MINI-HIV VARIANTS AS LIVE-ATTENUATED VACCINE STRAIN

描述：(改编自申请者摘要)我们提出了一个新的方法 朝着构建安全、基因稳定的下一代艾滋病毒-1迈进 作为减毒活疫苗的变种。当代HIV-1 缺失变体不会永久减弱，因此是不安全的。我们会 用优化的组织培养进化系统逐步迫使HIV-1 从一种高效复制的复杂逆转录病毒进化而来，这种逆转录病毒编码九种 一种具有三到五个基因的简单病毒的蛋白质仍然能够 高效复制。假设逆转录病毒的自然进化开始 对于获得额外基因功能的更简单的形式，我们建议 将进化方向转向更简单的HIV-1变种。 例如，在试点进化实验中，我们成功地选择了 快速复制的HIV-1变种，缺乏三个辅助基因。数列 分析和重建实验应该揭示出 策略。这些快速复制的变体将 用于另一轮基因缺失和随后的进化以恢复 复制容量。因此，基因缺失和进化的重复循环 提出了适应，目的是获得一种复制的HIV-1变异株 最小数量的基因。迷你HIV作为疫苗的潜在用途 应变最初是由霍华德·特明提出的，尽管他建议 产生这种试剂的途径完全不同。额外的安全性 这些治疗病毒的特征将被纳入这些治疗病毒中，我们还将 削弱病毒基因的基本集合。这些结合起来的方法应该 产生一种安全的迷你HIV变种，可用作减毒活疫苗 紧张。这些病毒将等待广泛的复制测试以验证其 遗传稳定性，然后进行动物试验以筛选其致病因素 以及它们诱导保护性免疫反应的能力。

DESCRIPTION: (Adapted from Applicant's Abstract) We propose a novel approach towards the construction of a next generation of safe, genetically stable HIV-1 variants as live-attenuated AIDS vaccines. The current generation of HIV-1 deletion variants is not permanently attenuated, and therefore unsafe. We will use an optimized tissue culture evolution system to gradually force HIV-1 to evolve from an efficiently replicating, complex retrovirus that encodes nine proteins to a simple virus with three-to-five genes that is still able to replicate efficiently. Assuming that natural evolution of retroviruses started with the more simple forms that acquired additional gene functions, we propose to turn around the direction of evolution towards more simple HIV-1 variants. For instance, in pilot evolution experiments, we succeeded in selecting fast-replicating HIV-1 variants that lack three accessory genes. Sequence analysis and reconstruction experiments should reveal the "compensatory strategies" used by the revertant viruses. These fast-replicating variants will be used for another round of gene deletion and subsequent evolution to regain replication capacity. Thus, repeated cycles of gene deletion and evolutionary adaptation are proposed with the aim to obtain a replicating HIV-1 variant with a minimal number of genes. The potential use of mini-HIV versions as vaccine strain was proposed originally by Howard Temin, although he suggested a completely different route to generate such reagents. Additional safety features will be incorporated into these therapeutic viruses, and we will also attenuate the basic set of viral genes. These combined approaches should generate a safe mini-HIV variant that can be used as a live-attenuated vaccine strain. These viruses will await extensive replication tests to verify their genetic stability, followed by animal tests to screen for their pathogenic potential and their ability to induce a protective immune response.