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: 7605923
• 负责人: DAVID Y GRAHAM
• 金额: $ 12.89万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-15 至 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7605923
• 关键词: Acute; Adult; Am 80; Animal Model; Bacterial Gastroenteritis; Baculovirus Expression System; Blood; Characteristics; Clinical; Clinical Research; Computer Retrieval of Information on Scientific Projects Database; Condition; Cultured Cells; 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; Rate; Recombinants; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Safety; Saliva; Sampling; Source; Specimen; Testing; Time; Toxic effect; United States Food and Drug Administration; United States National Institutes of Health; Vaccines; Viral; Viral Antigens; Virus; Virus Shedding; Virus-like particle; Work; abstracting; college; dosage; expiration; healthy volunteer; intestinal flu; 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. ABSTRACT 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. SPECIFIC AIMS 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利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 摘要 以下是GCRC第#2027号议定书的修订申请，其中纳入了DMID/NIAID/NIH要求的更改，以满足FDA IND提交的需要和2003年1月分包合同到期日施加的限制。 诺沃克病毒(NV)是大多数急性、流行性、非细菌性胃肠炎(24小时肠道流感)的病因。尽管NV在大约30年前就被发现，但由于无法在细胞培养中培养病毒，以及缺乏动物模型，对其分子特征的了解进展缓慢。因此，大多数工作必须在人类志愿者身上完成。用于研究的病毒的主要来源是从从国家卫生研究院(NIH)获得的实验感染NV(NIH-8FIIa)的人类志愿者那里获得的粪便样本。NV的数量足以描述整个基因组的特征，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，它产生不含病毒核酸的重组非传染性病毒样颗粒(RNV-VLP)或蛋白外壳。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，并普遍产生免疫球蛋白免疫反应。目前尚不清楚来自预期疫苗的免疫反应是否对疾病具有保护作用，因此需要一种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。利用从一名大约10年前感染NV(NIH-8FIIa)的健康受试者那里获得的储存粪便样本，在GMP条件下开发了一种新的池(滤液)。受试者产生了相对较高数量的病毒抗原，并对新的挑战池(批次42399)进行了RT-聚合酶链式反应分析(病毒-RNA滴度)，这是由DMID/NIAID/NIH提交的。新的NV挑战池的安全性和传染性将通过该协议建立。42399号拍品的临床患病率将确定。最初，20名受试者将接受NV(批次42399)的挑战，剂量与以前挑战研究中使用的剂量大致相同。根据粪便病毒脱落的定义，预计感染率为接受口服挑战的人的80%。预计大约三分之二的感染者会出现症状。随后，另一组20名受试者将接受挑战，增加剂量，以确定是否可以增加发病率。然后，较小的受试组将接受较低剂量的挑战，以确定人类感染剂量的50%(HID-50)。如果初始临床发病率为80%或更高，剂量递增阶段(第二组20名受试者)将被省略，方案将立即进入第三阶段(HID-50测定)。根据这份订正提交的材料，最初要求的108个受试者中只有42人将在方法第一阶段和第二阶段或第一阶段和第三阶段进行研究，但不是所有三个阶段都将在此时进行。 假设 贝勒医学院开发和加工的一种新的诺瓦尔德病毒(NV)攻击池(批号42399)是安全的，当给健康志愿者服用时，可以预测会导致急性自限性症状性胃肠炎。 具体目标 这项研究的总体目标是开发诺瓦克病毒(NV)的挑战池/疫苗。 这项针对健康成人受试者的临床研究的具体目的和目标是： 1.确定在贝勒医学院开发和处理的新NV挑战池(批次42399)的安全性和急性毒性，该池在使用良好制造规范的认证良好制造规范(CGMP)设施中进行处理 2.NV批次42399诱发NV临床发作率的测定 3.测定42399批次新城疫病毒的感染率和人感染剂量。 4.检测血液、唾液和肠道排泄物对新城疫42399批疫苗接种/感染的特异性免疫球蛋白反应；以及 5.检查外周血单个核细胞是否存在新城疫