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.

这项研究的主要目的是确定NK和T细胞在遇到靶细胞上激活配体时通过细胞因子激活凋亡的机制。我们观察到，在体外与细胞因子IL2和IL15体外刺激的人初级NK和T细胞在交叉链接时具有广泛的受体凋亡的潜力。为了研究这种激活诱导NK细胞中的细胞死亡，我们从较早的微阵列研究中汲取了煽动。我们对使用IL2培养的人NK细胞调节基因的微阵列数据表明，在用IL2培养的细胞中调节抗凋亡分子，TOSO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，它抑制了FAS介导的凋亡。 TOSO主要在某些类型的免疫细胞中表达。然而，TOSO在NK细胞激活及其功能的生理环境中的作用尚不清楚。这促使我们研究了TOSO在激活诱导NK细胞中的细胞死亡中的作用。我们还证实了这样一个事实，即使用实时PCR分析，对TOSO水平下调并定量TOSO表达的相对水平。此外，我们发现用IL2刺激的人NK细胞表达高水平的死亡受体FAS。为了研究TOSO在这种现象中的参与，我们正在克隆人类TOSO基因并在原代人NK细胞中表达它。我们还计划建立一个细胞系模型，以稳定地表达TOSO。