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

• 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
• 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

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：开发新试剂以鉴定异种移植反应性B细胞 项目摘要 缺乏可用的供体器官是当今器官移植面临的最关键的挑战。猪是 被认为是有希望的替代组织的来源。不幸的是，异种移植，共享 跨物种的器官在临床上没有应用，因为人类发挥强烈的体液免疫反应 朝向猪组织。我们的长期目标是通过防止 人类抗体的结合。该项目的中心假设是猪主要的组织相容性 复合物（MHC）蛋白，称为猪中的猪白细胞抗原（SLA），有助于异种抗原性。 我们已经表明，患有针对人类白细胞抗原（HLA）抗体的患者经常交叉反应 与同源SLA。我们还发现，缺乏HLA的人口中约有25％ 抗体还具有能够结合SLA的IgG和IgM。使SLA失活以消除其对 人类抗铅免疫是有问题的，因为SLA是帮助保护器官免受感染的关键 和癌症。这笔赠款的目标是：（i）提高我们对频率的理解 患者具有抗SLA抗体； （ii）确定产生这些抗体的B细胞的起源，以及（iii） 定义它们对B细胞耗尽疗法（利妥昔单抗）的敏感性。我们的方法是创新的，因为它将 开发基于在类似方法的基础上成功的新颖工具 同种异体移植。我们的理由是，这些知识将使我们更接近使用猪作为 器官来源通过帮助更好地匹配接收者来更好地匹配捐助者。在特定目标1中，我们将创建重组 SLA，产生这些蛋白质的珠阵列，并使用阵列筛选500人血清的存在 SLA抗体。特定目标2.1的重点是在利妥昔单抗治疗前后筛查25名患者 使用SLA珠阵列来确定那些异种反应性抗体是否减小。特定目标2.2将确定 并表征SLA特异性B细胞，以确定天然抗体是否解释了某些SLA反应性。 这项技术开发将极大地促进与人类匹配猪捐赠者的能力 通过避免或消除SLA特异性抗体，接受者。该项目将依靠几个组成部分 这个奖项是为了实现其目标。 Core C将提供血清筛查的珠阵列，用于血清筛查和SLA四聚体 要分析由Core B提供的B细胞。项目2研究者和核心D将有助于分类和 四聚体染色的B细胞的分析。项目1将提供利妥昔单抗样本和数据，并协助 与他们的工作进行比较，以确定B细胞耗竭是否影响SLA特异性抗体和自然 发生抗甘油抗体类似。