Charaterization of the Expression and Ligands of KIR3DS1

KIR3DS1 表达和配体的表征

• 批准号: 7965595
• 负责人: Daniel W. McVicar
• 金额: $ 12.92万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7965595
• 关键词: Acquired Immunodeficiency Syndrome; Affect; Alleles; Antibodies; Antiviral Response; Binding; Biochemical; Cell Line; Cells; Cytoplasmic Tail; Disease; Dissection; Epitopes; Extracellular Domain; Genes; Goals; HIV; HIV-1; HLA-Bw4; Immune response; Immune system; Individual; Infection; KIR3DS1; Lentivirus Vector; Ligand Binding; Ligands; NK Cell Activation; NK cell receptor NKB1; Natural Killer Cells; Play; Proteins; Regulation; Reporter; Role; Screening procedure; Signal Transduction; Stress; Structure; System; T-Cell Receptor; killer immunoglobulin-like receptor; receptor; therapeutic development

KIR3DS1 is presumed to be an NK cell activation receptor. The gene has many (>40) inhibitory alleles, known as KIR3DL1. Various KIR3DL1 subtypes have been shown to interact directly with HLA-I proteins with the Bw4 public epitope. Regardless of its similarity to KIR3DL1, and its established role in HIV disease, KIR3DS1 has only recently been shown to be expressed on NK cells and no ligand has been described. Toward an understanding of KIR3DS1 ligand binding we developed a reporter system for the engagement of KIR3DS1. The system uses KIR3DS1 fused to the T cell receptor zeta signaling chain. In addition, we have established HLA Bw4 (B5701) expressing cell lines. These cell lines are infected with lentiviral vectors and combined with the reporter line. Thus far, lentiviral infection does not result in the activation of the KIR3DS1 reporter system. In addition, we have induced stress in these Bw4 expressing cells before combining them with the reporter cells. We also have identified key residues of KIR3DS1 that do not exist in KIR3DL1. Through mutagensis we have shown that W138 of KIR3DS1 is critically involved in the lack of recognition of HLA-Bw4 by KIR3DS1. Reversion of W138 to G found in KIR3DL1 confers Bw4 binding without affecting the expression or antibody recognition of the receptor.

Kir3ds1被认为是NK细胞激活受体。该基因有许多（> 40）抑制性等位基因，称为KIR3DL1。已显示各种KIR3DL1亚型与HLA-I蛋白与BW4公共表位直接相互作用。无论它与Kir3DL1的相似性及其在HIV疾病中的确定作用，Kir3DS1最近才显示在NK细胞上表达，并且没有描述配体。为了了解Kir3DS1配体结合，我们开发了一个用于参与Kir3DS1的记者系统。该系统使用与T细胞受体Zeta信号链融合的KiR3DS1。此外，我们已经建立了表达细胞系的HLA BW4（B5701）。这些细胞系被慢病毒载体感染，并与记者线结合。到目前为止，慢病毒感染并未导致KIR3DS1报告基因系统的激活。此外，在将这些BW4表达细胞与报告细胞结合之前，我们诱导了应力。我们还确定了KIR3DS1的关键残基，这些残基在KiR3DL1中不存在。通过诱变，我们已经表明，Kir3ds1的W138与Kir3ds1对HLA-BW4的识别缺乏认识。在KiR3DL1中发现的W138的恢复为G，赋予BW4结合而不影响受体的表达或抗体识别。