HIV-1 DNA VACCINES THAT TARGET AND ACTIVATE CD40

针对并激活 CD40 的 HIV-1 DNA 疫苗

• 批准号: 6336090
• 负责人: JEFFREY A LEDBETTER
• 金额: $ 25.02万
• 依托单位: PACIFIC NORTHWEST RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6336090
• 关键词: AIDS vaccines; CD40 molecule; HIV envelope protein gp120; HIV envelope protein gp160; HIV envelope protein gp41; MHC class II antigen; Macaca nemestrina; antibody formation; antibody neutralization test; antibody titering; antigen presenting cell; cellular immunity; cytokine receptors; cytotoxic T lymphocyte; enzyme linked immunosorbent assay; helper T lymphocyte; human immunodeficiency virus 1; human tissue; humoral immunity; neutralizing antibody; plasmids; receptor expression; vector vaccine; virus antigen; virus protein

HIV-1 DNA vaccines induce both humoral and cellular immune responses in experimental animals, suggesting their potential usefulness for immunization. However, further optimization of DNA vaccines is needed to induce protection against virulent HIV-1 isolates. We propose to utilize CD40 activation and targeting in an attempt to improve the ability of HIV-1 env DNA vaccines to generate strong cellular immunity and high titers of neutralizing antibodies. We will construct DNA vaccines encoding CD40 ligand (CD154) or a single chain Fv (scFv) specific for CD40, fused with DNA encoding portions of the HIV-1 env protein. After DNA vaccination, the expression of this fusion protein in vivo could result in both activation of the CD40 receptor and direction of HIV-1 env antigens into the endocytic pathway of CD40 positive antigen presenting cells (APC). Internalization of env antigens after binding the CD40 positive antigens presenting cells (APC). Internalization of env antigens after binding the CD40 receptor will enhance presentation of peptides to effective antibody production and generation of CD4+ helper T cell and CD8+CTL activity. We hypothesize that the combination of CD40 activation and presentation of viral peptides by MHC class II will lead to improved immune responses to viral antigens. This hypothesis will be tested by construction of anti-human CD40 scFv-env DNA vaccines, followed by testing in primates where the anti-human CD40 scFv and human CD154 retain CD40 binding and functional activity. Antibody responses towards HIV-1 env will be tested by epitope specific ELISA assays and viral neutralization assays.

HIV-1 DNA 疫苗在实验动物中诱导体液和细胞免疫反应，表明它们在免疫方面具有潜在的用途。然而，需要进一步优化 DNA 疫苗以诱导针对有毒 HIV-1 分离株的保护。我们建议利用CD40激活和靶向来尝试提高HIV-1 env DNA疫苗产生强细胞免疫和高滴度中和抗体的能力。我们将构建编码 CD40 配体 (CD154) 或 CD40 特异性单链 Fv (scFv) 的 DNA 疫苗，并与编码 HIV-1 包膜蛋白部分的 DNA 融合。 DNA疫苗接种后，这种融合蛋白在体内的表达可以导致CD40受体的激活和HIV-1包膜抗原进入CD40阳性抗原呈递细胞（APC）的内吞途径的引导。结合 CD40 阳性抗原呈递细胞 (APC) 后，env 抗原内化。结合 CD40 受体后，env 抗原的内化将增强肽的呈递，从而有效产生抗体、产生 CD4+ 辅助 T 细胞和 CD8+CTL 活性。我们假设 CD40 激活和 MHC II 类病毒肽呈递的结合将导致对病毒抗原的免疫反应改善。这一假设将通过构建抗人 CD40 scFv-env DNA 疫苗进行测试，然后在灵长类动物中进行测试，其中抗人 CD40 scFv 和人 CD154 保留 CD40 结合和功能活性。针对 HIV-1 env 的抗体反应将通过表位特异性 ELISA 测定和病毒中和测定进行测试。