HSV Amplicon Activation of Innate and Adaptive Immunity

HSV 扩增子激活先天性和适应性免疫

• 批准号: 7081232
• 负责人: Joseph David Rosenblatt
• 金额: $ 33.06万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-23 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7081232
• 关键词: Alphaherpesvirinae; CD40 molecule; T lymphocyte; adenocarcinoma; biological signal transduction; chemokine; chronic lymphocytic leukemia; clinical research; cytotoxic T lymphocyte; gene delivery system; genetic transduction; genetically modified animals; human subject; immune tolerance /unresponsiveness; laboratory mouse; lymphoma; natural killer cells; neoplasm /cancer immunology; neoplasm /cancer immunotherapy; patient oriented research; tissue /cell culture; toll like receptor; transfection /expression vector

DESCRIPTION (provided by applicant): We are requesting continuing support for years 04-09. Our accomplishments during the previous funding period include studying the role of HSV amplicon mediated chemokine delivery alone or in combination with the co-stimulatory ligand CD40L in the murine pre B-cell tumor model A20, and the CT26 adenocarcinoma model. We demonstrated the utility of transducing tumors with the chemokines SLC alone and in combination with CD40L in eliciting recruitment of naive T-cells and dendritic cells, and facilitating development of a systemic adaptive immune response. We also demonstrated the potential utility of helper virus free HSV amplicons for transduction of human lymphoid malignancies. HSV amplicons encoding chemokines and/or costimulatory ligands elicit an unusually vigorous response resulting in eradication of multicentric tumors and the establishment of long lasting immunity. This suggests a particularly productive interplay between the innate and adaptive immune systems. We propose to continue studies of HSV amplicon mediated immunotherapy in murine tumors and human lymphoid tumors, with new emphasis on the role of the innate immune response. The overall goal is to understand the nature of the innate response to HSV amplicon transduction, and the role of the innate response in priming a subsequent adaptive response. Aim I, will focus on HSV amplicon engagement of the innate immune system. The chemokine/cytokine profile generated in response to vector transduction in normal splenocytes in vitro will be studied. We will study TLR receptor signaling in response to HSV amplicons. In Aim II we will compare the anti-tumor response mediated by SLC delivered using HSV amplicon vectors to that seen with nonviral delivery systems. Next we will study the effect of HSV amplicons encoding effector molecules that will selectively recruit and activate the innate system on anti-tumor response. HSV amplicons encoding chemokines that can recruit NK cells and/or NKG2D ligands will be delivered to augment the innate response in the mouse tumor models CT26, and A20. Using Ovalbumin expressing EG.7 cells, and OVA specific TCR transgenic T-cells from OT-1 mice, we will study how signals from within the innate system impact subsequent development of an adaptive response. Mice with genetic defects in key components of the innate response and the innate/adaptive transition will be tested for ability to support OT-1 activation and expansion following amplicon transduction. In Aim III we will study HSV amplicon capacity to induce NKG2D ligands on human Chronic Lymphocytic Leukemia (CLL) cells. We will also assay effects of direct transduction using HSV amplicons encoding NKG2D ligands on the immunogenicity of CLL cells. Insights gained from murine and human studies proposed will afford a better understanding of the innate contribution to development of an adaptive anti tumor response, and may lead to novel clinical approaches.

描述（由申请人提供）：我们要求在04-09年继续提供支持。我们在上一个资金期间的成就包括研究HSV扩增子介导的趋化因子递送的作用，或与鼠前B细胞肿瘤模型A20和CT26腺癌模型中的共刺激配体CD40L结合使用。我们证明了单独使用趋化因子SLC转导肿瘤并与CD40L结合的实用性，从而引发了幼稚的T细胞和树突状细胞的募集，并促进了系统性适应性免疫反应的发展。我们还证明了无辅助病毒无辅助性HSV扩增子对人淋巴恶性肿瘤转导的潜在效用。编码趋化因子和/或共刺激配体的HSV扩增子引起了异常剧烈的反应，导致根除多中心肿瘤并建立了持久的免疫力。这表明先天和适应性免疫系统之间的效率特别有效。我们建议继续研究HSV扩增子在鼠肿瘤和人淋巴肿瘤中介导的免疫疗法，并有了新的重视先天免疫反应的作用。总体目标是了解先天对HSV扩增子转导的响应的性质，以及先天反应在启动随后的自适应反应中的作用。 AIM I将重点放在先天免疫系统的HSV Amplicon参与度上。将研究响应于正常脾细胞的载体转导而产生的趋化因子/细胞因子谱。我们将根据HSV扩增子研究TLR受体信号传导。 在AIM II中，我们将比较使用HSV Amplicon向量传递的SLC介导的抗肿瘤反应与非病毒输送系统所见的抗肿瘤反应。接下来，我们将研究编码效应子分子的HSV扩增子的效果，这些分子将有选择地募集和激活先天系统对抗肿瘤反应。编码可以募集NK细胞和/或NKG2D配体的趋化因子的HSV扩增子将被递送以增强小鼠肿瘤模型CT26和A20中的先天反应。使用表达EG.7细胞的卵蛋白和OVA特异性TCR转基因T细胞，我们将研究先天系统内的信号如何影响随后的自适应反应的发展。具有先天反应的关键成分和先天/适应性转变的关键组成部分中具有遗传缺陷的小鼠，将测试在扩增子转导后支持OT-1激活和扩张的能力。 在AIM III中，我们将研究HSV扩增子的能力，以诱导人类慢性淋巴细胞性白血病（CLL）细胞诱导NKG2D配体。我们还将使用编码NKG2D配体对CLL细胞的免疫原性的HSV扩增子进行直接转导的影响。 从鼠和人类研究中获得的见解将更好地理解对自适应抗肿瘤反应发展的先天贡献，并可能导致新颖的临床方法。