VACCINE REGIMENS TO INDUCE CD4+ AND CD8+ T CELLS AGAINST SIV EPITOPES

诱导 CD4 和 CD8 T 细胞对抗 SIV 表位的疫苗方案

• 批准号: 8173128
• 负责人: David I Watkins
• 金额: $ 5.16万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173128
• 关键词: Adenovirus Vector; Adenoviruses; Animals; Autologous; Autologous Dendritic Cells; Basic Science; CD4 Positive T Lymphocytes; CD8B1 gene; Cell Separation; Cells; Clinical; Computer Retrieval of Information on Scientific Projects Database; DNA; Dendritic Cells; Dose; Epitopes; Failure; Funding; Genes; Grant; HIV; HIV vaccine; Health Services Research; Hepatitis; Hepatitis B Core Antigen; Immunization; Individual; Institution; Laboratories; Methods; Peptides; Peripheral Blood Mononuclear Cell; Phase; Physiologic pulse; Protocols documentation; Regimen; Research; Research Personnel; Resources; Role; SIV; Services; Source; T cell response; T-Lymphocyte; Testing; United States National Institutes of Health; Vaccinated; Vaccination; Vaccines; Virus; cost; enzyme linked immunospot assay; gene discovery; gene gun; novel; novel vaccines; vector; vector vaccine; vector-based vaccine

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Objective: To compare four novel vaccine regimens to induce CD4+ and CD8+ T Cells against SIV epitopes. Recent failures in clinical HIV vaccines have underscored the importance of more thoroughly evaluating basic science of HIV as well as testing new vaccine regimens and vectors. In an effort to overcome the limitations of more traditional vector-based vaccines, our laboratory has developed several novel immunization strategies. These new methods will allow us to directly prime specific T cell responses in a manner that previously has been impossible with other vaccine vectors and regimens. This should allow us to dissect the contributions of specific T cell responses in the control of SIV replication- the roles of both subdominant CD8+ T cells and virus-specific CD4+ T cells. In the R21 phase of this grant, we will compare four novel vaccination regimens: peptide-pulsed dendritic cells, peptide-conjugated nanobeads, peptide-pulsed PBMC, and SIV peptides fused to a Hepatitis B core antigen (HBcAg) carrier gene. SUBPROJECT PROGRESS: We have completed vaccinating animals in groups one and two of the R21 phase. These animals received autologous dendritic cells pulsed with three Mamu-DR¿w*606-restricted CD4 epitopes and primed with either autologous PBMC pulsed with the same epitopes or peptide-conjugated nanobeads. Two weeks after the final boosts, no animals made any detectable epitope-specific CD4+ T cell responses as detected by ELISPOT or ICS. The HBcAg vectors were completed by Dr. Deborah Fuller with each of the selected Mamu-A*01-restricted CD8 epitopes inserted into individual vectors. Six Mamu-A*01+ animals, groups three and four, received four doses of the HBcAg vector; however, unfortunately it was later discovered that the gene gun administering these vaccines was not optimally functioning. We're going to give all six animals a fifth HBcAg dose. After the fourth dose, no animals made any detectable epitope-specific CD8+ T cell response; however, they did respond well to Hepatitis peptide provided by Dr. Fuller indicating that they did receive some doses of the vaccine. Once given a fifth dose, we will move into the boost of these groups. One group will receive autologous PBMC pulsed with each of the A*01 epitopes, while the other group will receive the A*01 epitopes conjugated to nanobeads. We will request a one year no-cost extension for this project. For the CD4 Aim of the R21, we will repeat the dendritic cell prime-nanobead boost using a longer dendritic cell isolation protocol. Additionally, we will make DNA and Adenovirus vectors containing the three CD4 epitopes. We will vaccinate animals with these constructs in the hope of eliciting epitope-specific CD4+ T cell responses. Lastly, if the HBcAg prime-PBMC/nanobead boost regimens are unsuccessful, we will also DNA prime-Adenovirus boost animals with the A*01 epitopes to successfully elicit epitope-specific CD8+ T cell responses. This research uses Animal Services and Research Services.

该副本是使用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这是调查员的机构。 目的：比较四种新型疫苗方案以诱导CD4+和CD8+ T细胞针对SIV表位。 临床HIV疫苗的最新失败促进了更彻底评估HIV的基础科学以及测试新的疫苗方案和载体的重要性。为了克服更传统的基于矢量的疫苗的局限性，我们的实验室制定了几种新型的免疫抑制策略。这些新方法将使我们能够以其他疫苗向量和方案不可能以先前是不可能的方式直接启动特定的T细胞反应。这应该使我们能够剖析特定T细胞反应在控制SIV复制中的贡献 - 亚抑制CD8+ T细胞和病毒特异性CD4+ T细胞的作用。 在该赠款的R21阶段中，我们将比较四种新型的疫苗接种方案：辣椒脉冲的树突状细胞，辣椒偶联的纳米棒，辣椒粉的PBMC和SIV Pepperides融合到肝炎B核B核B核B核抗原（HBCAG）携带者基因中。 subproject进度： 我们已经在R21期的第一组和第二组中完成了接种疫苗的动物。这些动物接受了用三个Mamu-dr脉冲的自体树突状细胞，其限制性CD4表位，并用具有相同表位的自体PBMC或肽偶联的纳米珠脉冲。最终提升后两周，没有动物使ELISPOT或ICS检测到的任何可检测的表位特异性CD4+ T细胞反应。 HBCAG矢量由Deborah Fuller博士完成，每个选定的Mamu-A*01限制的CD8表位插入了单个向量。六个Mamu-A*01+动物，第三组和四组，接受了四剂HBCAG载体；但是，不幸的是，后来发现给予这些疫苗的基因枪并非最佳功能。我们将向所有六只动物提供第五个HBCAG剂量。第四剂量后，没有动物使任何可检测的表位特异性CD8+ T细胞反应。但是，他们确实对富勒博士提供的肽肽的反应很好，表明他们确实接受了一些剂量的疫苗。一旦服用了第五剂，我们将进入这些群体的提升。一组将接收带有A*01表位的自体PBMC，而另一组将接收到与纳米角共轭的A*01表位。 我们将要求为该项目提供一年的无成本延期。对于R21的CD4目的，我们将使用更长的树突状细胞分离方案重复树突状细胞Prime-Nanobead增强。此外，我们将制造包含三个CD4表位的DNA和腺病毒载体。我们将用这些构建体接种动物，以期引起表位特异性CD4+ T细胞反应。最后，如果HBCAG Prime-PBMC/Nanobead增强方案不成功，我们还将DNA Prime-AdeNovirus带有A*01表位的DNA Prime-AdeNovirus促进动物，以成功引起表位特异性CD8+ T细胞反应。 该研究使用动物服务和研究服务。