EVALUATION OF TWO NEW CHANNENGE POOLS OF NORWALK VIRUS INOCULA IN HUMAN SUBJE

Norwalk 病毒接种物的两个新通道池在人体中的评价

• 批准号: 8166753
• 负责人: DAVID Y GRAHAM
• 金额: $ 2.37万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8166753
• 关键词: Acute; Adult; Am 80; Animal Model; Bacterial Gastroenteritis; Baculovirus Expression System; Blood; Cell Culture Techniques; Characteristics; Clinical; Clinical Research; Computer Retrieval of Information on Scientific Projects Database; Disease; Dose; Epidemic; Evaluation; Excretory function; Feces; Funding; Gastroenteritis; Genome; Goals; Grant; Hour; Human; Human Volunteers; Immune response; Immunoglobulin G; Immunoglobulins; Infection; Institution; Intestines; Life; Measures; Medicine; Methods; Molecular; Norwalk virus; Nucleic Acids; Open Reading Frames; Oral; Oral cavity; Peripheral Blood Mononuclear Cell; Phase; Preparation; Process; Proteins; Protocols documentation; Recombinants; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Safety; Saliva; Sampling; Source; Specimen; Testing; Time; Toxic effect; United States National Institutes of Health; Vaccines; Viral; Viral Antigens; Virus; Virus Shedding; Virus-like particle; Work; college; dosage; expiration; healthy volunteer; intestinal flu; meetings; prospective; prototype; response; vaccine efficacy; viral RNA

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The following is an amended submission for GCRC Protocol #2027 that incorporates changes requested by DMID/NIAID/NIH to meet FDA IND submission needs and limitations imposed by the subcontract expiration date of January 2003. Norwalk virus (NV) is the cuase of most acute , epidemic, non-bacterial gastroenteritis (24-hour intestinal flu). Although NV was identified some 30 years ago, progress in understanding its molecular characteristics has been slow due to the inability to grow the virus in cell culture and for lack of an animal model. As such, most work must be performed in human volunteers. The principal source of virus for research has been stool samples obtained from human volunteers who were experimentally infected with NV (NIH-8fIIa) obtained from the National Institutes for Health (NIH). The quantities of NV were sufficient to characterize the entire genome and significant advances have been made. The second open reading frame (ORF) of NV was expressed in the baculovirus expression system and it produced recombinant, non-infectious virus-like particles (rNV-VLPs) or protein shells that do NOT contain viral nucleic acids. A recent study by this group demonstrate that rNV-VLP preparation (a prototype vaccine), when given by mouth to human volunteers, appeared safe and universally produce an IgG immune response. It is unknown if the immune response from the prospective vaccine is protective against disease and a method to test vaccine efficacy using live virus challenge is needed. The original NV challenge pool, obtained from the NIAID/NIH, is no longer available. Using stored stool specimens obtained from an otherwise healthy subject who was infected approximately 10 years ago with NV (NIH-8fIIa), a new pool (filtrate) was developed under GMP conditions. The subject produced relatively high amounts of viral antigen and RT-PCR analysis (viral-RNA titer) of the new challenge pool (lot 42399), which was submitted by the DMID/NIAID/NIH. The safety and infectivity of the new NV challenge pool will be established by this protocol. The clinical illness rate of lot 42399 will be determined. Initally, 20 subjects will be challenged with a dose of NV (lot 42399) which approximates the dose used in previous challenge studies. The predicted rate of infection, defined by fecal viral shedding, is 80% of those who receive oral challenge. Approximately two-thirds of infected subjects are expected to become symptomatic. Subsequently, another gorup of 20 subjects will be challenged with an escalated dose to determine if the attack rate can be increased. Smaller groups of subjects will then be challenged with lower dosages to determine the Human Infectious Dose 50% (HID-50). If the initial clinical illness rate is 80% or greater, the dose escalation phase (second group of 20 subjects) will be omitted and the protocol will immediate progress to the third phase (determination of HID-50). Under this revised submission, only 42 of the orignally requested 108 subjects will be studied under method phases I and II or phases I and III, but not all three phases will be performed at this time. HYPOTHESIS A new Norwald Virus (NV) challenge pool (Lot 42399) developed and processed at Baylor College of Medicine (BCM) is safe and predictably causes a acute, self-limiting symptomatic gastroenteritis when given to healthy volunteers. The overall goal of this study is to develop a challenge pool/inoculum of Norwalk virus (NV). The specific aims and objectives of this clinical study of healthy adult subjects are: 1. To determine the safety and acute toxicity of a new NV challenge pool (Lot 42399) developed and processed at Baylor College of Medicine (BCM) in a certified Good Manufacturing Practice (cGMP) facility using Good Manufacturing Practices (GMP) 2. To determine the NV clinical attack rate induced by NV Lot 42399 3. To determine the infection rate and Human Infectious Dose 50% of NV Lot 42399. 4. To measure specific immunoglobulin responses to NV Lot 42399 inoculation/infection in blood, saliva and intestinal excretions; and 5. To examine peripheral blood mononuclear cells for the presence of NV

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 以下是GCRC方案#2027的修订提交文件，其中包含DMID/NIAID/NIH要求的变更，以满足FDA IND提交要求和2003年1月IND失效日期规定的限制。 诺瓦克病毒（NV）是最急性，流行性，非细菌性胃肠炎（24小时肠道流感）的原因。 虽然NV在大约30年前被发现，但由于无法在细胞培养中培养病毒以及缺乏动物模型，对其分子特征的理解进展缓慢。 因此，大多数工作必须在人类志愿者中进行。 用于研究的病毒的主要来源是从人类志愿者获得的粪便样品，所述人类志愿者实验性地感染了从美国国立卫生研究院（NIH）获得的NV（NIH-8fIIa）。 NV的数量足以表征整个基因组，并且已经取得了重大进展。 NV的第二个开放阅读框（ORF）在杆状病毒表达系统中表达，并且其产生重组的、非感染性病毒样颗粒（rNV-VLP）或不含病毒核酸的蛋白质壳。 该小组最近的一项研究表明，rNV-VLP制剂（一种原型疫苗），当口服给人类志愿者时，似乎是安全的，并普遍产生IgG免疫应答。 目前尚不清楚来自预期疫苗的免疫应答是否具有预防疾病的保护性，需要使用活病毒攻击来测试疫苗效力的方法。 从NIAID/NIH获得的原始NV挑战池不再可用。 使用从大约10年前感染NV（NIH-8fIIa）的健康受试者获得的储存粪便样本，在GMP条件下开发新的合并液（滤液）。 受试者产生了相对较高量的病毒抗原，并对DMID/NIAID/NIH提交的新挑战合并液（批次42399）进行了RT-PCR分析（病毒-RNA滴度）。 新NV挑战样本池的安全性和感染性将通过本方案确定。 将确定批次42399的临床患病率。 最初，20例受试者将接受NV（批次42399）激发，该剂量接近既往激发研究中使用的剂量。 通过粪便病毒脱落定义的预测感染率为接受口服攻毒的患者的80%。 预计大约三分之二的受感染者会出现症状。 随后，另一组20名受试者将接受递增剂量的挑战，以确定是否可以增加发作率。 然后将用较低剂量激发较小的受试者组，以确定人感染剂量50%（HID-50）。 如果初始临床患病率为80%或更高，则将省略剂量递增阶段（第二组20例受试者），方案将立即进展至第三阶段（确定HID-50）。 根据修订后的申报资料，最初要求的108例受试者中只有42例将在方法阶段I和II或阶段I和III进行研究，但此时不会进行所有三个阶段。 假设 在贝勒医学院（Baylor College of Medicine）开发和处理的一种新的Norwald病毒（NV）挑战池（批号42399）是安全的，并且可预测在给予健康志愿者时会引起急性自限性症状性胃肠炎。 本研究的总体目标是开发诺瓦克病毒（NV）的攻毒合并液/接种物。 本项健康成人受试者临床研究的具体目的和目标为： 1. 确定在Baylor College of Medicine（贝勒医学院）采用药品生产质量管理规范（GMP）在经认证的cGMP机构中开发和处理的新NV挑战合并液（批次42399）的安全性和急性毒性 2. 确定NV批次42399诱导的NV临床发作率 3. 确定NV批次42399的感染率和人感染剂量50%。 4. 测量血液、唾液和肠道排泄物中对NV批次42399接种/感染的特异性免疫球蛋白应答;和 5. 检测外周血单个核细胞是否存在NV