Tracking Immune Activation by Stress Proteins In Vivo

体内应激蛋白追踪免疫激活

• 批准号: 7341718
• 负责人: CHRISTOPHER A PENNELL
• 金额: $ 25.25万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7341718
• 关键词: Accounting; Address; Adjuvant; Adjuvanticity; Adoptive Transfer; Affect; Amino Acids; Animal Model; Antigens; Binding; Biology; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell Communication; Cell Surface Receptors; Cell Survival; Cells; Chickens; Chimeric Proteins; Data; Dendritic Cells; Dopachrome isomerase; Employee Strikes; Event; Flow Cytometry; Fluorochrome; Frequencies; Generations; Goals; Heat shock proteins; Hour; Immune; Immune response; Immune system; Immunization; Kinetics; Life; Link; Longevity; Maintenance; Measures; Mediating; Memory; Monoclonal Antibodies; Mus; Mycobacterium tuberculosis; Numbers; Peptides; Phenotype; Production; Proliferating; Proteins; Reporting; Research Personnel; Signal Transduction; Stress-Induced Protein; System; T memory cell; T-Cell Activation; T-Lymphocyte; Testing; Time; Transgenic Organisms; Tumor Immunity; Vaccines; Viral; base; cancer therapy; cell type; cytokine; day; density; in vivo; insight; lymph nodes; melanocyte; novel; programs; receptor; research study; response; stress protein; uptake; vaccine development

Vaccines containing peptides linked to stress proteins are the most potent known T cell immunogens. However, the precise mechanisms by which they activate immune cells remain unclear. To address this, we will track activated dendritic cells (DCs) and CD8+ T cells in mice immunized with antigens fused to the stress protein hsp70, and determine which cytokines and cell surface receptors are required for their responses. Our preliminary studies reveal hspTO fusion vaccines elicit much more potent and durable specific CD8+ T cell responses than antigen alone or with other adjuvants. Our underlying hypothesis is that antigen/hsp70 fusions are potent vaccines because they stimulate DCs to provide an optimal microenvironment for CD8+ T cell activation. This microenvironment is defined by high antigen density and strong costimulation. To test this, we will track in vivo DC and CD8+ T cell responses in draining lymph nodes, and the resultant effector and memory CD8+ T lymphocytes in the periphery. Novel aspects of our proposal include: 1) quantifying the amount of antigen presented by activated DCs, 2) the effect hsp70 fusion proteins have on DC lifespan, 3) determining which of the several reported hsp70-receptors are necessary or sufficient for delivering activation signals to DCs in vivo, and 4) determining the costimulatory and cytokines required for the profound effect our vaccine has CD8+ T cell memory. The results of these experiments will allow us to manipulate stress protein-induced responses rationally with the ultimate goal of maximizing their use for cancer therapy.

含有与应激蛋白连接的肽的疫苗是已知最有效的T细胞免疫原。 然而，它们激活免疫细胞的确切机制仍不清楚。为了解决这个问题，我们 将追踪用与应激融合的抗原免疫的小鼠中活化的树突状细胞（DC）和CD 8 + T细胞 蛋白HSP 70，并确定其反应所需的细胞因子和细胞表面受体。 我们的初步研究表明，hspTO融合疫苗可诱导更有效和持久的特异性CD 8 + T细胞 与单独的抗原或与其它佐剂一起的细胞应答相比。我们的基本假设是抗原/hsp 70 融合物是有效的疫苗，因为它们刺激DC为CD 8 + T细胞提供最佳的微环境。 细胞激活这种微环境由高抗原密度和强共刺激定义。 为了验证这一点，我们将跟踪引流淋巴结中的体内DC和CD 8 + T细胞应答，并将结果与淋巴结中的DC和CD 8 + T细胞应答进行比较。 外周中的效应和记忆CD 8 + T淋巴细胞。我们建议的新方面包括：1） 定量由活化的DC呈递的抗原的量，2）hsp 70融合蛋白对 DC寿命，3）确定几种报道的hsp 70受体中的哪一种对于DC寿命是必需的或足够的。 在体内将活化信号递送至DC，以及4）确定活化所需的共刺激和细胞因子。 我们的疫苗具有CD 8 + T细胞记忆的深远影响。 这些实验的结果将使我们能够合理地操纵应激蛋白诱导的反应， 最终目标是最大限度地将其用于癌症治疗。