Optimizing Induction Conditions for Immunotherapeutic CTL

项目描述(由申请人提供)：该项目的目标是1)开发含有模式识别受体(PRR)分子的生物相容纳米颗粒(NP)，即Toll样受体或C型凝集素受体的激动剂；2)确定这些NP-PRR是否会以不同的方式影响树突状细胞(DC)的激活；3)确定不同激活的DC是否会影响T效应细胞的生成和功能；4)评估GL261小鼠脑胶质瘤模型中同种异体反应性CTL和自体肿瘤相关抗原(TAA)导向的CTL，其中肿瘤细胞接种是或不是富集脑肿瘤干细胞(BTSC)。后一个目标将允许确定BTSC的免疫敏感性/免疫耐药性。癌症免疫治疗方法将：“使用各种NP-PRR(CpG DNA、咪喹莫特、脂多糖和甘露糖化BSA)，这些将是已被批准用于临床的生物兼容PLGA载体基质的组成部分，以刺激DC进入不同的激活状态。在一些实验中，复杂的NP-PRR还将包含肿瘤相关抗原(mEphA2，hgp100，mTRP-2和GARC-1)“使用来自未成熟骨髓(BM)的常规和Flt3配体DC作为前体细胞”NP-PRR激活的DC将被用来刺激同种异体反应性或自体初始T细胞来影响细胞毒性T淋巴细胞(CTL)“NP-PRR刺激的DC和随后被刺激的CTL将在体外进行功能和表型鉴定。”被刺激的同种异体反应性CTL或TAA导向的CTL将在过继转移到带有血清培养的GL261或神经球培养的对应物的小鼠体内后，在功能和表型上表征它们的抗肿瘤效果。

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.