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(申请人提供)：我们请求在04-09年度继续提供支持。在之前的资助期间，我们的成就包括研究单纯疱疹病毒扩增子介导的趋化因子传递在小鼠前B细胞肿瘤模型A20和CT26腺癌模型中的作用，或者联合共刺激配体CD40L的作用。我们证明了单独使用趋化因子SLC转导肿瘤以及与CD40L联合使用在诱导幼稚T细胞和树突状细胞的募集以及促进全身适应性免疫反应的发展方面的效用。我们还证明了无辅助病毒HSV扩增在转导人类淋巴系统恶性肿瘤方面的潜在用途。HSV编码趋化因子和/或共刺激配体的扩增片段能引起异常强烈的反应，导致多中心肿瘤的根除和持久免疫的建立。这表明先天免疫系统和获得性免疫系统之间存在着一种特别富有成效的相互作用。我们建议继续研究HSV扩增子介导的免疫治疗在小鼠肿瘤和人类淋巴肿瘤中的作用，并重新强调先天免疫反应的作用。总体目标是了解对HSV扩增子转导的先天反应的性质，以及先天反应在启动随后的适应性反应中的作用。目的I，将重点放在HSV扩增子参与的先天免疫系统。在体外，我们将研究正常脾细胞在载体转导下产生的趋化因子/细胞因子的分布。我们将研究TLR受体信号对HSV扩增的反应。 在AIM II中，我们将比较使用HSV扩增载体和非病毒传递系统传递的SLC介导的抗肿瘤反应。接下来，我们将研究编码效应分子的单纯疱疹病毒扩增物在抗肿瘤反应中的作用，该效应分子将选择性地招募和激活固有系统。HSV编码能够招募NK细胞和/或NKG2D配体的趋化因子的扩增片段将被递送到小鼠肿瘤模型CT26和A20中以增强先天反应。使用表达卵清蛋白的EG.7细胞和来自OT-1小鼠的OVA特异性TCR转基因T细胞，我们将研究来自固有系统内的信号如何影响随后的适应性反应的发展。在先天反应和先天/适应性转换的关键成分中存在遗传缺陷的小鼠，将在扩增载体转导后接受支持OT-1激活和扩张的能力测试。 在目标III中，我们将研究HSV扩增子在人慢性淋巴细胞白血病(CLL)细胞上诱导NKG2D配体的能力。我们还将检测使用编码NKG2D配体的HSV扩增直接转导对CLL细胞免疫原性的影响。 从小鼠和人类研究中获得的见解将使我们更好地理解适应性抗肿瘤反应的先天贡献，并可能导致新的临床方法。