RHINOVIRUS--HIV GP41 ELDKWA IMMUNOGENS FOR AIDS VACCINES

描述：(根据申请人的摘要改编)HIV的少数几个区域之一能够对HIV的主要分离株产生中和免疫反应，其中一个区域是gp41蛋白外部区域上的ELDKWA表位(因其氨基酸序列而命名)。虽然并不是所有的感染者对这个高度保守的区域都有免疫反应，但那些对这种高度保守的区域有免疫反应的人更有可能具有良好的健康特征，并降低将艾滋病毒传播给后代的可能性，这突显了针对这一区域的艾滋病毒用于疫苗目的的重要性。在这项工作中，我们建议构建组合文库，在引起人鼻病毒感冒的自然免疫原性表面环上以不同的构象系统地呈现该表位。ELDKWA序列的两侧将有不同长度和序列的连接子以及可能形成二硫键的半胱氨酸残基。我们建议将该文库中巨大数量的潜在嵌合体以及四个文库中的一小部分作为父库的特定子集，这些文库将编码ELDKWA演示文稿，类似于罗格斯大学John Taylor教授的实验室中合成的一组平行的ELDKWA演示文库。具有免疫相关的ELDKWA序列的嵌合体将被选择，因为它们的能力可以被中和的人类单抗2F5识别。使用这些方法，我们已经能够诱导针对HIV-1V3环路的一些有效的中和免疫反应。免疫选择病毒的一个子集将被测试与2F5和HIV IG的反应性(使用ELISA和Biacore实验)，嵌合病毒将被2F5和HIV IG中和的能力测试。人们最感兴趣的嵌合体将被用于免疫豚鼠，以评估它们对不同的HIV-1实验室和初级分离株产生抗HIV-1中和反应的能力。亲和力和中和数据将用于指导第二代病毒库(和多肽)的设计，这些库(和多肽)将在这笔赠款的第二年制作和表征。我们希望，这种设计ELDKWA免疫原性陈述的系统方法将导致识别能够引起对艾滋病毒的强大、一致和交叉反应的免疫原。

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.