Optimizing Induction Conditions for Immunotherapeutic CTL

描述（由申请人提供）：该项目的目标是：1)开发带有模式识别受体（PRR）分子的生物相容性纳米颗粒（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效应细胞的产生和功能。4)在GL261小鼠胶质瘤模型中评估同种异体反应性CTL和自体肿瘤相关抗原（TAA）定向CTL，肿瘤细胞接种物对脑肿瘤干细胞（BTSC）富集或不富集。后一个目标将允许确定BTSC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、LPS和甘油三酯化BSA）作为生物相容性PLGA载体基质的成分，这些成分已被批准用于临床，以刺激DC达到可变激活状态。在一些实验中，复杂的NP-PRR还包含肿瘤相关抗原（mEphA2, hgp100， mTRP-2和GARC-1）“使用来自未成熟骨髓（BM）的传统和Flt3配体DC作为前体细胞”，NP-PRR激活的DC将被用来刺激同种反应性或自体幼稚T细胞来效应细胞毒性T淋巴细胞（CTL）“NP-PRR刺激的DC和随后刺激的CTL将在体外功能和表型上进行表征。”受刺激的同种异体反应性CTL或taa定向CTL在过继转移到血清培养的GL261或神经球培养的对应物小鼠后，其抗肿瘤作用将在功能和表型上具有特征。

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.