Development of novel reagents to identify xenograft reactive B cells

开发新试剂来鉴定异种移植反应性 B 细胞

基本信息

批准号：
10592419
负责人：
A Joseph Tector
金额：
$ 43.11万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10592419
关键词：
Affect Animal Organ Antibodies Antibody-Producing Cells Antigens Appearance Award B cell therapy B-Lymphocyte Subsets B-Lymphocytes Binding Blood Transfusion CRISPR/Cas technology Carbohydrates Cell Line Cross Reactions Data Development Family suidae Frequencies Genes Goals Grant HLA Antigens Heterophile Antigens Histocompatibility Homologous Gene Human Human Biology Immune response Immunity Immunoglobulin G Immunoglobulin M Immunoglobulins Individual Infection Knock-out Knowledge Major Histocompatibility Complex Malignant Neoplasms Molecular Mimicry Organ Organ Donor Organ Transplantation Patients Peripheral Blood Mononuclear Cell Persons Polysaccharides Population Predisposition Pregnancy Protein Array Proteins Rapid screening Reaction Reagent Recombinants Research Research Personnel Residual state Role Sampling Serum Solid Sorting Source Stains Testing Tissues Transplant Recipients Transplantation Waiting Lists Work Xenograft procedure allotransplant comparative cross reactivity detection sensitivity differential expression high throughput analysis high throughput screening human tissue improved innovation natural antibodies novel prevent protein complex replacement tissue rituximab screening success technology development tool

项目摘要

Project 3: Development of Novel Reagents to Identify Xenograft Reactive B Cells Project Summary A shortage of available donor organs is the most critical challenge facing organ transplantation today. Pigs are considered a promising source of replacement tissues. Unfortunately, xenotransplantation, the sharing of organs across species, is not clinically applied because humans exert a strong humoral immune response toward pig tissues. Our long-term goal has been to make pig tissues suitable for use in humans by preventing the binding of human antibodies. The central hypothesis of this project is that pig Major Histocompatibility Complex (MHC) proteins, known as Swine Leukocyte Antigens (SLA) in the pig, contribute to xenoantigenicity. We have shown that patients having antibodies against Human Leukocyte Antigens (HLA) often cross-react with the homologous SLA. We have also found that approximately 25% of the population who lack HLA antibodies also have IgG and IgM capable of binding SLA. Inactivating SLA to eliminate their contribution to human anti-pig immunity is problematic because the SLA are key to helping protect the organ from infection and cancer. The objectives of this grant are to: (i) improve our understanding of the frequency with which patients have anti-SLA antibodies; (ii) determine the origin of B cells which produce those antibodies, and (iii) define their susceptibility to a B-cell depleting therapy (Rituximab). Our approach is innovative because it will develop novel tools that build on similar approaches that have been successful in the setting of allotransplantation. Our rationale is that this knowledge will bring us bring us closer to the use of pigs as an organ source by helping to better match donors with recipients. In Specific Aim 1 we will create recombinant SLA, produce a bead array of these proteins, and use the array to screen 500 human sera for the presence of SLA antibodies. The focus of Specific Aim 2.1 is to screen 25 patients before and after Rituximab treatment with the SLA bead array to determine if those xenoreactive antibodies diminish. Specific Aim 2.2 will identify and characterize SLA-specific B cells to determine if natural antibodies account for some of the SLA reactivity. This technology development will contribute significantly to the ability to match pig donors with human recipients by avoiding or eliminating SLA-specific antibodies. This project will rely on several components of this award to achieve its goals. Core C will provide bead arrays for serum screening and SLA tetramers with which to analyze B cells provided by Core B. Project 2 investigators and Core D will help with the sorting and analyses of tetramer-stained B cells. Project 1 will provide Rituximab samples and data and assist in comparisons to their work to determine if B cell depletion affects SLA-specific antibodies and naturally occurring anti-glycan antibodies similarly.

项目 3：开发识别异种移植反应性 B 细胞的新型试剂项目概要可用供体器官的短缺是当今器官移植面临的最严峻的挑战。猪是被认为是替代组织的有前途的来源。不幸的是，异种移植，共享跨物种的器官，尚未应用于临床，因为人类会产生强烈的体液免疫反应朝向猪组织。我们的长期目标是通过预防人类抗体的结合。该项目的中心假设是猪的主要组织相容性复合体 (MHC) 蛋白，在猪体内被称为猪白细胞抗原 (SLA)，有助于异种抗原性。我们已经证明，具有人类白细胞抗原 (HLA) 抗体的患者经常会发生交叉反应与同源 SLA。我们还发现，大约 25% 的人群缺乏 HLA 抗体还具有能够结合 SLA 的 IgG 和 IgM。停用 SLA 以消除其对人类抗猪免疫力存在问题，因为 SLA 是帮助保护器官免受感染的关键和癌症。这笔赠款的目的是：（i）提高我们对频率的了解患者有抗SLA抗体； (ii) 确定产生这些抗体的 B 细胞的来源，以及 (iii) 确定他们对 B 细胞耗竭疗法（利妥昔单抗）的敏感性。我们的方法是创新的，因为它将开发新的工具，这些工具建立在类似方法的基础上，这些方法在以下领域取得了成功同种异体移植。我们的理由是，这些知识将使我们更接近利用猪作为动物通过帮助更好地匹配捐赠者和接受者来获取器官来源。在具体目标 1 中，我们将创建重组 SLA，产生这些蛋白质的珠阵列，并使用该阵列筛选 500 份人类血清中是否存在 SLA 抗体。具体目标2.1的重点是筛查利妥昔单抗治疗前后的25名患者使用 SLA 珠阵列来确定这些异种反应抗体是否减少。具体目标 2.2 将确定并对 SLA 特异性 B 细胞进行表征，以确定天然抗体是否能解释某些 SLA 反应性。这项技术的发展将大大有助于将猪捐赠者与人类捐赠者相匹配的能力通过避免或消除 SLA 特异性抗体来治疗接受者。该项目将依赖于以下几个组件该奖项旨在实现其目标。 Core C 将提供用于血清筛查的微珠阵列和 SLA 四聚体用于分析 Core B 提供的 B 细胞。项目 2 的研究人员和 Core D 将帮助进行分类和四聚体染色的 B 细胞分析。项目1将提供利妥昔单抗样品和数据并协助与他们的工作进行比较，以确定 B 细胞耗竭是否会影响 SLA 特异性抗体，并且自然地类似地出现抗聚糖抗体。