Potent Phage T4 Derived V2 Immunogens as HIV Vaccines

有效的噬菌体 T4 衍生 V2 免疫原作为 HIV 疫苗

• 批准号: 8868023
• 负责人: Venigalla B. Rao
• 金额: $ 47.13万
• 依托单位: CATHOLIC UNIVERSITY OF AMERICA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8868023
• 关键词: Acute; Adjuvant; Affinity; Antibodies; Antigens; Bacteriophage T4; Bacteriophages; Binding; Biological Assay; Biological Process; Blocking Antibodies; CCR5 gene; Capsid; Capsid Proteins; Cells; Chinese Hamster Ovary Cell; Clinical Trials; Dendritic Cells; Development; Engineering; Enterotoxins; Epitopes; Exposure to; Goals; HIV; HIV vaccine; HIV-1; Heating; Immune response; In Vitro; Incubated; Integrins; Intramuscular; Length; Libraries; Ligands; Membrane; Modeling; Molecular Conformation; Molecular Genetics; Monoclonal Antibodies; Mus; Mutation; Oryctolagus cuniculus; Pattern; Phase; Play; Protein Binding; Proteins; Reporting; Route; Sexual Transmission; Site; Skin; Specificity; Surface; System; T-Lymphocyte; Testing; Time; Transplantation; Vaccination; Vaccines; Variant; Viral Genome; Virus; base; fitness; glycosylation; high throughput screening; immunogenicity; nanoparticle; neutralizing antibody; nonhuman primate; novel; particle; prevent; prophylactic; scaffold; secretory protein; transmission process; vaccine candidate; vaccine delivery; vaccine development; vaccine trial; virus host interaction

DESCRIPTION (provided by applicant): A major goal of this proposal is to develop a novel experimental paradigm to engineer preventative HIV-1 vaccines. The V1/V2 domain of HIV-1 envelope is an important target for vaccine development for several reasons. V2 interaction with the �4�7 integrin of mucosal T cells may play a key role in the efficient capture of HIV-1 virus at the site of initial exposure during sexual transmission. The V1/V2 domain is also a key player in regulating neutralization sensitivity of HIV-1, and consists of epitopes recognized by some of the most potent broadly neutralizing antibodies reported to date. Here, we hypothesize that induction of antibodies by vaccination with V1/V2 immunogens that interferes, specifically with the V2 �4�7 interactions, confers protection against HIV-1 acquisition. Since the conformation of V2 is highly variable, the V2 domain will be constrained by transplanting it into the compactly folded 9 kDa small outer capsid protein (Soc) from bacteriophage T4. To select for the functionally important V2 conformations, libraries of V2 domain variants consisting of founder/acute virus sequences, different glycosylation patterns, and random mutations will be constructed and expressed in CHO cells as secretory proteins. High throughput assays will be developed, one to select �4�7 binding conformations and another to select V2 conformational antibody binding variants. The biologically active Soc-V2 conformational variants will be arrayed on phage T4 nanoparticles by incubating the purified Soc-V2 proteins with Soc-minus Hoc-minus phage, up to 870 copies per particle. A Hoc- fused targeting ligand such as Dec205 mAb will also be assembled on the same capsid, up to 155 copies per capsid, to target the T4-V2 nanoparticles to the antigen-presenting dendritic cells. The immunogenicity of T4-V2 nanoparticles will be evaluated in mice by intramuscular route as well as transcutaneous (skin) route using heat labile enterotoxin as an adjuvant. The immune responses will be quantified for V2- �4�7 interfering antibodies, virus neutralizing antibodies and transmission-blocking antibodies. The best V2 variants down-selected from these assays will then be tested in the rabbit model. A novel HIV-1 transmission assay will be developed, which will precisely quantify the number of virus genomes that cross the host membrane after a few minutes of exposure to CD4+, �4�7+ and CCR5+ T cells. Based on real-time PCR, this assay would be extremely sensitive, rapid, and high throughput. Using this assay, V2 variants that can induce antibodies which block virus entry by interfering with the initial virus-host interactions will be selected. These might potentially serve as preventative HIV-1 vaccine candidates for further studies in nonhuman primates and clinical trials.

描述（由申请人提供）：该提案的一个主要目标是开发一种新的实验范例来设计预防性 HIV-1 疫苗。出于多种原因，HIV-1 包膜的 V1/V2 结构域是 V2 疫苗开发的重要目标。与粘膜T细胞的�4�7整合素的相互作用可能在有效捕获HIV-1病毒的过程中发挥关键作用 V1/V2 结构域也是调节 HIV-1 中和敏感性的关键因素，由迄今为止报道的一些最有效的广泛中和抗体所识别的表位组成。通过接种 V1/V2 免疫原诱导产生抗体，特别是干扰 V2�4�7 相互作用，从而提供针对 HIV-1 感染的保护。由于 V2 的构象高度可变，因此 V2 结构域将受到以下因素的限制。将其移植到来自噬菌体 T4 的紧凑折叠的 9 kDa 小外衣壳蛋白 (Soc) 中 为了选择功能上重要的 V2 构象，将构建由创始人/急性病毒序列、不同糖基化模式和随机突变组成的 V2 结构域变体文库。将开发作为分泌蛋白在 CHO 细胞中构建和表达的高通量，一种用于选择分析 �4�7 结合构象，另一种用于选择 V2 构象抗体结合变体。通过将纯化的 Soc-V2 蛋白与 Soc-minus Hoc-minus 噬菌体一起孵育，活性 Soc-V2 构象变体将排列在噬菌体 T4 纳米颗粒上，每个颗粒最多 870 个拷贝。组装在同一衣壳上，每个衣壳最多 155 个拷贝，以将 T4​​-V2 纳米颗粒靶向抗原呈递将使用热不稳定肠毒素作为佐剂，通过肌内途径和经皮（皮肤）途径评估 T4-V2 纳米颗粒的免疫原性，并对 V2-4�7 干扰抗体的免疫反应进行量化。然后，将从这些测定中筛选出的最佳 V2 变体在兔子模型中进行测试。正在开发，它将精确量化暴露于 CD4+、�4�7+ 和 CCR5+ T 细胞几分钟后穿过宿主细胞膜的病毒基因组的数量。基于实时 PCR，该测定将极其灵敏。使用这种检测方法，将选择能够诱导抗体的 V2 变体，这些抗体通过干扰最初的病毒与宿主的相互作用来阻止病毒进入，这些变体可能作为预防性 HIV-1 疫苗候选物，用于非人类灵长类动物的进一步研究。和临床试验。

项目成果

A sequestered fusion peptide in the structure of an HIV-1 transmitted founder envelope trimer.

HIV-1 传播的创始人包膜三聚体结构中的隔离融合肽。

• 发表时间: 2019
• 影响因子: 16.6
• 作者: Ananthaswamy, Neeti;Fang, Qianglin;AlSalmi, Wadad;Jain, Swati;Chen, Zhenguo;Klose, Thomas;Sun, Yingyuan;Liu, Yue;Mahalingam, Marthandan;Chand, Subhash;Tovanabutra, Sodsai;Robb, Merlin L;Rossmann, Michael G;Rao, Venigalla B
• 通讯作者: Rao, Venigalla B

Glycosylation and oligomeric state of envelope protein might influence HIV-1 virion capture by α4β7 integrin.

包膜蛋白的糖基化和寡聚状态可能会影响α4β7 整合素的HIV-1 病毒颗粒捕获。

• 发表时间: 2017
• 影响因子: 3.7
• 作者: Chand, Subhash;Messina, Emily L;AlSalmi, Wadad;Ananthaswamy, Neeti;Gao, Guofen;Uritskiy, Gherman;Padilla;Mahalingam, Marthandan;Peachman, Kristina K;Robb, Merlin L;Rao, Mangala;Rao, Venigalla B
• 通讯作者: Rao, Venigalla B

The role of Siglec-1 in HIV-1/macrophage interaction.

Siglec-1 在 HIV-1/巨噬细胞相互作用中的作用。

• DOI: 10.14800/macrophage.1435
• 发表时间: 2016-10-17
• 期刊: Macrophage
• 作者: O. Jobe;Jiae Kim;M. Rao
• 通讯作者: M. Rao

TFH cells accumulate in mucosal tissues of humanized-DRAG mice and are highly permissive to HIV-1.

TFH 细胞在人源化 DRAG 小鼠的粘膜组织中积聚，并且对 HIV-1 高度敏感。

• 发表时间: 2015-06-02
• 影响因子: 4.6
• 作者: Allam, Atef;Majji, Sai;Peachman, Kristina;Jagodzinski, Linda;Kim, Jiae;Ratto;Wijayalath, Wathsala;Merbah, Melanie;Kim, Jerome H;Michael, Nelson L;Alving, Carl R;Casares, Sofia;Rao, Mangala
• 通讯作者: Rao, Mangala

Designing a soluble near full-length HIV-1 gp41 trimer.

设计可溶性近全长 HIV-1 gp41 三聚体。

• 发表时间: 2013-01-04
• 作者: Gao, Guofen;Wieczorek, Lindsay;Peachman, Kristina K;Polonis, Victoria R;Alving, Carl R;Rao, Mangala;Rao, Venigalla B
• 通讯作者: Rao, Venigalla B