Molecular Identification of an Endogenous Adjuvant

内源性佐剂的分子鉴定

• 批准号: 7027763
• 负责人: YAN SHI
• 金额: $ 15.82万
• 依托单位: UNIVERSITY OF CALGARY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2007-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7027763
• 关键词: cell communication molecule; cell death; cytotoxic T lymphocyte; dendritic cells; immune response; immunologic assay /test; immunomodulators; laboratory mouse; leukocyte activation /transformation; physical separation; small interfering RNA; urate

DESCRIPTION (provided by applicant): Cell injury leads to the induction of immune responses under a variety of pathological circumstances. It has been proposed that cell death provides danger signals that help simulate immunity against cellular antigens. With the recent molecular identification of uric acid as one of the participating factors, the identity of another immune regulator present in the cytosol remains elusive. The specific hypothesis of the project is that cellular disintegration during cell injury releases molecules that are normally sequestered under physiological conditions, and that once released, these substances exert adjuvant activities by activating antigen presenting cells. We base our hypothesis on the following observations 1.) The cytosol from injured cells has been shown to have adjuvant activity in CTL induction; 2.) A high molecular weight substance(s) (40 to 100 KD) purified from sizing columns activates antigen presenting cells. The goal of this study is to biochemically identify and characterize this high molecular weight substance. We have two specific aims. 1. Molecular identification of the high molecular weight substance(s). We will use the combined approach of chromatography/mass spectrometry for identification. We will monitor adjuvant activities in HPLC fractions by CTL induction/dendritic cell activation. Substantially purified active fractions will be analyzed by mass spectrometers. 2. Characterization of high molecular weight adjuvant activities. This part of the project will focus on the characterization of the substance identified in Aim 1. It will be either purified directly from cells to homogeneity or expressed/synthesized in appropriate cells. The substance will be tested for its ability to promote CTL responses, dendritic cell activation and other immuno-enhancing capacities. We will also attempt to make antibodies against the substance or use siRNA interference to block the adjuvant activities. In all assays, the substance will be tested side by side with uric acid crystals for a direct comparison of their potencies.

描述（由申请人提供）：细胞损伤导致在多种病理情况下诱导免疫反应。有人提出，细胞死亡提供了有助于模拟针对细胞抗原的免疫力的危险信号。随着最近分子鉴定尿酸作为参与因素之一，细胞质中存在的另一种免疫调节剂的身份仍然难以捉摸。该项目的具体假设是，细胞损伤期间的细胞崩解释放出通常在生理条件下隔离的分子，一旦释放，这些物质通过激活抗原呈递细胞发挥佐剂活性。我们的假设基于以下观察：1.) 来自受损细胞的胞质溶胶已被证明在 CTL 诱导中具有佐剂活性； 2.) 从分级柱纯化的高分子量物质（40 至 100 KD）可激活抗原呈递细胞。本研究的目的是通过生化方法鉴定和表征这种高分子量物质。我们有两个具体目标。 1. 高分子量物质的分子鉴定。我们将使用色谱/质谱联用方法进行鉴定。我们将通过 CTL 诱导/树突状细胞激活来监测 HPLC 级分中的佐剂活性。将通过质谱仪分析基本上纯化的活性级分。 2.高分子量佐剂活性的表征。该项目的这一部分将重点关注目标 1 中确定的物质的表征。它将直接从细胞中纯化至均质，或在适当的细胞中表达/合成。该物质将测试其促进 CTL 反应、树突状细胞激活和其他免疫增强能力的能力。我们还将尝试制造针对该物质的抗体或使用 siRNA 干扰来阻断佐剂活性。在所有测定中，该物质将与尿酸晶体一起进行测试，以直接比较它们的效力。

项目成果

A role of IgM antibodies in monosodium urate crystal formation and associated adjuvanticity.

IgM 抗体在尿酸钠晶体形成和相关佐剂作用中的作用。

• 发表时间: 2009-02-15
• 作者: Kanevets, Uliana;Sharma, Karan;Dresser, Karen;Shi, Yan
• 通讯作者: Shi, Yan

Receptor-independent, direct membrane binding leads to cell-surface lipid sorting and Syk kinase activation in dendritic cells.

不依赖于受体的直接膜结合导致树突状细胞中的细胞表面脂质分选和 Syk 激酶激活。

• DOI: 10.1016/j.immuni.2008.09.013
• 发表时间: 2008-11-14
• 期刊: Immunity
• 影响因子: 32.4
• 作者: Ng, Gilbert;Sharma, Karan;Ward, Sandra M.;Desrosiers, Melanie D.;Stephens, Leslie A.;Schoel, W. Michael;Li, Tonglei;Lowell, Clifford A.;Ling, Chang-Chun;Amrein, Matthias W.;Shi, Yan
• 通讯作者: Shi, Yan