Human Mab Cocktails to Prevent & Treat H5N1 Avian Influenza

人类单克隆抗体鸡尾酒可预防

• 批准号: 7668571
• 负责人: Wayne A. Marasco
• 金额: $ 130.97万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-15 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7668571
• 关键词: 3-Dimensional; Affect; Affinity; Amino Acid Substitution; Amino Acids; Antibodies; Antibody-mediated protection; Antigens; Avian Influenza; Avian Influenza A Virus; Avidity; Bacteriophages; Binding; Biological; Biological Assay; Canis familiaris; Cell Line; Cell surface; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; Charge; Chickens; Cleaved cell; Clinical Trials; Cloning; Complement; Computer Simulation; Containment; Crystallization; Data; Dissociation; Dose; E protein; Early treatment; Emerging Communicable Diseases; Ensure; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Epitopes; Equus caballus; Erythrocytes; Escape Mutant; Evaluation; Evolution; Future; Galactose; Genes; Genetic; Glycoproteins; Goals; H5 hemagglutinin; HIV-1; Harvest; Hemagglutination; Hemagglutinin; Human; Hybridomas; Image; Immune; Immunity; Immunization; Immunotherapy; In Vitro; Incubated; Individual; Infection; Influenza; Influenza A Virus, H5N1 Subtype; Kidney; Laboratories; Lentivirus Vector; Libraries; Link; Luciferases; MDCK cell; Maps; Measures; Mediating; Methods; Modeling; Molecular Virology; Monoclonal Antibodies; Mus; Mutagenesis; Mutate; Mutation; N-Acetylneuraminic Acid; Neuraminidase; New England; Onset of illness; Pan Genus; Passive Immunotherapy; Pathogenicity; Pathway interactions; Pattern; Peptides; Phage Display; Plasmids; Polymers; Polysaccharides; Population Heterogeneity; Prevention; Process; Property; Proteins; Recombinant Proteins; Recombinants; Reporter; Research; Research Infrastructure; Resolution; Ruthenium Ben; SARS coronavirus; Screening procedure; Serotyping; Severe Acute Respiratory Syndrome; Sialic Acids; Site; Specificity; Stochastic Processes; Structural Protein; Structure; Surface; Surface Plasmon Resonance; System; Techniques; Technology; Testing; Therapeutic; Time; United States; United States Dept. of Health and Human Services; Variant; Viral; Viral Load result; Viral Proteins; Virus; Virus Diseases; West Nile virus; Work; Yeasts; alpha-Fetoproteins; antibody engineering; base; biodefense; cost; density; design; egg; env Gene Products; experience; expression vector; fitness; gene cloning; glycosylation; high throughput screening; immunological diversity; in vitro activity; in vivo; influenzavirus; medical schools; member; microbial alkaline proteinase inhibitor; monolayer; mutant; neutralizing antibody; neutralizing monoclonal antibodies; novel; pandemic disease; pathogen; pressure; prevent; receptor; receptor binding; research study; response; restriction enzyme; sialic acid receptor; structural biology; tool; viral RNA

DESCRIPTION (provided by applicant): Highly pathogenic avian influenza (HPAI) virus H5N1 is an emerging infectious pathogen that is capable of causing a worldwide pandemic with widespread debilitating illness and death. There is an urgent need to understand MAPI H5N1 virus evolution and to develop therapeutic strategies to elicit protective host immunity, both passively and actively. In the case of HPAI H5N1, the importance of neutralizing antibodies in preventing disease onset is clearly established. This U01 Project represents the combined efforts of members of the Marasco (Antibody Engineering), Liddington (Structural Biology) and Donis (Molecular Virology) laboratories to investigate and further develop a novel anti-viral strategy that we term "Convergent Combination Immunotherapy (CCI)" for the prevention and early treatment of HPAI H5N1 virus infection. This approach involves the use of a cocktail of neutralizing human Mabs that is specifically formulated to have the broadest neutralization activity against emerging HPAI H5N1 variants. This novel concept is based on the understanding that influenza viruses undergo rapid neutralization escape under immune pressure. Our hypothesis, based on extensive preliminary data, is that it may be possible to focus sufficient immune pressure against critical neutralizing epitopes on hemagglutinin (H5) and neuraminidase (N1) HPAI to prevent neutralization escape or if it occurs, it will be at a great cost to viral fitness and pathogenicity. In this proposal, we will test 14 hypotheses that relate to defining the neutralizing epitopes on these two viral proteins and the effects of immune pressure on influenza virus evolution. In aim 1, we will isolate panels of high affinity human antibodies by phage display against all neutralizing epitopes on HAS and NA1 and will test their cross-neutralization activity using both H5/N1 pseudotype HIV-1 reporter viruses and clade 1/2 H5N1 strains. In aim 2 we will map the neutralization epitopes, determine antibody binding affinities and mechanisms of neutralization and perform co-crystallographic studies. In aim 3, we will perform extensive studies to examine pathways of neutralization escape. Central to our hypothesis, we will also test whether a cocktail of three neutralizing antibodies (CCI) to different neutralizing epitopes or sequential epitopes in a unique escape pathway can prevent neutralization escape. In aim 4 we will examine the cost of neutralization escape on viral fitness in vitro and will test the hypothesis that viruses that escape CCI are less pathogenic in vivo. We anticipate that 12-15 fully neutralizing human Mabs against HPAI H5N1 will be produced from our studies. The final goal of this proposal is to produce a cocktail of three Mabs, termed "TriAIVumab" that can be moved forward in to human clinical trials.

描述（由申请人提供）：高度致病的禽流感（HPAI）病毒H5N1是一种新兴的感染性病原体，能够引起全球大流行，并具有广泛的使人衰弱的疾病和死亡。迫切需要了解MAPI H5N1病毒的进化，并制定治疗策略，以被动和积极地引起保护性宿主的免疫力。就HPAI H5N1而言，清楚地确定了中和抗体在预防疾病发作中的重要性。该U01项目代表了Marasco（抗体工程），Liddington（结构生物学）和Donis（Molecular病毒学）实验室成员的综合努力，以调查和进一步制定一种新型的抗病毒病毒策略，我们称我们为预防和早期治疗Hpai Hpai Hpai H5N1 Virus Mettection“收敛组合联合治疗（CCI）”。这种方法涉及使用中和人mab的鸡尾酒，这些鸡尾酒的使用是针对新兴的HPAI H5N1变体具有最广泛的中和活性的。这个新颖的概念是基于这样的理解：在免疫压力下，流感病毒经历了快速中和逃逸。我们的假设基于广泛的初步数据，是，有可能将足够的免疫压力集中在血凝素（H5）和神经氨酸酶（N1）HPAI上，以防止中和逃脱，或者如果发生这种情况，它将为病毒适应性和病原体的基本付出很大的成本。在此提案中，我们将测试与定义这两种病毒蛋白的中和表位有关的14个假设以及免疫压力对流感病毒进化的影响。在AIM 1中，我们将通过噬菌体显示出对所有中和表位的HAS和NA1的噬菌体抗体隔离，并将使用H5/N1 Pseudotype HIV-1报告病毒和进化枝1/2 H5N1菌株测试其交叉中和中性化活性。在AIM 2中，我们将绘制中和表位，确定中和的抗体结合亲密关系和机制，并进行共结晶研究。在AIM 3中，我们将进行广泛的研究以检查中和逃生的途径。我们的假设的中心，我们还将测试三种中和抗体（CCI）的鸡尾酒是否在独特的逃生途径中与不同中和表位或顺序表位的鸡尾酒是否可以防止中和逃生。在AIM 4中，我们将检查体外病毒适应性的中和逃逸成本，并将检验以下假设：逃脱CCI的病毒在体内的病原体较低。我们预计将从我们的研究中产生12-15个完全中和对HPAI H5N1的人群。该提案的最终目标是生产三个单克隆抗体的鸡尾酒，称为“ triaivumab”，可以将其转移到人类的临床试验中。