CTL-based Immunotherapy in an Organotypic Me

器官型 Me 中基于 CTL 的免疫治疗

• 批准号: 6990749
• 负责人: DOROTHEE M HERLYN
• 金额: $ 15.91万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-27 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6990749
• 关键词: T cell receptor; cytokine; cytotoxic T lymphocyte; fibroblasts; melanoma; molecular cloning; neoplasm /cancer immunology; neoplasm /cancer immunotherapy; tissue /cell culture; tumor antigens

Infiltration of primary melanomas with T lymphocytes is associated with a favorable clinical outcome. This suggests that T cell-based immunotherapies may be beneficial for melanoma patients. However, adoptive cytolytic T lymphocyte (CTL) immunotherapy of melanoma patients and active immunotherapy with CTL-based antigen (Ag)/peptide vaccines have not held promise. These limitations may rest in the selection for immunotherapy of CTL in vitro that do not reflect the Ag specificity of the CTL in situ. CTL traditionally are selected in two-dimensional mixed lymphocyte-tumor culture (MLTC) directly on plastic surfaces. We have developed the three-dimensional human melanoma organolypic culture model (melanoma reconstruct) in which CTL are cultured under tissue-like condition. Our preliminary studies suggest that the T cell receptor (TCR) repertoire in tumor reconstructs more closely resembles the repertoire of the T cells in situ as compared to the TCR repertoire in MLTC. However, the two culture systems are complementary. Thus, the chances of generating T cells with in situ (tissue)-Iike TCR are enhanced when T cells from both culture systems are cornbined, compared to either culture alone. Our major goal is to develop, including the melanoma reconstruct as a culture system, improved CTL-based adoptive and active immunotherapies against melanoma. Specifically we will: 1. Compare the phenotypic (CD markers, TCR, adhesion and apoptosis-related molecules, HLA, chemokines, chemokine receptors) and functional characteristics (proliferative and lytic activity; cytokine production) of CTL derived from fresh tissues in the reconstruct or MLTC. 2. Clone Ag(s) recognized by CTL that are highly cytolytic and express the original TCR found in situ. These Ags have vaccine potential for melanoma patients and may boost pre-existing immune responses. 3. Identify chemokines and chemokine receptors that are involved in CTL migration toward tumor cells in the reconstruct. This involves: i) Selection of migrating CTL and ii) blocking of chemokines and their receptors to determine their role in CTL migration. Chemokines relevant in CTL migration in the reconstruct may be used to enhance anti-tumor effects of adoptive and active immunotherapy. The proposed studies open new approaches to CTL-based adoptive and active immunotherapies, which may be initiated during the third year of the SPORE.

T淋巴细胞浸润原发性黑色素瘤与良好的临床结局相关。这表明基于T细胞的免疫疗法可能对黑色素瘤患者有益。然而，黑色素瘤患者的过继性溶细胞性T淋巴细胞（CTL）免疫治疗和基于CTL的抗原（Ag）/肽疫苗的主动免疫治疗并没有带来希望。这些局限性可能在于体外CTL免疫治疗的选择不反映原位CTL的Ag特异性。CTL传统上是在二维混合淋巴细胞-肿瘤培养（MLTC）中直接在塑料表面上选择的。我们建立了三维人黑色素瘤器官型培养模型（黑色素瘤重建模型）， 所述CTL在组织样条件下培养。我们的初步研究表明，与MLTC中的TCR库相比，肿瘤重建中的T细胞受体（TCR）库更接近于原位T细胞库。然而，这两种文化是互补的。因此，与单独培养相比，当来自两种培养系统的T细胞组合时，产生具有原位（组织）样TCR的T细胞的机会增加。我们的主要目标是开发，包括黑色素瘤重建作为一个培养系统，改进的CTL为基础的过继和主动免疫治疗黑色素瘤。具体来说，我们将：1。比较表型（CD标记物、TCR、粘附和骨化相关） 分子、HLA、趋化因子、趋化因子受体）和源自重建或MLTC中的新鲜组织的CTL的功能特征（增殖和裂解活性;细胞因子产生）。2. CTL识别的克隆Ag具有高度细胞溶解性并表达原位发现的原始TCR。这些抗原对黑色素瘤患者具有疫苗潜力，并可能增强预先存在的免疫反应。3.鉴定参与CTL向重建肿瘤细胞迁移的趋化因子和趋化因子受体。这涉及：i）选择迁移的CTL和ii）阻断趋化因子及其受体以确定它们在CTL迁移中的作用。重组细胞中与CTL迁移相关的趋化因子可用于增强过继免疫和主动免疫治疗的抗肿瘤效应。研究报告开启新的 基于CTL的过继性和主动免疫疗法的方法，可能在SPORE的第三年开始。