Molecular and functional investigation of the role of HLA-F in immune regulation

HLA-F在免疫调节中作用的分子和功能研究

• 批准号: 10503676
• 负责人: Erin June Adams
• 金额: $ 68.3万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-06 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10503676
• 关键词: 3-Dimensional; Address; Adenoviruses; Adopted; Amino Acids; Antibodies; Antigens; Applications Grants; Area; Autoimmune Diseases of the Nervous System; Autoimmunity; Binding; Biological Assay; Biophysics; Cell surface; Cells; Clone Cells; Data; Decidua; Disease; Effector Cell; Environment; Family; Family member; Funding; Genetic Polymorphism; Goals; HIV; HIV Infections; Health; Homeostasis; Human; Immune; Immunity; Infection; Invaded; Investigation; KLRD1 gene; Length; Ligands; Link; Major Histocompatibility Complex; Malignant Neoplasms; Methods; Modeling; Molecular; Molecular Chaperones; Molecular Conformation; Natural Killer Cells; Peptides; Play; Post-Translational Protein Processing; Pregnancy; Production; Protein Isoforms; Proteins; Publishing; RNA Splicing; Reagent; Receptor Cell; Recombinants; Reproduction; Role; Source; Specificity; Structure; T-Lymphocyte; Testing; Variant; Viral; Work; biophysical analysis; cancer cell; cell type; conformer; fetal; immunoregulation; member; pathogen; protein complex; receptor; three dimensional structure; trafficking; tumor

Project Summary/Abstract HLA-F is a nonclassical class I MHC (Ib) molecule that has been found expressed on a variety of cancers, shown to play a role in HIV and adenoviral infection, the neurological autoimmune disease ALS and is expressed throughout pregnancy. Despite the potential importance of this protein in these conditions, little is known about this molecule in terms of its function or even in which conformational state it is expressed. We have recently shown that, in addition to being expressed as a heavy chain only state, or open conformer (HLA-FOC), HLA-F can also be expressed as a bon fide peptide presenting molecule, associated with the β2m subunit (pHLA-F). Peptides are presented in an unconventional way, with the N-terminus not anchored within the groove and the potential for post-translational modifications featuring in peptide anchoring. Despite these advances, there remains much unknown about how these conformer states are regulated, how it engages its various receptors in each of these conformer states, and the role of HLA-F in its various environments of tumor surveillance, autoimmunity and reproduction. Thus, the aims of this proposal focus on addressing these questions and are: Aim 1: To investigate, structurally and functionally, the various conformer states that HLA-F adopts in human health and disease. We will pursue structural studies of the HLA-F isoforms to understand how these two states differ from each other. Using conformer-specific antibodies, we will determine what cell types express which (or both) forms and how this differs between healthy and disease cells. We will also pursue peptide elution studies from a range of human sources to determine if the peptide repertoire shifts depending on cellular origin or disease. Aim 2: To identify and analyze the factors that regulate the production or interchange of HLA- F conformers and splice forms in a cell. We will explore the cellular factors that may play a role in switching HLA-F between peptide-loaded and HLA-FOC as well as an intriguing splice variant of HLA-F of unknown function. Finally, in Aim 3 we seek to establish the receptor repertoire that engage HLA-F in its various conformer states, determine the molecular basis for their association and study the functional consequences of their binding. We will employ the structural, biophysical and functional expertise of the Adams lab to determine the receptor repertoire that engage these conformer states of HLA-F and study them at the functional and molecular level.

项目摘要/摘要 HLA-F是一种非经典的I MHC（IB）分子，已在各种癌症上表达， 证明在HIV和腺病毒感染中起作用，神经自身免疫性疾病ALS并表达 整个怀孕。尽管该蛋白在这些条件下可能具有重要意义，但对 该分子就其功能甚至表达的构象状态而言。我们最近有 表明，除了表达为重链状态或开放式构象异构体（HLA-FOC）外，HLA-F之外 也可以表示为与β2M亚基（PHLA-F）相关的典型肽呈现分子。 肽以非常规的方式呈现，N末端没有锚定在凹槽内，并且没有锚固 在辣椒锚定中进行翻译后修饰的潜力。尽管有这些进步， 对于这些构象异构态的调节，如何与各种接收器保持在一起，仍然不知道 在每个构象异构态中，以及HLA-F在其各种肿瘤监测环境中的作用， 自身免疫性和繁殖。这是该提案的目的，重点是解决这些问题，是： 目的1：在结构和功能上调查HLA-F所采用的各种构象异构体 人类健康和疾病。我们将进行HLA-F同工型的结构研究，以了解这些方法 两个状态彼此不同。使用构象异构抗体，我们将确定哪种细胞类型表达 哪种形式以及健康细胞和疾病细胞之间的差异。我们还将纯化肽洗脱 来自各种人类来源的研究，以确定胡椒曲目是否根据细胞起源而移动 或疾病。目标2：识别和分析调节HLA-生产或互换的因素 F中的F构象和剪接形成。我们将探索可能在切换中起作用的细胞因素 肽负载和HLA-FOC之间的HLA-F以及未知功能的HLA-F的有趣剪接变体。 最后，在目标3中，我们试图建立吸引HLA-F的受体曲目 状态，确定其关联的分子基础，并研究 他们的约束。我们将采用Adams实验室的结构，生物物理和功能专业知识来确定 接合HLA-F的这些符合符号并在功能上研究它们的受体曲目 分子水平。