Characterization of the T-cell Response to Human Norovirus Infection

摘要 慢性诺沃克病毒感染可导致免疫功能低下患者的慢性腹泻和消瘦，包括 接受造血干细胞移植和患有原发免疫缺陷疾病的患者。那里 目前还没有治疗诺沃克病毒的可用的疗法。我们的初步数据表明，健康 个体对诺如病毒有T细胞免疫力，并在NS6和VP1抗原中识别出病毒表位 在不同的病毒基因型中都保存得很好。这项提议的首要目标是发展一部小说 造血干细胞移植患者慢性诺如病毒感染的治疗。在我们之前的研究中，我们展示了 针对CMV、EBV和腺病毒的病毒特异性T细胞的安全性和潜在有效性以及 这一方法的可行性。了解T细胞对诺沃克病毒的反应可能使未来的过继试验成为可能 NSTs免疫疗法，这代表了一种新的抗病毒疗法，可以提供长期保护 对抗诺沃克病毒。为了对诺沃克病毒的T细胞反应进行分类，我们现在建议从 健康捐赠者并扩增和丰富捐赠者血液中存在的诺沃克病毒特异性T细胞(NST)，随后 通过对NST的功能进行广泛的表征。我们假设输注NSTs将是安全的 对造血干细胞移植后患者的诺沃克病毒感染有效，并将恢复对 诺沃克病毒。我们进一步假设诺如病毒表位将与T细胞识别的扩大相关 免疫优势的病毒表位，将与病毒基因组的稳定区域相对应。通过这件事 在这项研究中，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 广泛的MHC I类和II类限制性表位，以及2)评估诺如病毒表位 特定的T细胞与包括多种病毒株的广泛分离株发生交叉反应。 总的来说，这些目标将确定诺沃克病毒表位的范围和稳定性。完成这项研究可能会 提供了一种新的抗病毒治疗方法，可以减少HSCT中病毒相关的发病率，并将指导未来 针对肠道病毒的细胞治疗和疫苗试验。

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.