Molecular and Structural Studies of Antibody Diversity Mechanisms

抗体多样性机制的分子和结构研究

基本信息

批准号：
9982985
负责人：
Vaughn Vasil Smider
金额：
$ 39.94万
依托单位：
APPLIED BIOMEDICAL SCIENCE INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-30 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9982985
关键词：
Active Sites Amino Acids Antibodies Antibody Diversity Antibody Formation Antibody Repertoire Antigen Targeting Antigens Area Binding Biochemical Biological Biological Response Modifiers Cattle Cells Cellular Assay Color Complementarity Determining Regions Complex Cysteine Data Diagnostic Diagnostics Research Disulfides Electron Microscopy Enzyme Activation Enzymes Epitopes Event Fab Immunoglobulins Family Flow Cytometry Gene Mutation Genes Genetic Genetic Structures HIV HIV Antibodies HIV-1 Human Immune response Immunization Immunize Immunoglobulin G Immunoglobulin Somatic Hypermutation In Vitro Infectious Agent Investigational Therapies Ion Channel Label Left Length Light Maps Matrix Metalloproteinases Mediating Medicine Metalloproteases Methods Modeling Molecular Molecular Genetics Molecular Structure Mus Mutagenesis Mutate Mutation Pathway interactions Pattern Play Polysaccharides Process Property Proteins Reagent Recombinants Role Site-Directed Mutagenesis Structure Surface System Tertiary Protein Structure Testing Therapeutic Time V(D)J Recombination Vaccine Design Vaccines Vertebrates Viral Virus activation-induced cytidine deaminase antibody engineering antigen binding antigen challenge base combinatorial disulfide bond electron crystallography in vivo insight knob protein neutralizing antibody novel response therapeutic candidate vaccine response

项目摘要

Abstract Antibodies are extremely important as the primary mediators of the immune response to infectious agents and vaccines, but also as recombinant molecules used as therapeutics, diagnostics, and research reagents. Thus, understanding the mechanisms by which antibodies form, bind, and neutralize their targets is very important in multiple biomedical areas. Most vertebrate antibody repertoires form their diversity through V(D)J recombination, where combinatorial rearrangement of V, D, and J genes form a vast repertoire where the complementarity determining regions (CDRs) form a relatively flat binding surface for interaction with antigen. Remarkably, cows appear to have a different mechanism for generating diversity and binding antigen; cow antibodies have particularly long CDR H3 regions, a subset of which can be over 70 amino acids long, which form -ribbon “stalk” and disulfide-bonded “knob” minidomains that protrude far from the typical antibody surface. Remarkably, antibodies from this repertoire can potently and broadly neutralize HIV, whereas normal human or mouse antibody repertoires cannot. Therefore, they have unique abilities to bind and neutralize particularly challenging antigens. Here we propose studies to understand in molecular detail the genetic mechanisms underlying formation of these antibodies, the binding properties of the stalk and knob minidomains, and the unique potential of these antibodes to bind bivalently and bispecifically. These studies will provide insight into mechanisms of viral neutralization generally, and particularly HIV neutralization. Additionally, given the potential of these antibodies to bind recessed epitopes, we will generate additional antibodies against particularly challenging antigens like enzymatic active sites and ion channels. We will employ mutagenesis, binding, and structural methods, as well as cellular assays to evaluate the unique properties of these antibodies. Understanding this novel class of antibodies in detail could provide fundamental insights needed for effective vaccine design as well as discovery and engineering of antibodies against some of the most challenging targets in biomedicine. Similarly, our unique antibodies discovered and characterized in this proposal could eventually become therapeutic candidates themselves.

抽象的抗体非常重要，因为免疫响应的主要介体传染性剂和疫苗，也是用作治疗的重组分子，诊断和研究试剂。这是了解抗体的机制在多个生物医学领域，形式，结合和中和目标非常重要。最多脊椎动物抗体曲目通过V（d）J重组形成其多样性，其中 V，D和J基因的组合重排构成了一个广阔的曲目互补性确定区域（CDR）形成相对平坦的结合表面以进行相互作用与抗原。值得注意的是，牛似乎具有产生多样性的不同机制和结合抗原；牛抗体具有特别长的CDR H3区域，其中一个子集可以长70多个氨基酸，形成-丝带“茎”，并二硫键键入“旋钮” 远离典型抗体表面的小型膜。值得注意的是，抗体来自该曲目可能会潜在地和广泛中和HIV，而正常的人或小鼠抗体库不能。因此，它们具有绑定和中和的独特能力特别具有挑战性的抗原。在这里，我们建议研究以分子详细了解这些抗体的遗传机制，茎的结合特性和旋钮的初学酶，以及这些抗体以二人结合的独特潜力和双方。这些研究将洞悉病毒神经化的机制通常，尤其是HIV神经化。此外，考虑到这些潜力结合嵌入式表位的抗体，我们将特别生成针对的其他抗体具有挑战性的抗原，例如酶活性位点和离子通道。我们将采用诱变，结合方法和结构方法以及细胞评估，以评估这些抗体。了解这一新颖的抗体可以详细提供有效疫苗设计以及发现和工程所需的基本见解针对生物医学中一些最具挑战性靶标的抗体。同样，我们的独特之处在该提案中发现和表征的抗体最终可能成为治疗候选人自己。