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: 7287551
• 负责人: Wayne A. Marasco
• 金额: $ 159.44万
• 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 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7287551
• 关键词: 3-Dimensional; Affect; Affinity; Amino Acid Substitution; Amino Acids; Antibodies; Antibody-mediated protection; Antigens; Appendix; 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; Class; Cleaved cell; Clinical Trials; Cloning; Complement; Computer Simulation; Condition; Containment; Crystallization; Data; Dissociation; Doctor of Medicine; Doctor of Philosophy; 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; Human Parainfluenza Virus 1; 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; Numbers; Onset of illness; Pan Genus; Passive Immunotherapy; Pathogenicity; Pathway interactions; Pattern; Peptides; Phage Display; Plasmids; Polymers; Polysaccharides; Population Heterogeneity; Prevention; Principal Investigator; Process; Property; Proteins; Range; Rate; Recombinant Proteins; Recombinants; Reporter; Research; Research Infrastructure; Research Personnel; 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; concept; cost; day; density; design; egg; env Gene Products; experience; expression vector; fitness; gene cloning; glycosylation; high throughput screening; immunological diversity; in vivo; influenzavirus; medical schools; member; microbial alkaline proteinase inhibitor; monolayer; mutant; neutralizing antibody; neutralizing monoclonal antibodies; novel; pandemic disease; pathogen; pressure; prevent; programs; 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）：高致病性禽流感（HPAI）病毒 H5N1 是一种新出现的传染性病原体，能够引起世界范围内的大流行，导致广泛的衰弱性疾病和死亡。迫切需要了解 MAPI H5N1 病毒的进化，并制定治疗策略以被动和主动地引发保护性宿主免疫。就 HPAI H5N1 而言，中和抗体在预防疾病发作中的重要性已得到明确证实。这个 U01 项目代表了 Marasco（抗体工程）、Liddington（结构生物学）和 Donis（分子病毒学）实验室成员的共同努力，以调查和进一步开发一种新的抗病毒策略，我们称之为“聚合组合免疫疗法（CCI）” “用于预防和早期治疗 HPAI H5N1 病毒感染。该方法涉及使用中和人单克隆抗体混合物，该混合物经过专门配制，对新出现的 HPAI H5N1 变种具有最广泛的中和活性。这一新颖的概念基于流感病毒在免疫压力下经历快速中和逃逸的理解。基于广泛的初步数据，我们的假设是，有可能针对血凝素 (H5) 和神经氨酸酶 (N1) HPAI 上的关键中和表位集中足够的免疫压力，以防止中和逃逸，否则如果发生这种情况，后果将非常严重。病毒适应性和致病性的成本。在本提案中，我们将测试 14 个假设，这些假设涉及定义这两种病毒蛋白上的中和表位以及免疫压力对流感病毒进化的影响。在目标 1 中，我们将通过针对 HAS 和 NA1 上所有中和表位的噬菌体展示分离高亲和力人类抗体组，并将使用 H5/N1 假型 HIV-1 报告病毒和进化枝 1/2 H5N1 毒株测试它们的交叉中和活性。在目标 2 中，我们将绘制中和表位图，确定抗体结合亲和力和中和机制，并进行共晶体研究。在目标 3 中，我们将进行广泛的研究来检查中和逃逸的途径。我们的假设的核心是，我们还将测试针对不同中和表位或独特逃逸途径中的连续表位的三种中和抗体（CCI）的混合物是否可以防止中和逃逸。在目标 4 中，我们将检查中和逃逸对体外病毒适应性的成本，并将检验逃逸 CCI 的病毒在体内致病性较低的假设。我们预计我们的研究将产生 12-15 种针对 HPAI H5N1 的完全中和人类单克隆抗体。该提案的最终目标是生产三种单克隆抗体的混合物，称为“TriAIVumab”，可以进入人体临床试验。