Engineering Simian-derived Lactobacilli to Secrete Anti-HIV-1 Microbicides

改造猴源性乳酸杆菌以分泌抗 HIV-1 杀菌剂

• 批准号: 7174370
• 负责人: DANIEL BODEN
• 金额: $ 21.49万
• 依托单位: AARON DIAMOND AIDS RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-15 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7174370
• 关键词: Macaca; antiinfective agents

DESCRIPTION (provided by applicant): Every day nearly 14,000 people are newly infected with HIV-1 and over 80 % of these infections are transmitted by the mucosal route during unprotected rectal or vaginal intercourse. The development of a safe and effective topical microbicide may offer a preventative strategy that could have a major impact on the course of the HIV pandemic. Six candidate topical microbicides are currently undergoing clinical effectiveness trials and many others are under development. However, potential shortcomings of topical microbicides are: the requirement of immediate pre-coital application, possible vaginal irritation/inflammation upon long-term use which may result in enhanced HIV-1 transmission, and relatively high production costs. The strategy of using live microbial microbicides may solve the potential problems inherent to chemically based microbicides. Here, we seek to develop a live microbial anti-HIV microbicide based on simian-derived vaginal Lactobacilli isolates. The cervicovaginal mucosa is primarily colonized by commensal Lactobacilli and the secretion of substantial quantities of an anti-HIV microbicide by genetically engineered Lactobacilli may effectively block vaginal transmission of HIV-1. We will identify vaginal Lactobacilli isolates from macaques which demonstrate high hydrogen peroxide (H2O2) production and strong adherence to vaginal epithelial cells, critical factors for in situ colonization. Plasmid systems will be engineered to express and secrete anti-HIV compounds. The ability of recombinant Lactobacilli strains to colonize the cervicovaginal tract will be tested in rhesus macaques. Finally, macaques will be inoculated with recombinant Lactobacillus secreting anti-HIV microbicides and subsequently challenged with chimeric/simian immunodeficiency virus SHIV-162P3 to determine protective efficacy. The goal of this application is to develop a live microbial topical microbicide against HIV-1 based on vaginal Lactobacillus isolates that may offer the following significant advantages over chemically based topical microbicides: 1) Long-lasting protection due to bacterial colonization, thus eliminating the need for direct pre-coital application and more control by the vulnerable receptive partner; 2) Low potential to induce inflammatory reactions by using vaginal commensal bacteria; and 3) Low production costs compared to protein-based microbicides.

描述（由申请人提供）：每天有近14,000人新感染HIV-1，其中80％以上的感染在未受保护的直肠或阴道性交期间通过粘膜途径传播。安全有效的局部杀菌剂的发展可能会提供一种预防策略，可能会对HIV大流行的进程产生重大影响。目前正在进行六个候选局部杀菌剂，正在进行临床有效性试验。但是，局部菌皮剂的潜在缺点是：立即使用前应用前应用，长期使用后可能的阴道刺激/炎症，这可能会导致HIV-1传播增强，以及相对较高的生产成本。使用实时微生物微生物的策略可以解决化学基菌物固有的潜在问题。在这里，我们寻求基于邻氨酸衍生的阴道乳酸乳杆菌分离株开发活微生物抗HIV微生物。子宫颈阴道粘膜主要是通过共生乳酸杆菌定植的，并且通过基因设计的乳酸杆菌分泌大量大量的抗HIV微生物剂，可能有效地阻止了HIV-1的阴道传播。我们将从猕猴中鉴定出阴道乳酸杆菌分离株，这些乳酸是表现出高过氧化氢（H2O2）的高氢（H2O2）的产生，并且对阴道上皮细胞的依从性很强，这是原位定植的关键因素。质粒系统将被设计为表达和分泌抗HIV化合物。重组乳酸菌菌株定居宫颈阴道区的能力将在恒河猕猴中进行测试。最后，猕猴将被分泌抗HIV杀菌剂的重组乳酸杆菌接种，随后用嵌合/邻近的邻苯二醇免疫缺陷病毒SHIV-162P3挑战以确定保护性。该应用的目的是基于阴道乳酸杆菌分离株开发针对HIV-1的实用微生物局部杀菌剂，该分离株可能具有以下重要优势，而不是化学基于化学的局部微生物剂：1）由于细菌定殖而导致的持久保护，从而消除了对直接的预科应用的需求，并消除了易于受到易于接受的接收伙伴的控制。 2）使用阴道共生细菌诱导炎症反应的潜力低； 3）与基于蛋白质的微生物相比，生产成本低。