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: 6050909
• 负责人: Sharon S Evans
• 金额: $ 19.97万
• 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/6050909
• 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.

该提案提出了一个中心假设，即通过使用发烧范围的热疗来刺激 L-选择素和 α4β7 整合素依赖的免疫效应细胞向恶性组织和肿瘤引流淋巴结的募集，可以实现显着的抗肿瘤反应。 该假设是基于新的信息提出的，该新信息表明，发烧范围、长时间的全身热疗（WBH）会刺激 RIP-Tag5 转基因肿瘤血管上两种白细胞归巢受体（L-选择素和 α4β7 整合素）的内皮配体的表达鼠标模型。 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，RIP-Tag5 小鼠中因 SV40 T 抗原 (Tag) 转基因表达而产生的胰腺肿瘤通常缺乏浸润淋巴细胞。 提出了三种独立的方法来解决中心假设：（1）设计策略来检查L-选择素和α4β7整合素在将免疫效应细胞靶向胰腺β岛肿瘤以响应RIP-Tag5中的发烧范围WBH中的直接作用单转基因小鼠和 RIP-Tag5 双转基因小鼠由于 B7.1 共刺激的共表达而具有高频率的肿瘤反应性细胞毒性 T 淋巴细胞 (CTL)分子或标签特异性 T 细胞受体。 粘附分子在淋巴细胞募集中的具体作用将通过抗体阻断研究和使用通过 L-选择素或 α4β7 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失活方法在细胞系中研究淋巴细胞-内皮粘附的热控制的分子机制，以检查特定信号转导途径（即 MAPK、HSP90、JAK/STATs、Src）的参与。激酶和 IL-6）。 (3) 研究旨在确定发烧范围的 WBH 与细胞因子（IFN-α、IL-2）结合是否通过增强 L-选择素和/或 α4β7 整合素依赖性积累来抑制 RIP-Tag 转基因小鼠的肿瘤进展β 岛肿瘤中 CTL 的作用。 还将检查 WBH 对产生 Th1 细胞因子（例如 IFN-gamma）的免疫细胞瘤内定位的影响，因为这些细胞因子是产生细胞介导的肿瘤免疫的基础。拟议的研究预计将为未来评估发烧范围 WBH 作为癌症治疗佐剂与旨在增强肿瘤特异性 CTL 生成的免疫疗法相结合的功效提供一个框架。