Development of Novel Immunogens for Vaccines to HIV-1

HIV-1 疫苗新型免疫原的开发

• 批准号: 7615556
• 负责人: Robert G. Whalen
• 金额: $ 78.13万
• 依托单位: MAXYGEN, INC.
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7615556
• 关键词: Adopted; Amino Acids; Antigens; Binding; Collaborations; Complex; Consensus; Crystallization; Development; Directed Molecular Evolution; Epitopes; GP 140; Genetic Recombination; Goals; HIV; HIV Antigens; HIV-1; Immune response; Immunization; In Vitro; Methods; Molecular Conformation; Monoclonal Antibodies; Numbers; Oryctolagus cuniculus; Property; Proteins; Screening procedure; Structure; Surface; Technology; Testing; Vaccines; Variant; Viral; Virus; Virus-like particle; chimeric gene; clinically relevant; env Gene Products; glycosylation; immunogenic; immunogenicity; neutralizing antibody; neutralizing monoclonal antibodies; novel; programs

The main objective of this Proposal is to identify one or more novel HIV-1 envelope (Env) proteins capable of inducing broadly neutralizing antibodies to numerous clinically relevant primary viral strains. This Proposal employs a powerful technology called directed molecular evolution. This approach involves the use of in vitro recombination to create a large number of chimeric genes encoding variant Env structures that will be systematically evaluated with respect to their antigenic and immunogenic properties. Our hypothesis is that some of these novel variant proteins will adopt conformations in which conserved neutralizing epitopes will induce broadly cross-reactive neutralizing antibodies. Two principal screening approaches are (i) the use of monoclonal antibodies that broadly neutralize HIV-1 and (ii) immunization studies in rabbits. In vitro recombination and the associated screening methods will first be applied to glycosylation variants of the gp140 Env protein. In a second step, amino acid diversity will be incorporated into the best glycosylation variant using consensus HIV sequences from Group M viruses. In a third step, directed molecular evolution will be used in a recursive manner to achieve further improvement in the immunogenicity of these Env variants. The best immunogens will also be tested in a soluble trimeric form by Project 2. Novel Env antigens that strongly bind the existing monoclonal antibodies that neutralize HIV-1 will be prepared for boosting the neutralizing immune response. In collaboration with Project 2, we will used directed molecular evolution to assist in the crystallization of the Env molecule and to select for virus-like particles that carry stable Env complexes on their surface. This Project will contribute to the overall goals of the HIVRAD Program in developing approaches to increase the immunogenicity of HIV antigens.

该提案的主要目的是鉴定一种或多种新型HIV-1包膜（ENV）蛋白，能够诱导大量与许多临床相关的原发性病毒菌株诱导广泛中和抗体。该提案采用了一种强大的技术，称为指示分子进化。这种方法涉及使用体外重组来创建大量编码变体环境结构的嵌合基因，这些基因将对它们的抗原和免疫原性进行系统地评估。我们的假设是，其中一些新型的变体蛋白将采用构象，其中保守的中和表位将引起广泛的交叉反应性中和抗体。两种主要筛查方法是（i）使用兔子中的HIV-1和（ii）免疫研究的单克隆抗体的使用。 体外重组和相关的筛选方法将首先应用于GP140 ENV蛋白的糖基化变体。在第二步中，使用来自组M病毒的共识HIV序列将氨基酸多样性纳入最佳糖基化变体中。在第三步中，将以递归方式使用定向的分子进化，以进一步改善这些ENV变体的免疫原性。最好的免疫原子还将通过项目2以可溶性三聚体进行测试。新颖的ENV抗原强烈结合现有的单克隆抗体，中和HIV-1的现有单克隆抗体将为增强中和中和免疫反应做好准备。通过与项目2合作，我们将使用定向的分子进化来协助ENV分子的结晶，并选择在其表面上携带稳定Env复合物的病毒样颗粒。该项目将有助于Hivrad的整体目标 开发方法以增加HIV抗原的免疫原性的方法。