RHINOVIRUS--HIV GP41 ELDKWA IMMUNOGENS FOR AIDS VACCINES

描述：（改编自申请人摘要）在gp41蛋白的外部结构域上，ELDKWA表位（以其氨基酸序列命名）是少数几个能够引发针对HIV初级分离株的中和性免疫反应的HIV区域之一。虽然并非所有感染者都对这一高度保守的区域有免疫反应，但那些对这一区域有免疫反应的人更有可能具有良好的健康特征，并且将艾滋病毒传播给后代的可能性降低，这突出了针对这一艾滋病毒区域进行疫苗接种的重要性。在这项工作中，我们建议构建组合文库，系统地在引起感冒的人鼻病毒的自然免疫原性表面环上以多种构象呈现该表位。ELDKWA序列的两侧是不同长度和序列的连接体，以及具有形成二硫键的潜力的半胱氨酸残基。我们打算利用这个库中大量潜在嵌合体的一小部分，以及四个库，这些库是母库的特定子集，它们将编码ELDKWA表达，类似于Rutgers大学John Taylor教授实验室合成的ELDKWA表达约束肽的平行集。将选择具有ELDKWA序列免疫学相关表现的嵌合体，因为它们能够被中和的人单克隆抗体2F5识别。使用这些方法，我们已经能够引发一些有效的中和免疫反应，报道针对HIV-1 V3环。免疫选择的一部分病毒将被测试与2F5和HIV IG的反应性（使用elisa和BIAcore实验），嵌合病毒也将被测试它们被2F5和HIV IG中和的能力。最令人感兴趣的嵌合体将用于免疫豚鼠，以评估它们对各种实验室和原代HIV-1分离株引发抗HIV-1中和反应的能力。亲和性和中和性数据将用于指导第二代病毒库（和肽）的设计，这些病毒库将在本资助的第二年制作和表征。我们希望这种设计ELDKWA免疫原性表达的系统方法将导致免疫原的鉴定，这些免疫原将能够引发对HIV的强大，一致和交叉反应性免疫反应。

DESCRIPTION: (Adapted from Applicant's Abstract) One of the few regions of HIV capable of eliciting neutralizing immune responses against primary isolates of HIV is the ELDKWA epitope (named for its amino acid sequence) on the exterior domain of the gp41 protein. While not all infected individuals respond immunologically to this highly conserved region, those who do are more likely to have favorable health characteristics and a reduced likelihood of transmitting HIV to offspring, highlighting the importance of targeting this region of HIV for vaccine purposes. In this work, we propose to construct combinatorial libraries that systematically present this epitope in a diverse array of conformations on a naturally immunogenic surface loop of the cold causing human rhinovirus. The ELDKWA sequence will be flanked by linkers of varying lengths and sequences as well as by cysteine residues with the potential to form disulfide bonds. We propose to make a fraction of the enormous number of potential chimeras in this library as well as four libraries that are specific subsets of the parent library that will encode ELDKWA presentations analogous to those of a parallel set of ELDKWA presenting constrained peptides being synthesized in the laboratory of Professor John Taylor at Rutgers University. Chimeras with immunologically relevant presentations of the ELDKWA sequence will be selected for their abilities to be recognized by the neutralizing human monoclonal antibody 2F5. Using these methods, we have been able to elicit some of the potent neutralizing immune responses reported against the HIV-1 V3 loop. A subset of the immunoselected viruses will be tested for reactivity with 2F5 and HIV IG (using ELISAs and BIAcore experiments) and chimeric viruses will be tested for their ability to be neutralized by 2F5 and HIV IG as well. Chimeras of greatest interest will be used to immunize guinea pigs to assess their ability to elicit anti-HIV-1 neutralizing responses against diverse laboratory and primary isolates of HIV-1. The affinity and neutralization data will be used to guide the design of second generation virus libraries (and peptides) to be made and characterized in the second year of this grant. It is our hope that such a systematic approach to the design of immunogenic presentations of ELDKWA will lead to the identification of immunogens that will be able to elicit robust, consistent, and cross-reactive immune responses to HIV.