Anti-GD3 NKT cells as effector cells against melanoma

抗 GD3 NKT 细胞作为抗黑色素瘤的效应细胞

• 批准号: 6752082
• 负责人: PAUL B CHAPMAN
• 金额: $ 28.01万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6752082
• 关键词: CD1 molecule; Golgi apparatus; T cell receptor; T lymphocyte; antigen antibody reaction; antigen presentation; ceramides; confocal scanning microscopy; cytokine; gangliosides; laboratory mouse; melanoma; natural killer cells; neoplasm /cancer immunology; receptor mediated endocytosis; tissue /cell culture; tumor antigens; vesicle /vacuole

DESCRIPTION (provided by applicant): GD3 ganglioside is expressed on neuroectodermal tissue and on tumors such as melanoma, sarcoma, and small-cell lung cancer. Antibodies against GD3 can shrink melanoma in rodents and patients. In clinical trials designed to immunize patients against GD3 in the adjuvant setting, we saw surprisingly low relapse rates that did not correlate with induction of anti-GD3 antibodies. Since GD3 is not presented by MHC class I or class II, we considered the alternative possibility that immunization might induce a NKT response against GD3. We have induced CD1-restricted NKT cells against GD3 in mice and in this project; we propose to characterize further the anti-GD3 murine NKT cell response. Specific Aim 1 - Define the T cell receptor usage, cytokine profile, and specificity of mouse CD1-restricted NKT cells against GD3. Most murine CD1-restricted NKT cells described to date recognize glycolipids derived from bacteria or invertebrates. Much less is known about CD1-restricted NKT cells that recognize self-glycolipids expressed on mammalian cells. Specific Aim 2 - Characterize how GD3 is loaded on to mouse CD1. In the human system, some CD1-restricted glycolipid antigens require internalization and loading in acidified late endosomes; other CD1-restricted glycolipids are loaded in post-Golgi vesicles not requiring acidification. Little is known about how self gangliosides are loaded on to CD1 although in the human system there is some evidence that GM1 ganglioside does not even require internalization to be presented. Using the mouse system, we will characterize how GD3 is loaded on to mouse CD1 assessing requirements for internalization, acidification of endosomes and Golgi function. We plan to map the intracellular processing pathway using confocal immunomicroscopy. Specific Aim 3 -Test the hypothesis that mouse NKT cells against GD3 can mediate anti-tumor effects in an antigen-specific manner. CD1-restricted NKT cells activated by alpha-galactosylceramide (from marine invertebrates) can mediate antitumor effects in a manner that does not appear to be antigen-specific. It is not known if CD1-restricted NKT cells specific for an antigen expressed on tumor cells can have anti-tumor effects. We will test the ability of CD1-restricted NKT cells against GD3 to lyse GD3+ mouse melanoma in vitro and to reject GD3+ mouse tumors in vivo. This will provide initial support for an immunotherapeutic approach based on antigen-specific NKT cells.

描述（由申请人提供）：GD3神经节苷脂在神经外胚层组织和肿瘤（例如黑色素瘤、肉瘤和小细胞肺癌）上表达。 GD3 抗体可以缩小啮齿动物和患者的黑色素瘤。在设计用于在辅助环境中对患者进行 GD3 免疫的临床试验中，我们发现复发率令人惊讶地低，这与抗 GD3 抗体的诱导无关。由于 GD3 不由 I 类或 II 类 MHC 呈递，我们考虑了免疫可能诱导针对 GD3 的 NKT 反应的另一种可能性。在本项目中，我们在小鼠体内诱导了针对 GD3 的 CD1 限制性 NKT 细胞；我们建议进一步表征抗 GD3 小鼠 NKT 细胞反应。具体目标 1 - 定义 T 细胞受体的使用、细胞因子谱以及小鼠 CD1 限制性 NKT 细胞针对 GD3 的特异性。迄今为止，大多数鼠类 CD1 限制性 NKT 细胞都能识别源自细菌或无脊椎动物的糖脂。对于识别哺乳动物细胞上表达的自身糖脂的 CD1 限制性 NKT 细胞知之甚少。具体目标 2 - 描述如何将 GD3 加载到鼠标 CD1 上。在人体系统中，一些 CD1 限制性糖脂抗原需要内化并装载到酸化的晚期内体中；其他 CD1 限制性糖脂装载在不需要酸化的高尔基体后囊泡中。尽管在人体系统中，有一些证据表明 GM1 神经节苷脂甚至不需要内化才能呈现，但关于自身神经节苷脂如何加载到 CD1 上的情况知之甚少。使用小鼠系统，我们将描述 GD3 如何加载到小鼠 CD1 上，评估内化、内体酸化和高尔基体功能的要求。我们计划使用共聚焦免疫显微镜绘制细胞内加工途径。具体目标 3 - 测试针对 GD3 的小鼠 NKT 细胞可以以抗原特异性方式介导抗肿瘤作用的假设。由 α-半乳糖神经酰胺（来自海洋无脊椎动物）激活的 CD1 限制性 NKT 细胞可以以似乎不具有抗原特异性的方式介导抗肿瘤作用。目前尚不清楚针对肿瘤细胞上表达的抗原具有特异性的 CD1 限制性 NKT 细胞是否具有抗肿瘤作用。我们将测试针对 GD3 的 CD1 限制性 NKT 细胞在体外裂解 GD3+ 小鼠黑色素瘤和在体内排斥 GD3+ 小鼠肿瘤的能力。这将为基于抗原特异性 NKT 细胞的免疫治疗方法提供初步支持。