RHINOVIRUS--HIV GP41 ELDKWA IMMUNOGENS FOR AIDS VACCINES

产品说明：（改编自申请人的摘要）能够引发针对HIV的原代分离物的中和免疫应答的HIV的少数区域之一是gp 41蛋白的外部结构域上的ELDKWA表位（以其氨基酸序列命名）。虽然并非所有感染者都对这一高度保守的区域产生免疫反应，但这些人更有可能具有良好的健康特征，并降低将HIV传播给后代的可能性，这突出了针对HIV这一区域进行疫苗接种的重要性。在这项工作中，我们建议构建组合库，系统地提出这个表位在一个自然的免疫原性表面环的冷引起的人鼻病毒的构象的多样性阵列。ELDKWA序列的侧翼为不同长度和序列的接头以及具有形成二硫键潜力的半胱氨酸残基。我们建议在这个库中产生大量潜在嵌合体的一小部分，以及四个库，它们是亲本库的特定子集，将编码类似于在罗格斯大学John Taylor教授的实验室中合成的一组平行的ELDKWA呈递约束肽的ELDKWA呈递。选择具有ELDKWA序列的免疫学相关表达的嵌合体，因为它们能够被中和人单克隆抗体2F 5识别。使用这些方法，我们已经能够引发一些针对HIV-1 V3环的有效中和免疫应答。将检测免疫选择病毒亚组与2F 5和HIV IG的反应性（使用ELISA和BIAcore实验），并检测嵌合病毒被2F 5和HIV IG中和的能力。最感兴趣的嵌合体将用于免疫豚鼠，以评估其针对不同的实验室和HIV-1原代分离株引发抗HIV-1中和应答的能力。亲和力和中和数据将用于指导第二代病毒文库（和肽）的设计，并在该资助的第二年进行表征。我们希望，这种设计ELDKWA免疫原性呈递的系统方法将导致鉴定能够引起对HIV的稳健、一致和交叉反应性免疫应答的免疫原。

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.