Enhancing Immunotherapeutic Value of Lymphocytes by Controlling Apoptosis

通过控制细胞凋亡增强淋巴细胞的免疫治疗价值

• 批准号: 7592316
• 负责人: John E Coligan
• 金额: $ 57.83万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 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/7592316
• 关键词: Apoptosis; Apoptotic; Biological Assay; CD95 Antigens; Cell Death; Cell Line; Cells; Cessation of life; Class; Cultured Cells; Data; Effectiveness; Genes; Growth; Human; Human Cloning; IL2 gene; Immune; Immunoglobulin Genes; Immunotherapeutic agent; In Vitro; Interleukin-15; Interleukin-2; Knowledge; Ligands; Lymphocyte; Mediating; Modeling; Molecular; NK Cell Activation; Natural Killer Cells; Physiological; Polymerase Chain Reaction; Process; Property; Receptor Activation; Relative (related person); Role; T-Lymphocyte; Time; base; crosslink; cytokine; cytokine therapy; interest; member; programs

The main objective of this study is to determine the mechanism by which NK and T cells activated with cytokines undergo apoptosis when they encounter activating ligands on target cells. We observed that human primary NK and T cells that were stimulated in vitro with the cytokines IL2 and IL15 showed extensive potential to undergo apoptosis when their activation receptors were cross linked. In order to study this activation induced cell death in NK cells further, we drew incite from our earlier microarray study. Our microarray data on genes that are modulated upon stimulation of human NK cells in culture with IL2 showed that the antiapoptotic molecule, TOSO was down regulated in cells cultured with IL2 by 5 fold. TOSO, also called as Fas Apoptotic Inhibitory Molecule 3 (FAIM3) is a member of immunoglobulin gene superfamily and it inhibits Fas mediated apoptosis. TOSO is expressed mainly in certain classes of immune cells. However, the role of TOSO in the physiological context of NK cell activation and their function is not well known. This prompted us to look into the role of TOSO in activation induced cell death in NK cells. We also confirmed the fact that TOSO levels were down regulated and quantitated the relative levels of TOSO expression using real time PCR assays. Further, we found that human NK cells when stimulated with IL2 expressed high levels of the death receptor Fas. In order to study the involvement of TOSO in this phenomenon, we are in the process of cloning the human TOSO gene and over expressing it in primary human NK cells. We are also planning to establish a cell line model for stable expression of TOSO.

本研究的主要目的是确定被细胞因子激活的 NK 和 T 细胞在遇到靶细胞上的激活配体时发生凋亡的机制。我们观察到，在体外用细胞因子 IL2 和 IL15 刺激的人原代 NK 和 T 细胞在激活受体交联时表现出广泛的凋亡潜力。为了进一步研究这种激活诱导的 NK 细胞死亡，我们从早期的微阵列研究中汲取灵感。我们的基因微阵列数据显示，在用 IL2 培养的人 NK 细胞中刺激后受到调节的基因，抗凋亡分子 TOSO 在用 IL2 培养的细胞中下调了 5 倍。 TOSO，也称为Fas凋亡抑制分子3（FAIM3），是免疫球蛋白基因超家族的成员，它抑制Fas介导的细胞凋亡。 TOSO 主要在某些类别的免疫细胞中表达。然而，TOSO 在 NK 细胞激活的生理背景中的作用及其功能尚不清楚。这促使我们研究 TOSO 在 NK 细胞激活诱导的细胞死亡中的作用。我们还证实了 TOSO 水平下调的事实，并使用实时 PCR 测定定量了 TOSO 表达的相对水平。此外，我们发现人类 NK 细胞在受到 IL2 刺激时表达高水平的死亡受体 Fas。为了研究 TOSO 在这一现象中的作用，我们正在克隆人类 TOSO 基因并在原代人类 NK 细胞中过度表达。我们还计划建立TOSO稳定表达的细胞系模型。