applicant): Transfusion-associated acute lung injury (TRALI), currently the leading cause of transfusion- associated fatality, is triggered by transfusion of blood products containing leukocyte antibodies and/or biological response modifiers to susceptible patients. Although many different leukocyte antibodies have been shown to induce TRALI, those specific for the leukocyte antigen HNA-3a are especially prone to cause very severe, and often fatal reactions. Unfortunately, it has not been possible to screen blood donors routinely for anti-HNA-3a because its molecular properties have for many years eluded investigators and because HNA-3a was thought to be neutrophil-specific, a cell difficult to use in serologic studies. Thus, very little is known about the prevalence of anti-HNA-3a in blood donors and the immunogenicity of this antigen is poorly understood. Even less is known about antibodies that recognize HNA-3b, the allele of HNA-3a, although they should, in theory, be as likely to cause TRALI as anti-HNA-3a. We recently showed that HNA-3a is carried on choline transporter- like protein 2 (CTL2) and that the HNA-3a/b polymorphism is almost certainly determined by an R>Q154 (R=HNA-3a, Q = HNA-3b) amino acid substitution in the first extracellular loop of the 10- membrane-spanning CTL2 protein. These findings suggest ways to develop a practical assay for detection of anti-HNA-3a and anti-3b suitable for routine donor screening. In this application, we propose to generate recombinant and synthetic CTL2 and CTL2 fragments containing R154 and Q154 and characterize their reactions against a panel of HNA-3-specific antibodies. Constructs found to be most sensitive and specific for anti-CTL2 detection will be produced in quantity, formatted into a prototype solid phase immunoassay, and used to screen 7,000 serum samples from transfused, non-transfused and parous males and females to determine the prevalence of anti-HNA-3a in these populations. The Q154 version of these constructs should provide comparable information about anti-HNA-3b. Findings made are expected to 1) define, for the first time, the prevalence of anti-HNA-3a and anti- HNA-3b in blood donor populations; 2) characterize the immunogenicity of HNA-3 and HNA-3b (likelihood of an antibody being induced upon exposure to antigen through transfusion or during pregnancy) and 3) establish a basis for determining whether routine screening of blood donors for anti-HNA-3a and anti-HNA-3b is warranted. PUBLIC HEALTH RELEVANCE: Transfusion-associated acute lung injury (TRALI) is a serious complication of blood transfusion and the most common cause of transfusion-related mortality. Transfusion of antibodies specific for white blood cells (leukocytes) is a major cause of TRALI. Antibodies against a leukocyte antigen designated HNA-3a are especially prone to cause severe, often fatal TRALI but, for technical reasons, it has not been possible to test blood donors to detect them. Recent findings made in our laboratory make it likely that this obstacle can be overcome and studies to accomplish this are described in this application.))

描述（由申请人提供）：输血相关的急性肺损伤（TRALI）目前是输血相关死亡的主要原因，是由含有白细胞抗体和/或生物反应改性剂的血液产物引发的，引发了与易感患者的输血。尽管已经显示出许多不同的白细胞抗体会诱导Trali，但针对白细胞抗原HNA-3A的特异性抗体尤其容易引起非常严重且通常是致命的反应。不幸的是，由于它的分子特性多年来一直避免了抗HNA-3A，因此不可能常规地筛选献血者，并且由于认为HNA-3A被认为是中性粒细胞特异性的，这是一种难以在血清学研究中使用的细胞。因此，关于供血者中抗HNA-3A的患病率很少，对这种抗原的免疫原性的了解很少。关于识别HNA-3B（HNA-3a等位基因）的抗体的了解，尽管理论上应该像抗HNA-3A一样引起Trali。我们最近表明，HNA-3A携带在胆碱转运蛋白类似于蛋白2（CTL2）上，并且几乎可以肯定的是，HNA-3A/B多态性由R> Q154（r = HNA-3A，Q = HNA-3B）氨基酸替换为10-经膜型膜型ctl2蛋白质的第一个外胞外循环中的氨基酸替代。这些发现提出了开发用于检测抗HNA-3A和适用于常规供体筛选的抗HNA-3A和抗3b的实用测定方法的方法。在此应用中，我们建议生成包含R154和Q154的重组和合成CTL2和CTL2片段，并表征它们针对HNA-3特异性抗体的反应。发现最敏感和特异性的抗CTL2检测的结构将以数量的形式产生，格式化为原型固相免疫测定法，并用于筛选来自输血，非转移和parous和parous的男性和女性的7,000种血清样品，以确定这些群体中抗HNA-3A的患病率。这些结构的Q154版本应提供有关抗HNA-3B的可比信息。预计提出的发现将是1）首次定义抗HNA-3A和抗HNA-3B的患病率； 2）表征HNA-3和HNA-3B的免疫原性（通过输血或怀孕期间暴露于抗原后诱导抗体的可能性）和3）建立一个基础，以确定抗HNA-3A和抗HNA-3B的常规筛查是否筛选献血者。 公共卫生相关性：输血相关的急性肺损伤（TRALI）是输血的严重并发症，也是输血相关死亡率的最常见原因。对白细胞（白细胞）特异性抗体的输血是Trali的主要原因。针对白细胞抗原指定HNA-3A的抗体特别容易引起严重的，通常是致命的Trali，但出于技术原因，不可能测试献血者来检测它们。我们实验室中最新的发现可能可以克服这一障碍，并在此应用中描述了这一障碍。））））