Recombinant life attenuated vaccine against Argentine hemorrhagic fever

描述（由申请方提供）：几种沙粒病毒在人类中引起出血热（HF）疾病，并对其流行地区的公共卫生构成严重威胁。因此，新世界沙粒病毒朱宁（JUNV）是阿根廷HF（AHF）的病原体，AHF是一种具有出血和神经系统表现的严重疾病，病死率为15- 30%。此外，JUNV非常稳定，通过气溶胶具有高度传染性，并且在低剂量下引起高发病率和死亡率，这些特征使其非常适合作为潜在的生物武器。由于美国缺乏获得许可的疫苗，以及目前的抗沙粒病毒疗法仅限于使用核苷类似物利巴韦林，因此加剧了对沙粒病毒感染的担忧，利巴韦林仅部分有效，并可引起显著毒性。因此，针对JUNV的抗病毒药物的开发被认为是HHS公共卫生紧急医疗对策企业（PHEMCE）实施计划的首要任务之一。因此，巴尔达有资格为国家战略储备采购针对JUNV感染的医疗对策。我们已经为JUNV开发了一个反向遗传系统，并完全从克隆的cDNA中拯救了感染性病毒，包括p53 d1和致病性Romero株。这一进展为我们提供了一个独特的机会，以产生一个遗传上明确定义的种子的JUNV的RJUNV/RUNV该提案旨在提供rJUNV/p53 d1cSD的稳定性、免疫原性和保护特性的全面表征，作为开发能够满足未来FDA许可所需的安全性和有效性标准的基于Candid 1的疫苗的第一步和必要步骤。为此，我们提出以下具体目标： 1.确定rJUNV/p5 d1cSD的遗传和表型稳定性。我们建议生成并表征randomd 1cSD的主病毒种子（MVS）。我们将使用来自random d1cSD的MVS在不同的细胞基质中进行独立的连续传代，包括FDA批准的疫苗生产细胞系。将对来自每个系列的选定传代的感染性后代进行遗传学（基因组测序）和表型（生长特性和毒力）表征。从这些研究中获得的遗传和表型信息将用于产生针对遗传和表型稳定性优化的新型random d1菌株。 2.确定rJUNV/rJUND 1cSD在豚鼠中的安全性、免疫原性和有效性。我们建议在豚鼠中对我们的遗传上明确定义的MVS rvd 1cSD的毒力进行全面评估。为此，我们将确定与randomd 1cSD免疫相关的临床参数，包括：体温和体重、总体活动、血液学和临床病理学数据评估（包括肾和肝功能）以及JUNV对致死性攻毒的保护。这些研究还将包括在旨在评估MVS遗传和表型稳定性的研究期间识别的相关病毒变体。 3.在普通绒猴（Callithrix jacchus）中确定rJUNV/rJUND 1cSD针对JUNV的功效。我们建议在JUNV感染的普通绒猴（Callithrix jacchus）动物模型中评估rJUNV/Rd 1cSD对由致病性rJUNV/Romero菌株引起的HF疾病的保护功效。这些研究的结果将与目标2中描述的研究获得的免疫原性数据相关。

DESCRIPTION (provided by applicant): Several arenaviruses cause hemorrhagic fever (HF) disease in humans and pose a serious threat to public health within their endemic regions. Thus, the New World arenavirus Junin (JUNV) is the etiological agent of Argentine HF (AHF), a severe illness with hemorrhagic and neurological manifestations and a case fatality of 15-30%. In addition, JUNV is very stable, highly infectious by aerosol and causes high morbidity and mortality at low dose, features that make it highly suitable as potential biological weapon. Concerns about arenavirus infections are aggravated by the lack of licensed vaccines in the US and current anti-arenavirus therapies being limited to the use of the nucleoside analogue ribavirin, which is only partially effective and can cause significant toxicity. Accordingly, the development of antiviral drugs against JUNV has been considered as one of the top priorities within the Implementation Plan of the HHS Public Health Emergency Medical Countermeasures Enterprise (PHEMCE). Therefore, medical countermeasures against JUNV infections are eligible for acquisition by BARDA for the Strategic National Stockpile. We have developed a reverse genetic system for JUNV and rescued infectious viruses, both Candid1 and the pathogenic Romero strain, entirely from cloned cDNAs. This advance has provided us with the unique opportunity to generate a genetically well-defined seeds of Candid1 (rJUNV/Candid1cSD) and Romero (rJUNV/Romero) strains of JUNV to be utilized in immunogenicity and efficacy studies. This proposal is designed to provide a comprehensive characterization of the stability, immunogenic and protective properties of rJUNV/Candid1cSD as a first and necessary step for the development of a Candid 1-based vaccine able to meet the safety and efficacy criteria required for future FDA licensing. For this we propose the following specific aims: 1. Determine the genetic and phenotypic stability of rJUNV/Candid1cSD. We propose to generate and characterize a Master Virus Seed (MVS) of rCandid1cSD. We will use the MVS from rCandid1cSD to conduct independent serial passages in different cell substrates, including FDA approved cell lines for vaccine production. Infectious progenies from selected passages from each series will be characterized genetically (genome sequencing) and phenotypically (growth properties and virulence). Genetic and phenotypic information derived from these studies will be used to generate novel rCandid1 strains optimized for genetic and phenotypic stability. 2. To determine the safety, immunogenicity and efficacy of rJUNV/Candid1cSD in guinea pigs. We propose to conduct a comprehensive assessment of the virulence of our genetically well-defined MVS rCandid1cSD in guinea pigs. For this, we will determine clinical parameters associated with immunization of rCandid1cSD including: body temperature and weight, overall activity, assessment of hematological and clinical pathology data covering kidney and liver function, and protection against lethal challenge with JUNV. These studies will also include relevant viral variants identified during studies aimed at assessing the genetic and phenotypic stability of the MVS. 3. Determine the efficacy of rJUNV/Candid1cSD against JUNV in common marmoset (Callithrix jacchus). We propose to evaluate the protective efficacy of rJUNV/Candid1cSD against HF disease caused by the pathogenic rJUNV/Romero strain in the common marmoset (Callithrix jacchus) animal model of JUNV infection. Results from these studies will be correlated with immunogenicity data obtained from studies described in aim 2.