Do peptide mimetics of gp41 improve antibody-epitope interactions?

gp41 的肽模拟物是否可以改善抗体-表位相互作用？

基本信息

批准号：
8262547
负责人：
Vincent Joseph Venditto
金额：
$ 5.22万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-03-01 至 2015-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8262547
关键词：
Acquired Immunodeficiency Syndrome Address Affinity Alkanesulfonates Amino Acids Animals Antibodies Antibody Affinity Antibody Formation Antigens Aspartic Acid Autoimmune Process Binding Biological Assay Cells Chemicals Communicable Diseases Cysteic Acid Cysteine Data Development Drug Formulations Enzyme-Linked Immunosorbent Assay Epidemic Epitopes Evaluation Exhibits Extravasation Failure Freund&apos s Adjuvant Future Glutamic Acid HIV HIV Infections HIV vaccine HIV-1 Immune Sera Immune response Immunity Immunization Infection Inflammation Infusion procedures Irrigation Lead Lipid Bilayers Lipids Liposomes Mediating Membrane Modeling Modification Monoclonal Antibodies Mucosal Immune Responses Mucosal Immunity Mus Mutation Oryctolagus cuniculus Patients Peptides Phospholipids Phosphorylation Population Positioning Attribute Post-Translational Protein Processing Primates Proteins Reaction Sampling Serine/Threonine Phosphorylation Serum Site Structure-Activity Relationship Surface Testing Tyrosine Vaccinated Vaccines Viral Load result Virus Walkers Work abstracting analog base clinically relevant cross reactivity design env Gene Products immunogenic immunogenicity immunoreactivity improved in vitro Assay inorganic phosphate mimetics monophosphoryl lipid A neonatal Fc receptor neutralizing antibody neutralizing monoclonal antibodies nitration novel phosphonate polyclonal antibody prevent protein aminoacid sequence receptor binding success vaccine development

项目摘要

Do peptide mimetics of gp41 improve antibody-epitope interactions? Abstract In this F32 application I postulate that the broadly HIV neutralizing monoclonal antibodies, 2F5 and 4E10, against the membrane proximal external region (MPR) of gp41 were induced in a small population of donors by MPR epitopes due to modifications in the envelope proteins. My idea is to generate MPR immunogens incorporating unnatural amino acids that may induce stronger immune responses. I believe that these novel peptides will induce stronger affinity toward monoclonal antibodies, 2F5 and 4E10, and potentially induce higher titer polyclonal antibodies in a prime-boost immunization assay. The idea to use unnatural amino acids is unique. MPR immunogen design has focused on eliciting antibody responses that cross-react with phospholipids because the mAbs 2F5 and 4E10 exhibit unusual cross-reactivity with phospholipids. Incorporation of chemically modified amino acids into known epitopes may induce stronger immune responses. I will synthesize a group of MPR-based peptide sequences containing modified residues, which will be used to determine antibody affinity using a BIAcore assay. Structure activity relationships will be derived from these data and will guide subsequent rounds of chemically modified peptides to find sequences that will bind optimally to the monoclonal antibodies. I will then determine the immunogenicity of each peptide in rabbits using a prime-boost immunization strategy. Lipopeptides based on the peptides used in the BIAcore assay will be presented in a liposomal formulation to further enhance immunogenicity by presenting the epitopes in a lipid bilayer. Rabbits will be immunized subcutaneously and intramuscularly and antisera from these animals will be obtained and evaluated in a validated ELISA assay to determine the antibody titer induced by each peptide immunogen. Furthermore, formulations containing the same lipopeptides will be delivered intranasally to evaluate the mucosal immune response. Formulations will be prepared in the presence and absence of a peptide that binds and is transported by the neonatal Fc receptor (FcRn). ELISA assays will be used to determine the antibody responses in serum from SC/IM immunized animals, while mucosal lavage and serum samples will be evaluated from IN immunized animals. Differences in antisera titers from SC/IM immunized animals will be compared to IN immunized animals. Antisera and mucosal lavage samples with high titers in rabbits will be used in traditional inhibition assays to determine the neutralization activity of primary HIV-1 isolates in PBMCs. Success in generating neutralizing antibodies in rabbits will lead to future work to generate monoclonal antibodies. The chemically modified peptides that induce neutralizing antibodies in rabbits could provide a widely applicable epitope design strategy for the development of vaccines to a variety of infectious diseases including HIV.

GP41的肽模拟物是否改善抗体 - 质质相互作用？抽象的在此F32应用中，我假设，针对GP41的膜近端外部区域（MPR），在少数供体中，由于信封蛋白的修饰而引起的少数供体供体诱导了HIV中和HIV的单克隆抗体2f5和4e10。我的想法是生成掺入可能引起更强免疫反应的非天然氨基酸的MPR免疫原。我认为这些新型肽会引起对单克隆抗体2F5和4E10的更强亲和力，并可能在促进型免疫试验中诱导较高的滴度多克隆抗体。使用不自然氨基酸的想法是独一无二的。 MPR免疫原设计集中于引起与磷脂反应的抗体反应，因为MABS 2F5和4E10与磷脂表现出异常的交叉反应性。将化学改性的氨基酸掺入已知表位可能会引起更强的免疫反应。我将合成一组基于MPR的肽序列，这些肽序列含有修饰的残基，该序列将用于使用BIACORE分析来确定抗体亲和力。结构活性关系将从这些数据中得出，并将指导随后的化学修饰肽，以找到将最佳结合与单克隆抗体结合的序列。然后，我将使用最大促进免疫策略来确定兔子中每种肽的免疫原性。基于双胎测定中使用的肽的脂肽将以脂质体制剂呈现，以通过在脂质双层中呈现表位来进一步增强免疫原性。将在经过验证的ELISA测定中获得并评估兔子的皮下和肌内免疫，并从这些动物中进行抗血清，以确定每种肽免疫原诱导的抗体滴度。此外，将在鼻内递送包含相同脂蛋白肽的制剂以评估粘膜免疫反应。在存在和不存在结合并通过新生儿FC受体（FCRN）结合并运输的肽的情况下制定制剂。 ELISA分析将用于确定SC/IM免疫动物血清中的抗体反应，而粘膜灌洗和血清样品将在免疫动物中评估。将SC/IM免疫动物的抗血清滴度与免疫动物的差异进行比较。在传统的抑制测定中将使用抗血清和粘膜灌洗样品，以确定PBMC中原代HIV-1分离株的中和活性。在兔子中产生中和抗体的成功将导致将来的工作以产生单克隆抗体。诱导兔子中和抗体的化学修饰肽可以为将疫苗开发为包括HIV在内的各种传染病的开发提供广泛适用的表位设计策略。