Optimizing Induction Conditions for Immunotherapeutic CTL

优化免疫治疗 CTL 的诱导条件

基本信息

批准号：
8307525
负责人：
CAROL A KRUSE
金额：
$ 39.44万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-13 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8307525
关键词：
Adoptive Transfer Affect Agonist Antigen-Presenting Cells Antigens Apoptosis Autoantigens Autologous Biocompatible Biological Assay Bone Marrow Brain Neoplasms C Type Lectin Receptors C57BL/6 Mouse Cancer Vaccines Cell Culture Techniques Cells Characteristics Clinical Clinical Research Complex Culture Media Cytolysis Cytotoxic T-Lymphocytes DNA Dendritic Cell Vaccine Dendritic Cells Dendritic cell activation Effector Cell Elements Generations Glioma Goals Imiquimod Immature Bone Immunologist Immunotherapeutic agent Immunotherapy In Vitro Individual Letters Ligands Light Lymphocyte MHC Class I Genes Measures Mixed Neoplasm Modeling Morphology Mus Nature Particle Size Pattern recognition receptor Peptides Preparation Production Radio Reaction Research Research Design Research Personnel Screening procedure Serum Signaling Molecule Splenocyte Staining method Stains Stimulus Surface T-Lymphocyte Techniques Testing Therapeutic Uses Toll-like receptors Tumor Antigens Tumor Stem Cells Vaccines Work cancer immunotherapy cancer therapy carcinogenesis cell injury clinical application computerized cytokine cytotoxic cytotoxicity experience in vivo interest mouse model nanomedicine nanoparticle neoplastic cell pathogen precursor cell public health relevance research study response stem cell therapy success therapy resistant translational medicine tumor

项目摘要

DESCRIPTION (provided by applicant): The project goals are 1) to develop biocompatible nanoparticles (NP) bearing Pattern Recognition Receptor (PRR) molecules, i.e., agonists for Toll Like Receptors or C-type Lectin Receptors, 2) to determine if these NP- PRR differently affect activation of dendritic cells (DC), 3) to determine if the differently activated DC then influence the generation and functionality of T effector cells, and 4) to evaluate the alloreactive CTL and the autologous tumor associated antigen (TAA)-directed CTL in the GL261 mouse glioma model, where the tumor cell inoculum either is or is not enriched for brain tumor stem cells (BTSC). The latter goal will allow for a determination of BTSC immunosensitivity/immunoresistance. The cancer immunotherapy approach will: "Use various NP-PRR (CpG DNA, imiquimod, LPS and mannosylated BSA) that will be components of a biocompatible PLGA carrier matrix already approved for clinical use to stimulate DC to variable activation states. In some experiments complex NP-PRR will also contain tumor associated antigens (mEphA2, hgp100, mTRP-2 and GARC-1) " Use conventional and Flt3 ligand DC derived from immature bone marrow (BM) as precursor cells " NP-PRR activated DC will then be used to stimulate alloreactive or autologous naive T cells to effector cytotoxic T lymphocytes (CTL) "NP-PRR stimulated DC and subsequently stimulated CTL will be functionally and phenotypically characterized in vitro."Stimulated alloreactive CTL or TAA-directed CTL will be functionally and phenotypically characterized for their antitumor effects after their adoptive transfer into mice bearing serum-cultured GL261 or the neurosphere-cultured counterpart. PUBLIC HEALTH RELEVANCE: The work proposed here is an interdisciplinary project in translational medicine that brings together several key elements: 1. Cytotoxic T Lymphocytes (CTL)-effector cells for immunotherapy 2. Dendritic Cells (DC)-antigen presenting cells (APC) used to induce CTL effectors in vitro 3. Nanoparticles (NP) and their payload-activation stimuli and antigens delivered to DC 4. Brain Tumor Stem Cells (BTSC)-therapy-resistant tumor cells that should be included as targets of the effector cells The goal of this project is to modulate dendritic cell activation with pathogen associated signaling molecules contained with nanoparticles. These nanoparticles will be used to stimulate dendritic cells to various activation states. The dendritic cells will then be used to stimulate naive T cells into effector cells. The generated CTL will be functionally and phenotypically characterized in vitro and in vivo using mouse glioma cell targets or those enriched for brain tumor stem cells in culture.

DESCRIPTION (provided by applicant): The project goals are 1) to develop biocompatible nanoparticles (NP) bearing Pattern Recognition Receptor (PRR) molecules, i.e., agonists for Toll Like Receptors or C-type Lectin Receptors, 2) to determine if these NP- PRR differently affect activation of dendritic cells (DC), 3) to determine if the differently activated DC then influence the generation and functionality of T效应细胞和4）在GL261小鼠胶质瘤模型中评估同种反应性CTL和自体肿瘤相关的抗原（TAA）指导的CTL，其中肿瘤细胞接种物是或不富含脑肿瘤干细胞（BTSC）的肿瘤细胞。后一个目标将允许确定BTSC免疫敏感/免疫阻力。癌症免疫疗法方法将：“使用各种NP-PRR（CPG DNA，咪喹莫德，LPS和甘露糖基化的BSA），这将是生物相容性的PLGA载体基质的组成部分，该载体矩阵已批准已经促进了临床用途，以刺激DC到可变激活状态。 GARC-1) " Use conventional and Flt3 ligand DC derived from immature bone marrow (BM) as precursor cells " NP-PRR activated DC will then be used to stimulate alloreactive or autologous naive T cells to effector cytotoxic T lymphocytes (CTL) "NP-PRR stimulated DC and subsequently stimulated CTL will be functionally and phenotypically characterized in “刺激的同种反应性CTL或TAA指导的CTL将在功能和表型上以其抗肿瘤作用的特征在于它们的抗肿瘤效应，它们的收养转移到带有血清培养的GL261或神经圈培养的小鼠中。 PUBLIC HEALTH RELEVANCE: The work proposed here is an interdisciplinary project in translational medicine that brings together several key elements: 1. Cytotoxic T Lymphocytes (CTL)-effector cells for immunotherapy 2. Dendritic Cells (DC)-antigen presenting cells (APC) used to induce CTL effectors in vitro 3. Nanoparticles (NP) and their payload-activation stimuli and递送到DC 4的抗原。脑肿瘤干细胞（BTSC） - 耐药性肿瘤细胞应作为效应细胞的靶标，该项目的目标是通过纳米颗粒包含的病原体信号分子调节树突状细胞激活。这些纳米颗粒将用于刺激各种激活态的树突状细胞。然后，树突状细胞将用于将幼稚的T细胞刺激到效应细胞中。生成的CTL将在功能和表型上使用小鼠胶质瘤细胞靶标的在体外和体内表征，或在培养中富含脑肿瘤干细胞的CTL。