Small Animals Core

小动物核心

• 批准号: 10365961
• 负责人: S. Munir ALAM
• 金额: $ 29.39万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-09 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10365961
• 关键词: Address; Affinity; Alleles; Animal Model; Animals; Antibodies; Antibody Repertoire; Antigens; B cell repertoire; B-Lymphocytes; Biological Assay; Boston; Bypass; Cell Compartmentation; Cell Maturation; Complex; Development; Distal; Epitopes; Evaluation; Evolution; Exclusion; Exons; Genes; Genetic Recombination; HIV; HIV-1; Human; Immunization; Immunize; Immunoglobulin Genes; Immunoglobulins; In Vitro; Individual; Knock-in; Lead; Light; Mature B-Lymphocyte; Membrane; Methods; Modeling; Mus; Nucleotides; Pediatric Hospitals; Physiological; Population; Research; Research Personnel; Scheme; Series; Source; Specificity; System; Testing; V(D)J Recombination; central tolerance; design; efficacy testing; env Gene Products; experience; experimental study; falls; immunogenic; in vivo; mouse model; neutralizing antibody; programs; screening

Abstract - Small Animal Models Core The objective of the Small Animals Model Core is to generate a series of mouse models that express the unmutated common ancestor (UCA) or intermediate antibodies (IAs) of the DH511 lineage of broadly neutralizing antibody (bnAb). These mouse models will be used to test the efficacy of immunogens that are designed to elicit the DH511 lineage of bnAbs. Toward this end, the Animal Model Core will generate two types of mouse models. The first type of model expresses pre-rearranged V(D)J exons of the DH511 antibodies. Owing to allelic exclusion, the pre-rearranged DH511 V(D)J exon will inhibit the rearrangement of endogenous mouse immunoglobulin (Ig) loci. As a result, B cells in these mice will express predominantly DH511 antibodies. One common problem for this type of mouse model is that tolerance control mechanisms delete B cells expressing the knock-in human Ig genes. To overcome this hurdle, we have developed a method to express human antibodies conditionally in mature B cells, thereby circumventing tolerance control during B cell maturation. If necessary, we will employ this conditional expression system to generate DH511 mouse models. Since this type of model provides a large population of B cells expressing DH511UCA or IAs, it would serve as a sensitive assay for the initial evaluation of immunogens. However, the system does not recapitulate the complexity of physiological B cell repertoire in at least two major respects. First, the unique UCA expressed in the mouse model may not be present in a fraction of human populations. Second, the system lacks competing antibodies for irrelevant epitopes. To address these limitations, the Animal Model Core will generate a second type of mouse model where the VH3-15, D3-3 and JH6 gene segments of DH511 undergo de novo V(D)J recombination. Due to junctional diversity, the recombination will create a wide range of CDR H3s, some of which may be suitable for the development of DH511-like antibodies. Moreover, the system does not preclude the rearrangements of endogenous mouse Ig gene segments. Therefore, the B cell repertoire of this mouse model will consist of potential DH511 precursors as well as other human VH3-15/D3-3/JH6 and mouse antibodies. Immunization of this mouse model could assess the ability of the immunogen to select for and mature DH511 precursors in the context of complex antibody repertoires.

摘要 - 小动物模型核心 小动物模型核心的目的是生成一系列表达的小鼠模型 DH511谱系的未分离的共同祖先（UCA）或中间抗体（IAS） 中和抗体（BNAB）。这些鼠标模型将用于测试免疫原子的功效 旨在引起BNAB的DH511谱系。为此，动物模型核心将产生两种类型 鼠标模型。第一种模型表达了DH511抗体的预先键入V（d）J外显子。 由于等位基因排除，预先矫正的DH511 V（d）J外显子将抑制内源性的重排 小鼠免疫球蛋白（IG）基因座。结果，这些小鼠中的B细胞主要表达DH511 抗体。这种类型的鼠标模型的一个常见问题是，公差控制机制删除了b 表达敲入人Ig基因的细胞。为了克服这一障碍，我们开发了一种方法 在成熟的B细胞中有条件地表达人类抗体，从而规避B细胞期间的耐受性控制 成熟。如有必要，我们将采用此条件表达系统生成DH511鼠标模型。 由于这种类型的模型提供了表达DH511UCA或IAS的大量B细胞，因此它将作为 用于初始评估免疫原子的敏感测定法。但是，该系统不会概括 在至少两个主要方面，生理B细胞库的复杂性。首先，独特的UCA在 小鼠模型可能不存在于人类人群的一部分中。其次，系统缺乏竞争 无关表位的抗体。为了解决这些局限性，动物模型核心将产生第二个 小鼠模型的类型，其中DH511的VH3-15，D3-3和JH6基因段进行了DE NOVO V（D）J 重组。由于连接性的多样性，重组将产生各种CDR H3S，其中一些 可能适合开发DH511样抗体。而且，系统不排除 内源性小鼠Ig基因段的重排。因此，该小鼠的B细胞曲目 模型将由潜在的DH511前体以及其他人VH3-15/D3-3/JH6和小鼠组成 抗体。该小鼠模型的免疫可以评估免疫原选择的能力， 成熟的DH511前体在复杂的抗体库中。