LEUKOCYTE-ENDOTHELIAL CELL ADHESION IN TUMOR IMMUNITY

肿瘤免疫中的白细胞-内皮细胞粘附

• 批准号: 6329055
• 负责人: Sharon S Evans
• 金额: $ 20.15万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-12-13 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6329055
• 关键词: T cell receptor; antineoplastics; biological signal transduction; cell adhesion; cytotoxic T lymphocyte; disease /disorder model; genetically modified animals; hyperthermia therapy; immunocytochemistry; integrins; interleukin 6; laboratory mouse; leukocytes; lymph nodes; monoclonal antibody; natural killer cells; neoplasm /cancer immunology; neoplastic cell; neoplastic process; pancreatic islet neoplasm; protein structure function; receptor expression; selectins; tissue /cell culture; vascular endothelium

This proposal addresses the central hypothesis that significant antitumor responses can be achieved through the use of fever- range hyperthermia to stimulate L-selectin and alpha4beta7 integrin-dependent recruitment of immune effector cells to malignant tissues and tumor-draining lymph nodes. The hypothesis is formulated on the basis of new information demonstrating that fever range, long duration whole body hyperthermia (WBH) stimulates expression of endothelial ligands for two leukocyte homing receptors, L-selectin and alpha4beta7 integrin, on tumor vessels in the RIP-Tag5 transgenic mouse model. This finding is correlated with increased lymphocyte infiltration into beta islet tumors. Notably, pancreatic tumors that develop as a result of transgene expression of the SV40 T antigen (Tag) in RIP-Tag5 mice are normally devoid of infiltrating lymphocytes. Three independent approaches are proposed to address the central hypothesis: (1) Strategies are designed to examine the direct role of L-selectin and alpha4beta7 integrin in targeting immune effector cells to pancreatic beta islet tumors in response to fever-range WBH in RIP-Tag5 single transgenic mice and in RIP- Tag5 double transgenic mice that have a high frequency of tumor- reactive cytotoxic T lymphocytes (CTL) as a result of coexpression of the B7.1 costimulatory molecule or a Tag-specific T cell receptor. The specific role of adhesion molecules in lymphocyte recruitment will be delineated by antibody blockade studies and using cell lines that restrictively bind to endothelium via L-selectin or alpha4beta7 integrin. (2) The molecular mechanisms underlying hyperthermia control of lymphocyte-endothelial adhesion will be investigated in cell lines using in vitro kinase assays and dominant negative approaches to examine the involvement of specific signal transduction pathways (i.e., MAPK, HSP90, JAK/STATs, Src kinases, and IL-6). (3) Studies are designed to determine if fever-range WBH, in combination with cytokines (IFN-alpha, IL-2), inhibits tumor progression in RIP-Tag transgenic mice by enhancing L- selectin and/or alpha4beta7 integrin-dependent accumulation of CTL in beta islet tumors. WBH effects on intratumoral localization of immune cells that produce Th1-cytokines (e.g., IFN-gamma) will also be examined since these cytokines are fundamental to the generation of cell-mediated tumor immunity. The proposed studies are expected to provide a framework for future evaluation of the efficacy of fever-range WBH as an adjuvant in the treatment of cancer in combination with immunotherapies designed to enhance the generation of tumor- specific CTL.

该提议解决了中心假设，即通过使用发热范围的高热来刺激L-选择素和α 4 β 7整联蛋白依赖性的免疫效应细胞向恶性组织和肿瘤引流淋巴结的募集，可以实现显著的抗肿瘤应答。 新的信息表明，发热范围，长时间全身热疗（WBH）刺激表达的内皮配体的两个白细胞归巢受体，L-选择素和α 4 β 7整联蛋白，在RIP-Tag 5转基因小鼠模型的肿瘤血管的基础上制定的假设。 这一发现与β胰岛肿瘤中淋巴细胞浸润增加相关。 值得注意的是，在RIP-Tag 5小鼠中由于SV 40 T抗原（Tag）的转基因表达而发展的胰腺肿瘤通常缺乏浸润性淋巴细胞。 提出了三种独立的方法来解决中心假设：（1）设计策略以检查L-选择素和α 4 β 7整联蛋白在RIP-Tag 5单转基因小鼠和RIP-Tag 5双转基因小鼠中响应于发热范围WBH将免疫效应细胞靶向胰岛β肿瘤中的直接作用，所述RIP-Tag 5单转基因小鼠和RIP-Tag 5双转基因小鼠具有高频率的肿瘤反应性细胞毒性T淋巴细胞（CTL）。作为B7.1共刺激分子或标签特异性T细胞受体共表达的结果。 粘附分子在淋巴细胞募集中的特定作用将通过抗体阻断研究和使用通过L-选择素或α 4 β 7整联蛋白限制性结合内皮的细胞系来描述。 (2)将使用体外激酶测定和显性阴性方法在细胞系中研究淋巴细胞-内皮粘附的高温控制的分子机制，以检查特定信号转导途径的参与（即，MAPK、HSP 90、JAK/STAT、Src激酶和IL-6）。 (3)设计研究以确定发热范围的WBH与细胞因子（IFN-α、IL-2）组合是否通过增强β胰岛肿瘤中CTL的L-选择素和/或α 4 β 7整联蛋白依赖性积累来抑制RIP-Tag转基因小鼠中的肿瘤进展。 WBH对产生Th 1-细胞因子（例如，IFN-γ），因为这些细胞因子是细胞介导的肿瘤免疫产生的基础。预期所提出的研究将为未来评估发热范围WBH作为佐剂与旨在增强肿瘤特异性CTL产生的免疫疗法组合治疗癌症的功效提供框架。