Characterization of the T-cell Response to Human Norovirus Infection

摘要 慢性诺如病毒感染可导致免疫功能低下患者慢性腹泻和消瘦，包括 接受造血干细胞移植和患有原发性免疫缺陷疾病的患者。那里 目前没有治疗诺如病毒的有效疗法。我们的初步数据显示， 个体具有针对诺如病毒的T细胞免疫，并且在抗原NS6和VP1中鉴定了病毒表位 在病毒基因型中是很保守的。这项提案的首要目标是发展一部小说， 在接受HSCT的患者中治疗慢性诺如病毒感染。在我们之前的研究中，我们证明了 靶向CMV、EBV和腺病毒的病毒特异性T细胞的安全性和潜在功效以及 这种做法的可行性。了解T细胞对诺如病毒的反应可以使未来的过继性免疫试验成为可能。 NST免疫疗法，这是一种新型的抗病毒疗法，可以提供长期保护 对抗诺如病毒为了对针对诺如病毒的T细胞应答进行分类，我们现在建议从 健康供体，扩增和富集供体血液中存在的诺如病毒特异性T细胞（NST）， 通过对NST功能的广泛表征。我们假设输注NST是安全的， 有效对抗HSCT后患者的诺如病毒感染，并将恢复持久的免疫力， 诺如病毒我们进一步假设诺如病毒表位将与T细胞识别的扩增相关， 免疫显性病毒表位，其将对应于病毒基因组的稳定区域。通过这个 研究中，我们将解决以下具体目标：1）确定诺如病毒特异性T细胞是否识别 广泛数量的MHC I类和II类限制性表位，和2）为了评估诺如病毒表位是否 特异性T细胞与包括多种病毒株的广泛的分离物组交叉反应。 总的来说，这些目标将确定诺如病毒表位的范围和稳定性。完成这项研究可以 提供了一种新的抗病毒治疗，可以减少HSCT中病毒相关的发病率，并将指导未来的治疗。 针对肠道病毒的细胞疗法和疫苗试验。

Abstract Chronic norovirus infection can cause chronic diarrhea and wasting in immunocompromised patients, including those undergoing hematopoietic stem cell transplantation and with primary immunodeficiency disorders. There are currently no available therapies to treat norovirus. Our preliminary data demonstrates that healthy individuals have T cell immunity against norovirus, and that identified viral epitopes in antigens NS6 and VP1 are well conserved across viral genotypes. The overarching goal of this proposal is the development of a novel treatment for chronic norovirus infection in patients undergoing HSCT. In our previous study, we demonstrated safety and potential efficacy of virus-specific T cells targeting CMV, EBV, and adenovirus as well as the feasibility of this approach. Knowledge of T-cell response to norovirus could enable future trials of adoptive immunotherapy with NSTs, which represents a novel antiviral therapy that could provide long-term protection against norovirus. To classify the T-cell response against norovirus we now propose to take blood from the healthy donors and expand and enrich the norovirus-specific T cells (NSTs) present in donors' blood, followed by extensive characterization of the function of NSTs. We hypothesize that the infusion of NSTs will be safe and effective against norovirus infections in patients post HSCT, and will restore lasting immunity against norovirus. We further hypothesize that norovirus epitopes will correlate with expansion of T cells recognizing immunodominant viral epitopes, which will correspond to stable regions of the viral genome. Through this study, we will address the following specific aims: 1) To determine whether norovirus specific T cells recognize a broad number of MHC Class I and class II restricted epitopes, and 2) To evaluate whether norovirus epitope specific T cells are cross-reactive with a broad panel of isolates encompassing multiple viral strains. Collectively, these aims will identify the range and stability of norovirus epitopes. Completion of this study could provide a novel antiviral therapy which could reduce virus-associated morbidity in HSCT, and will guide future cellular therapy and vaccine trials targeting enteric viruses.