Defining clinically relevant viral epitopes with cow antibodies

用牛抗体定义临床相关的病毒表位

• 批准号: 10014637
• 负责人: Vaughn Vasil Smider
• 金额: $ 35.86万
• 依托单位: APPLIED BIOMEDICAL SCIENCE INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10014637
• 关键词: Amino Acids; Animal Model; Animals; Antibodies; Antibody Repertoire; Antibody Response; Antigens; Binding; Biochemical; Cattle; Common Epitope; Communicable Diseases; Cysteine; Data; Development; Electron Microscopy; Engineering; Epitopes; Flavivirus; Future; Genetic Engineering; HIV; HIV Antibodies; HIV Envelope Protein gp120; Health; Human; Immune response; Immunization; Immunize; Immunoglobulins; Immunology; Lead; Length; Maps; Medical; Molecular; Monoclonal Antibodies; Mus; Mutagenesis; Mutation; Outcome; Outcome Study; Polysaccharides; Positioning Attribute; Property; Protein Engineering; Proteins; Retroviridae; Structure; Structure-Activity Relationship; Surface; System; Techniques; Texas; Trastuzumab; Vaccination; Vaccine Design; Vaccines; Viral; Virus; Virus Diseases; X-Ray Crystallography; antigen antibody binding; antigen binding; clinically relevant; deep sequencing; disulfide bond; experimental study; humanized antibody; insight; knob protein; neutralizing antibody; neutralizing monoclonal antibodies; next generation; novel; novel vaccines; prevent; structural biology; success; tool; vaccine development; virology; ward

Vaccines are the primary means by which to prevent, control, or eradicate infectious diseases. While many vaccines have been successfully developed and have resulted in enormous medical and veterinary benefit, there are certain viruses that have eluded effective vaccine development. Generally, viruses with multiple strains or that have high mutation rates can evade neutralizing antibodies, as their surface determinants are variable and result in the inability of neutralizing antibodies raised against one strain to bind and neutralize alternative strains. Certain rare epitopes, however, are required for viral infection and are conserved across strains. Interestingly, neutralizing antibodies against these rare epitopes tend to have long CDR H3 regions. In the case of HIV, long CDR H3s can pierce the viral glycan shield and reach into the conserved epitope on the gp120 spike protein. While long CDR H3 regions in human antibodies are infrequent in the repertoire, cattle routinely produce long (20-40 amino acids) and ultralong (40-70 amino acids) CDR H3 regions that have unique “stalk” and “knob” structural features that protrude far from the antibody surface. Therefore, cattle may be an excellent model organism to identify and define new and conserved neutralizing epitopes in these challenging viruses. Indeed, in preliminary experiments we have found that cattle make a robust and broadly neutralizing antibody response to the HIV gp120 antigen. Here we propose to use the unique cow antibody repertoire to define new conserved neutralizing epitopes on two viruses of great importance to human and animal health, HIV and BVDV. Effective vaccines against both of these viruses have been a major challenge to develop. We will immunize animals against these viruses, generate monoclonal antibodies that neutralize the virus as well as related strains, and molecularly map the antigen-antibody interaction using mutagenesis and structural biology techniques. Definition of new conserved epitopes could lead to engineered epitope-specific vaccines. Thus, the outcomes of this proposal could enable generation of next-generation vaccines for these two viruses, but could also have broad utility in vaccine development against other challenging viruses in the future.

疫苗是预防、控制或根除传染性疾病的主要手段。 疾病虽然许多疫苗已经成功开发并导致了 巨大的医疗和兽医效益，有一些病毒已经逃脱 有效的疫苗开发。通常，具有多个毒株或具有高 突变率可以逃避中和抗体，因为它们的表面决定簇是 可变的，并导致针对一种菌株产生的中和抗体不能 结合并中和替代菌株。然而，某些罕见的表位是必需的， 病毒感染，并且在菌株间是保守的。有趣的是，中和抗体 针对这些罕见表位的抗体倾向于具有长CDR H3区。在艾滋病方面，长期 CDR H3可以穿透病毒聚糖屏蔽并到达病毒上的保守表位。 gp 120刺突蛋白。虽然人抗体中的长CDR H3区在免疫组织化学中是罕见的， 牛通常产生长（20-40个氨基酸）和超长（40-70个氨基酸 氨基酸）CDR H3区具有独特的“茎”和“节”结构特征， 远离抗体表面突出。因此，牛可能是一个极好的模型 生物体中鉴定和定义新的和保守的中和表位， 挑战病毒事实上，在初步的实验中，我们发现牛 对HIV gp 120抗原的稳健和广泛中和抗体应答。这里我们 建议使用独特的牛抗体库来定义新的保守中和抗体， 对人类和动物健康非常重要的两种病毒，HIV和 BVDV。针对这两种病毒的有效疫苗一直是一个重大挑战， 开发.我们将使动物免疫这些病毒， 中和病毒以及相关菌株的抗体，并对 使用诱变和结构生物学技术的抗原-抗体相互作用。 新的保守表位的定义可能导致工程化的表位特异性疫苗。 因此，该提案的结果可以使下一代 这两种病毒的疫苗，但也可能在疫苗开发中具有广泛的用途 对抗其他具有挑战性的病毒