Project 1: Immunogens and Manufacturing

项目一：免疫原及制造

基本信息

批准号：
8478317
负责人：
HARRIET L ROBINSON
金额：
$ 185.9万
依托单位：
GEOVAX, INC.
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8478317
关键词：
3&apos Untranslated Regions AIDS/HIV problem Address Adjuvant Animals Antibodies Antibody Formation Antigens Avidity B-Lymphocytes Binding Bioreactors Birds Calmodulin Cell Line Cells Chick Embryo Clinical Research Collaborations Country Developed Countries Development Ducks Fibroblasts Frameshift Mutation France GMP lots Generations Genes Goals Growth HIV HIV Vaccine Trials Network HIV vaccine Human Individual Interruption Laboratories Length Life Macaca mulatta Mammalian Cell Membrane Methods MicroRNAs Modeling Mosses National Institute of Allergy and Infectious Disease Pharmaceutical Preparations Phase Point Mutation Positioning Attribute Preparation Process Production Recombinants Running SIV Seeds Silent Mutation Stem cells TNFSF5 gene Testing Time Toxic effect Treatment Protocols Vaccination Vaccine Design Vaccine Production Vaccines Virus Virus Diseases Virus-like particle Work cell bank continuous cell line design drug resistant virus gp160 improved manufacturing process meetings mutant prevent programs research clinical testing response safety testing stability testing stem vector

项目摘要

Our goal is to design and produce a clade C HIV MVA vaccine that will elicit antibodies capable of preventing HIV acquisition. Our clade B HIV VLP-expressing MVA-vectored vaccine has been tested clinically and shown good ability to Induce Env-specific antibodies. The clade B HIV vaccine, however, is not designed to express full length Env that we have found to prime the most protective antibody responses using the SIV rhesus macaque model (see Program Overview). Two technical approaches are proposed to increase the induction of protective Ab responses; the development of a MVA-vectored HIV vaccine encoding full-length, clade 0 gp160 and the use of MVA-expressed CD40L as a B-cell adjuvant. Specifically,we will 1) Construct a clade C VLP MVA vaccine expressing full-lengfh gp160 Env to optimize the protective avidity and neutralizing activity of elicited Env-specific Ab, 2) Insert target sequences for EB66 stem cellspecific miRNAs into 3' untranslated regions of vaccine Env inserts to suppress insert expression and enhance titers and vector stability during manufacture in the EB66 cell line and 3) Construct CD40Lexpressing MVA to serve as a strong B cell adjuvant for the titer and avidity of Env-specific Ab. Production of MVA vaccines is a challenge because of the growth of these vaccines in chick embryo fibroblasts that require sourcing and producing large batches of primary cells. Because of this, GeoVax has explored the use of continuous cell lines for growth of MVA and has found the EB66 duck stem cell-derived line developed by Vivalis LLC. Nantes France to be a promising option. GeoVax has produced dedicated Master and Working EB66 cell banks. The goals for this project include 1) Qualification of media and production processes for EB66 cells as the MVA manufacturing cell substrate focusing on "end-of-production" testing and the generation of Master seed stocks, 2) Production of a "GMP-like" pilot/feasibility lot of MVA-vectored clade C vaccine and the MVA encoding CD40L to validate the manufacturing process and to produce material for animal safety testing and 3) Production of a GMP lot of each vaccine to be used in Phase 1 clinical testing by the HVTN.

我们的目标是设计和生产 C 分支 HIV MVA 疫苗，该疫苗将引发能够产生以下抗体的抗体：预防艾滋病毒感染。我们的 B 进化枝 HIV VLP 表达 MVA 载体疫苗已经过测试临床上显示出良好的诱导环境特异性抗体的能力。然而，进化枝 B HIV 疫苗并非旨在表达全长 Env，我们发现全长 Env 可以使用 SIV 恒河猴模型引发最具保护性的抗体反应（参见项目概述）。提出了两种技术方法来增强保护性抗体反应的诱导；开发编码全长进化枝 0 gp160 的 MVA 载体 HIV 疫苗，并使用 MVA 表达的 CD40L 作为 B 细胞佐剂。具体来说，我们将 1) 构建表达全长 gp160 Env 的进化枝 C VLP MVA 疫苗，以优化引发的 Env 特异性抗体的保护亲合力和中和活性，2) 将 EB66 干细胞特异性 miRNA 的靶序列插入到 EB66 干细胞特异性 miRNA 的 3' 非翻译区中。疫苗 Env 插入物可抑制插入物表达并增强 EB66 细胞系生产过程中的滴度和载体稳定性，以及 3) 构建表达 CD40L 的 MVA 作为强 B 细胞佐剂，可提高 Env 特异性抗体的效价和亲合力。 MVA 疫苗的生产是一项挑战，因为这些疫苗是在鸡胚中生长的需要采购和生产大批量原代细胞的成纤维细胞。因此，GeoVax 探索了使用连续细胞系来生长 MVA，并发现了由 Vivalis LLC 开发的 EB66 鸭干细胞衍生系。法国南特是一个有前途的选择。 GeoVax 生产了专用的主细胞库和工作 EB66 细胞库。该项目的目标包括 1) EB66 细胞作为 MVA 制造细胞基质的培养基和生产工艺的资格，重点关注“生产结束”测试和主种子库存的生成，2) 生产“类 GMP” “ MVA 载体 C 分支疫苗和编码 CD40L 的 MVA 的试点/可行性批次，以验证生产过程并生产用于动物安全测试的材料，以及 3) 生产将在阶段中使用的每种疫苗的 GMP 批次1 由 HVTN 进行临床测试。