A SCALABLE PHOTOCHEMICAL INACTIVATION PROCESS FOR HIV

可扩展的 HIV 光化学灭活工艺

• 批准号: 6374536
• 负责人: HAYNES W SHEPPARD
• 金额: $ 16.2万
• 依托单位: PUBLIC HEALTH FOUNDATION ENTERPRISES
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6374536
• 关键词: AIDS vaccines; Animalia; HIV envelope protein gp120; active immunization; mass tissue /cell culture; photochemistry; psoralens; ultraviolet radiation; vaccine development

A traditionally successful approach to viral vaccines, whole inactivated virus (WIV), has received relatively little attention in the overall effort to develop an effective HIV/AIDS vaccine. Only one inactivated virus product (the Salk/IRC Immunogen) has been tested in humans and this product is envelope depleted and has been tested only as a therapeutic rather than a prophylactic immunogen. Since major target epitopes for antibody neutralization are in the envelope glycoprotein, it is generally considered a desirable component of a prophylactic immunogen. Thus far, however, immunization of humans with monomeric recombinant envelope glycoprotein has resulted in relatively type-specific neutralization and very little neutralization of primary isolates. The few monoclonal antibodies that demonstrate broad primary- isolate neutralization generally recognize conformation dependent epitopes. Such epitopes might be retained on whole inactivated virions. Photochemical treatment (PCT) with psoralen and UV light is a promising approach to inactivating HIV without disrupting "native" oligomeric structures. We propose to investigate a potentially scaleable process using a proprietary variant of psoralen (5-59) that lacks the in vitro mutagenicity associated with conventional psoralen methods. In addition, 5-59 inactivated products have undergone extensive safety testing in humans. The objectives of this study will be l) to investigate methods to produce and concentrate pilot-scale preparations of HIV with minimal envelope loss, 2) to confirm and Optimize a photochemical treatment process, which will use the novel psoralen 5-59, and UVA to inactivate HIV to an acceptable level while retaining gpl20 and its conformational epitopes, 3) to design/construct a novel flow PCT that is capable of treating laboratory scale quantities (5 L) of HIV culture and is capable of being scaled-up to treat production scale quantities (5000 L), 4) investigate and identify potential process steps that can be used to reduce levels of residual psoralen while maintaining native gp120 structure, 5) investigate potential interactions between PCT and other inactivation processes and determine an optimal process sequence and conditions for obtaining a double- inactivated WIV with intact gpl20, and 6) investigate animal immunogenicity of these preparations.

在开发有效的艾滋病毒/艾滋病疫苗的总体努力中，传统上成功的病毒疫苗方法--全灭活病毒(WIV)--相对较少受到关注。只有一种灭活病毒产品(Salk/IRC免疫原)在人类身上进行了测试，该产品的包膜被耗尽，仅作为治疗性免疫原进行了测试，而不是作为预防免疫原进行测试。由于抗体中和的主要靶位位于包膜糖蛋白中，因此它通常被认为是预防性免疫原的理想成分。然而，到目前为止，用单体重组包膜糖蛋白免疫人类，导致了相对特定类型的中和，而初级分离株的中和很少。少数表现出广泛的初级分离中和作用的单抗一般识别构象依赖的表位。这样的表位可能保留在整个灭活的病毒粒子上。补骨脂素和紫外光的光化学治疗(PCT)是一种很有前途的方法，可以在不破坏“天然”低聚结构的情况下灭活HIV。我们建议使用补骨脂素的专有变体(5-59)来研究一种潜在的可扩展的过程，该方法缺乏与传统补骨脂素方法相关的体外诱变性。此外，5-59种灭活产品已经在人体上进行了广泛的安全测试。本研究的目的将是：(L)研究以最小的包膜损失生产和浓缩艾滋病毒的中试制剂的方法；2)确认和优化光化学处理工艺，其将使用新型补骨脂素5-59和UVA将艾滋病毒灭活到可接受的水平，同时保留gpl20及其构象表位；3)设计/构造能够处理实验室规模的艾滋病毒培养物(5 L)并且能够被放大以处理生产规模数量(5000 L)的新型流动PCT；4)调查和确定可用于在保持本地gp120结构的同时降低补骨脂素残留水平的潜在工艺步骤，5)研究PCT与其他灭活过程之间的潜在相互作用，确定获得完整gpl20双灭活WIV的最佳工艺顺序和条件，以及6)考察这些制剂的动物免疫原性。