Therapeutic targets in gammaherpesvirus infection

项目摘要 人类γ疱疹病毒（γ HV）、卡波西肉瘤相关疱疹病毒和爱泼斯坦-巴尔病毒是 DNA肿瘤病毒建立终身感染。这些病毒与致病性 免疫抑制的结果，包括艾滋病定义的恶性肿瘤卡波西肉瘤和非艾滋病定义的恶性肿瘤卡波西肉瘤。 霍奇金淋巴瘤。γ HV的一个主要挑战是，这些病毒存在病毒再感染的终身风险。 出现和发病机制，特别是在获得性或医学诱导的免疫抑制的情况下。 针对γHV疾病的有限治疗干预进一步混淆了终身感染。到目前为止， 对γ HV诱导的疾病的最佳防御是完整的免疫系统。尽管存在这种相关性， 在我们对以下问题的理解中仍然存在主要的知识差距：1）成功接种γHV的保护性方面 感染，和2）疫苗接种对感染细胞的组成和频率的后果，无论是在 或在感染后。在R 01申请中，我们试图研究疫苗接种如何改变γHV 感染，使用鼠γ疱疹病毒68（γ HV 68），一种γHV感染的小鼠模型，有助于研究 感染和相关反应，从原发感染到终身潜伏期和再次出现溶解性 感染我们最近进行了研究，以表征潜伏期如何受到病毒和宿主因素的调节， 单细胞水平，基于潜伏期的表达识别潜伏期池是异质的， 相关核抗原（拉娜），并且拉娜+潜伏感染细胞的比例受到 调控这些研究进一步确定了CD 8 T细胞和细胞因子干扰素-γ限制了拉娜+ 潜伏感染的B细胞，并且用减毒活疫苗接种能够深刻地限制 拉娜+潜伏性B细胞感染以干扰素-γ-非依赖性方式。我们现在寻求调查 骨髓细胞在感染中的作用和疫苗接种对γHV感染的影响我们将利用 使用单细胞免疫技术确定关键特征的疫苗接种方案， 分析和高维方法来区分疫苗效力的特征。我们假设 用减毒活疫苗接种诱导：1）局部和全身骨髓重编程， 其2）将γHV骨髓感染重定向到免疫敏感的骨髓靶。我们将在 三个目标。首先，我们将分析疫苗接种对局部和全身骨髓细胞的影响。二是明确 疫苗接种如何影响骨髓细胞的原发性γHV感染。第三，剖析病毒的嗜性是如何受到 疫苗诱导的效应机制。通过分析特定的病毒和宿主过程， 疫苗具有不同的功效，这项建议旨在研究骨髓重编程作为一种必要的， 治疗上可行的针对γHV潜伏期的疫苗接种的靶点。

PROJECT SUMMARY The human gammaherpesviruses (γHVs) Kaposi's sarcoma associated herpesvirus and Epstein-Barr virus are DNA tumor viruses that establish a lifelong infection. These viruses are strongly associated with pathogenic outcomes in immune suppression, including the AIDS defining malignancies Kaposi's sarcoma and non- Hodgkin lymphoma. A major challenge of the γHVs is that these viruses present a lifelong risk for viral re- emergence and pathogenesis, especially in the context of acquired or medically-induced immunosuppression. Lifelong infection is further confounded by the limited therapeutic interventions against γHV diseases. To date, the best defense against γHV-induced disease is an intact immune system. Despite this correlation, there remain major knowledge gaps in our understanding of: 1) protective aspects of successful vaccination to γHV infection, and 2) consequence of vaccination on the composition and frequency of infected cells, either before or after established infection. In this R01 application, we seek to investigate how vaccination alters γHV infection, using murine gammaherpesvirus 68 (γHV68), a mouse model of γHV infection, that facilitates study of infection and associated responses from primary infection through lifelong latency and re-emergence of lytic infection. We recently undertook studies to characterize how latency is regulated by viral and host factors at the single-cell level, identifying that the latent pool is heterogeneous based on expression of the latency- associated nuclear antigen (LANA), and that the proportion of LANA+ latently-infected cells is subject to regulation. These studies further identified that CD8 T cells, and the cytokine interferon-gamma, limit LANA+ latently-infected B cells, and that vaccination with a live-attenuated vaccine is capable of profoundly restricting LANA+ latent B cell infection in an interferon-gamma-independent manner. We now seek to investigate the role of myeloid cells in infection and vaccination on γHV infection both in vitro and in vivo. We will make use of vaccination protocols with defined differences in efficacy to identify the critical features using single cell analysis and high dimensional methods to distinguish the features of vaccine efficacy. We hypothesize that vaccination with a live-attenuated vaccine induces: 1) local and systemic myeloid reprogramming, which 2) redirects γHV myeloid infection into an immune-susceptible myeloid target. We will test this in three aims. First, we will analyze the impact of vaccination on local and systemic myeloid cells. Second, define how vaccination affects primary γHV infection of myeloid cells. Third, dissect how viral tropism is influenced by vaccine-induced effector mechanisms. By analyzing specific viral and host processes in the context of vaccines with varying efficacy, this proposal seeks to investigate myeloid reprogramming as a necessary, and therapeutically viable, target for vaccination against γHV latency.