CD4 and C3d fusion proteins-antibodies to HIV-1 envelope

CD4 和 C3d 融合蛋白 - HIV-1 包膜抗体

• 批准号: 6532851
• 负责人: Ted M Ross
• 金额: $ 20.93万
• 依托单位: EAST CAROLINA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6532851
• 关键词: AIDS vaccines; CD4 molecule; HIV envelope protein; active immunization; antibody specificity; antibody titering; chimeric proteins; drug screening /evaluation; enzyme linked immunosorbent assay; humoral immunity; immunoglobulin G; laboratory rabbit; molecular cloning; neutralizing antibody; plasmids; vaccine development; vector vaccine; western blottings

DESCRIPTION (Provided by applicant): A central problem for the development of HIV-1 vaccines has been to identify immunogens capable of raising high titer, long lasting, neutralizing antibody to the envelope glycoproteins (Envs) of primary isolates. The HIV-1 Env has proven to be a weak immunogen, raising low titer antibodies that are slow to undergo affinity maturation. In this proposal, DNA immunogens will be used to test soluble forms of CD4 (sCD4) fused to Env in order to generate neutralizing antibodies to protected regions. In HIV/cell interactions, Env attaches to CD4 on the cell surface. This interaction leads to changes in Env, which expose cryptic regions important for Env binding to co-receptors and subsequent fusion and entry into human cells. DNA immunogens of sCD4/Env will allow for the stable exposure of Env regions for generation of neutralizing antibodies (Ab). In addition, in order to enhance the neutralizing antibody response, DNA immunogens will be tested containing sCD4/Env fused to C3d. HIV-1 Env has been a target for developing an effective vaccine. Our laboratory has successfully used the C3d component of complement as a molecular adjuvant for viral glycoproteins. In normal immune responses, the attachment of Gd to an antigen enhances both the initiation and the maturation of antigen-specific antibody. To test a potential role of sCD4 and C3d for Env, DNA constructs will be produced encoding Env fused to sCD4 (sCD4/Env) and sCD4/Env/C3d and used for immunizations. The study will be executed according to three specific aims. (1) Vaccine plasmids encoding a secreted monomeric (gpl20) form of primary Envs and these same forms of Env fused at the amino terminus with one of two forms for sCD4 (2 domain or 4 domain) and/or at the carboxyl terminus to three tandem copies of C3d will be constructed. The levels of expression and secretion of the various plasmid constructs will be determined. (2) The Env constructs will be compared for immunogenicity in rabbits, using DNA immunization. Raised antibody will be titered for the level of Env-specific IgG and for neutralizing activity for a panel of primary HIV-1 isolates. (3) Codon-optimized Env genes will be used in these same constructs and efficient expression and immunogenicity will be determined. Our goal is to identify the most favorable sCD4/Env/C3d fusion for raising high titer neutralizing antibody. The hypothesis throughout the study is that the sCD4 fusion will increase the neutralizing Ab response and the C3d fusion will enhance the immunogenicity of Env both by increasing the efficiency of the initiation of anti-Env Ab response and by increasing the ability of anti-Env Ab to undergo affinity maturation.

描述（由申请人提供）：开发的中心问题 HIV-1疫苗已经鉴定出能够提高高滴度的免疫原， 持久的，中和抗体的包膜糖蛋白（Envs） 主要分离物。HIV-1 Env已被证明是一种弱免疫原， 缓慢经历亲和力成熟的滴度抗体。在这 建议，DNA免疫原将用于测试可溶性形式的CD 4（sCD 4）融合 以产生针对受保护区域的中和抗体。在 在HIV/细胞相互作用中，Env附着在细胞表面的CD 4上。这 相互作用导致Env的变化，这暴露了重要的隐蔽区域， Env与辅助受体结合，随后融合并进入人细胞。 sCD 4/Env的DNA免疫原将允许Env区域的稳定暴露 用于产生中和抗体（Ab）。另外为了 增强中和抗体反应，DNA免疫原将被测试 含有与C3 d融合的sCD 4/Env。HIV-1 Env一直是开发 有效疫苗。我们的实验室已经成功地使用了C3 d组件， 补体作为病毒糖蛋白的分子佐剂。在正常免疫 在免疫应答中，Gd与抗原的结合增强了免疫应答的起始和 抗原特异性抗体的成熟。测试sCD 4的潜在作用 和C3 d用于Env，将产生编码与sCD 4融合的Env的DNA构建体 (sCD4/Env）和sCD 4/Env/C3 d，并用于免疫。本研究将 根据三个具体目标。(1)编码a的疫苗质粒 分泌的单体（gp 120）形式的初级Env和这些相同形式的Env 在氨基末端与sCD 4的两种形式之一融合（2结构域或4结构域 结构域）和/或在羧基末端与C3 d的三个串联拷贝连接， 构建了各种质粒的表达和分泌水平 将确定结构。(2)将比较Env结构， 免疫原性，使用DNA免疫。抗体升高将是 对Env特异性IgG的水平和中和活性进行滴定， 一组主要HIV-1分离株。(3)密码子优化的Env基因将用于 这些相同的构建体和有效的表达以及免疫原性将 测定我们的目标是确定最有利的sCD 4/Env/C3 d融合， 产生高滴度中和抗体。在整个研究中， sCD 4融合将增加中和抗体应答和C3 d 融合将增强Env的免疫原性， 启动抗Env Ab应答的能力，并通过增加 抗Env Ab经历亲和力成熟。

项目成果

Particle-based vaccines for HIV-1 infection.

用于 HIV-1 感染的颗粒疫苗。

• DOI: 10.2174/1568005033481213
• 发表时间: 2003
• 期刊: Current drug targets. Infectious disorders
• 作者: Young,KellyR;Ross,TedM
• 通讯作者: Ross,TedM

A minimum CR2 binding domain of C3d enhances immunity following vaccination.

C3d 的最小 CR2 结合域可增强疫苗接种后的免疫力。

• DOI: 10.1007/0-387-34134-x_17
• 发表时间: 2006
• 期刊: Advances in experimental medicine and biology
• 作者: Bower,JosephF;Ross,TedM
• 通讯作者: Ross,TedM

Characterization of a DNA vaccine expressing a human immunodeficiency virus-like particle.

表达人类免疫缺陷病毒样颗粒的 DNA 疫苗的表征。

• DOI: 10.1016/j.virol.2004.07.009
• 发表时间: 2004
• 期刊: Virology
• 影响因子: 3.7
• 作者: Young,KellyR;Smith,JamesM;Ross,TedM
• 通讯作者: Ross,TedM

Mouse strain-dependent differences in enhancement of immune responses by C3d.

C3d 增强免疫反应的小鼠品系依赖性差异。

• DOI: 10.1016/j.vaccine.2003.10.050
• 发表时间: 2004
• 期刊: Vaccine.
• 作者: Toapanta,FranklinR;Ross,TedM
• 通讯作者: Ross,TedM

Induction of heterosubtypic immunity to influenza A virus using a DNA vaccine expressing hemagglutinin-C3d fusion proteins.

使用表达血凝素-C3d 融合蛋白的 DNA 疫苗诱导针对甲型流感病毒的异亚型免疫。

• DOI: 10.1016/s0264-410x(02)00539-x
• 发表时间: 2003
• 期刊: Vaccine
• 影响因子: 5.5
• 作者: Mitchell,JudyA;Green,ThomasD;Bright,RickA;Ross,TedM
• 通讯作者: Ross,TedM