Genetic Scaffolds Targeting the HIV Glycan Shield for an HIV Vaccine.

针对 HIV 疫苗的 HIV 聚糖屏蔽的基因支架。

• 批准号: 8877398
• 负责人: Yu Geng
• 金额: $ 30万
• 依托单位: PROSCI, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8877398
• 关键词: Acquired Immunodeficiency Syndrome; Address; Adjuvant; Animals; Antibodies; Antibody Affinity; Antibody Response; Antibody Specificity; Antigens; Binding; Bioinformatics; CD4 Positive T Lymphocytes; Carbohydrates; Cells; Complex; Dendritic Cells; Enzymes; Epitopes; Funding; Fungal Proteins; Genetic; Glycoproteins; Goals; HIV; HIV Envelope Protein gp120; HIV vaccine; HIV-1; Health; Human; Immune response; Immunization; Immunoglobulin Class Switching; Immunoglobulin G; Infection; Insecta; Lead; Link; Macaca; Mannose; Methods; Oryctolagus cuniculus; Patients; Phase; Plants; Polysaccharides; Process; Production; Proteins; Public Health; Saccharomyces cerevisiae; Series; Serum; Site; Solutions; Structure; Sum; System; Testing; Vaccines; Vertebral column; Viral; Virion; Virus; Yeasts; carbohydrate structure; density; design; env Gene Products; genome-wide analysis; glycosylation; immunogenicity; mannosyl(9)-N-acetylglucosamine2; mutant; neutralizing antibody; neutralizing monoclonal antibodies; novel; pandemic disease; prevent; product development; prophylactic; scaffold; simian human immunodeficiency virus; transmission process; yeast protein

DESCRIPTION: HIV-1/AIDS remains a serious public health problem globally with an estimated 7,000 new infections each day. An effective HIV-1 vaccine will be necessary for the long-term control of the HIV-1 pandemic. The goal of this study is to develop a safe and effective vaccine to prevent HIV-1 acquisition and transmission. Several potent and broadly neutralizing monoclonal antibodies (bNAbs) derived from HIV-1 infected patients bind to high mannose carbohydrate structures on the gp120 subunit of the HIV-1 envelope (Env) protein, such as 2G12 and a series of newly identified antibodies PGT125-131 and PGT135 that are among the most potent and broadly cross-reactive bNAbs identified to date. The identification of these bNAbs suggests that the glycan shield of gp120 is a viable vaccine target. We have targeted the HIV-1 Env glycan shield by genetically modifying a yeast strain and have shown that the elimination of three carbohydrate processing enzymes in S. cerevisiae (TM for triple mutant) results in the uniform production of the Man8GlcNAc2 carbohydrate structures that are the major form of glycans in the epitopes recognized by the PGT bNAbs and 2G12. We have shown that immunization of rabbits with whole TM yeast cells as well as purified, highly glycosylated, 2G12/PGT128 reactive TM yeast proteins, without the use of any gp120 or HIV-1 related proteins, elicits antibodies that recognize synthetic high-mannose glycans as well as monomeric gp120 proteins from a wide array of HIV-1 isolates and all tested virions in a carbohydrate dependent manner. The genetic scaffold elicited antibodies also potently neutralize HIV-1 from different subtypes that carry high-mannose N-glycans although potent neutralizing activity to primary virus strains has not been observed so far. We hypothesize that appropriate presentation of high mannose clusters on heterologous proteins in a combination with optimized immunization strategies can elicit antibodies that specifically target the key neutralizing epitope on the glycan shield of HIV-1 Env and potently neutralize primary isolates, and so propose the following Specific Aims: 1. Select lead immunogens that can efficiently support PGT bNAb binding from genome-wide screens of highly glycosylated proteins using a bioinformatics approach in combination with the genetically modified TM yeast for immunization trials. 2. Optimize immunization strategies and elicit neutralizing antibodies using a stepwise scaffold prime-Env boost strategy with the lead immunogens and a soluble HIV-1 gp140 trimer.

描述：HIV-1/AIDS在全球范围内仍然是一个严重的公共卫生问题，每天估计有7,000个新感染。长期控制HIV-1大流行将需要有效的HIV-1疫苗。这项研究的目的是开发一种安全有效的疫苗，以防止HIV-1采集和传播。源自HIV-1感染患者的几种有效且广泛中和的单克隆抗体（BNAB）与HIV-1 INVELOPE（INV）蛋白的GP120亚基上的高甘露糖碳水化合物结构结合，例如2G12，例如2G12，例如2G12，以及一系列新近识别的抗体PGT125-125-131和PGT135，迄今为止确定的BNAB。这些BNAB的鉴定表明GP120的聚糖屏蔽是可行的疫苗靶标。我们通过基因修饰酵母菌菌株来瞄准HIV-1 Envlycan屏蔽层，并表明在酿酒酵母中消除了三种碳水化合物加工酶（用于三突变体的TM）导致MAN8GLCNAC2碳水化合物结构均匀产生，这些结构是由glycans in 2 and epepts and pgtive and p.ab and p.ab and p.ab and ipopeopepopeospeopepopeps and ipopeps and ipopeps and ipopeps and ipopsips。我们已经表明，用整个TM酵母细胞的兔子免疫以及纯化的，高度糖基化的2G12/PGT128反应性TM酵母蛋白，而无需使用任何GP120或HIV-1相关蛋白，引发了从合成的高型gp120 and a inimane GP120抗体的抗体，并引发抗体。以碳水化合物依赖的方式测试了病毒体。遗传支架引起的抗体还可以从不同亚型中和携带高频含N-聚糖的HIV-1有效中和HIV-1，尽管到目前为止尚未观察到有效的对原发性病毒菌株的有效中和活性。我们假设在与优化的免疫策略结合使用的异源蛋白上适当呈现高甘露糖簇可以引起抗体，这些抗体特异性地针对HIV-1 Env的聚糖屏蔽层的钥匙中和表位，并在HIV-1 Env的Glycan Shield上进行了有效中和原发性分离株，因此提出了以下特定目标： 1。选择可以使用生物信息学方法与遗传改性的TM酵母进行免疫试验的生物信息学方法，从而有效地从高度糖基化蛋白的全基因组筛选中有效地支持PGT BNAB结合。 2.使用铅脚手架质量启动策略和可溶性HIV-1 GP140 Trimer，优化免疫策略并通过逐步脚手架质量增强策略引起中和抗体。