SINGLE CELL ANALYSIS OF A MULTI-TIER AIDS VACCINE

多层艾滋病疫苗的单细胞分析

• 批准号: 6167468
• 负责人: PETER C DOHERTY
• 金额: $ 20.79万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-15 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6167468
• 关键词: AIDS; AIDS vaccines; B lymphocyte; HIV envelope protein; T lymphocyte; cellular immunity; enzyme linked immunosorbent assay; human immunodeficiency virus 1; immunomodulators; laboratory mouse; leukocyte activation /transformation; recombinant virus; single cell analysis; tissue /cell culture; vaccine development; vaccinia virus; vector vaccine; virus DNA

The development of an effective HIV vaccine is perhaps the greatest challenge currently facing the biomedical research community. A consensus is emerging that an effective vaccine must activate both the humoral and cellular arms of the immune response and, in addition, take into account the tremendous variability among HIV isolates. The development and refinement of such a vaccine is crucially dependent on model systems in which specific B and T cell responses can be precisely quantified. Our preliminary studies show that a single HIV envelope elicits strong antibody responses in mice when administered first as DNA, then as recombinant vaccinia virus, and finally as purified protein (i.e., a D-V-P vaccination regimen). The overall aim of this project is to use a mouse model to comprehensively evaluate the D-V-P as a vehicle for the delivery of multiple HIV envelopes in a highly immunogenic form. We will apply methodologies that are well-established in our laboratory to measure envelope-specific B cell and CD4+ and CD9+ T cell responses at the single cell level. Initially, we will characterize specific cellular responses during the immunization of mice with single HIV envelopes using the D-V-P regimen. Two distinct, well characterized envelopes from the HIV-1 strains IIIB and MN will be used. The second part of this project will test the effectiveness of the D-V-P regimen in presenting multiple envelopes simultaneously to elicit a greater diversity of B and T cell responses. In the first experiments in this series, both IIIB and MN will be included in each step of the D-V-P regimen. We will then address the immunological consequences of administering IIIB or MN at only one or two of the vaccination steps. Finally, we will evaluate the response of mice to a proposed multi-envelope clinical vaccine based on the D-V-P regimen. These studies will provide information that is essential for the rational evaluation and refinement of multi-step, multi-vector vaccination strategies.

有效的HIV疫苗的开发可能是目前面临生物医学研究界面临的最大挑战。一定的共识是，有效的疫苗必须激活免疫反应的体液和细胞臂，此外，还考虑了HIV分离株之间的巨大变异性。这种疫苗的开发和改进至关重要地取决于模型系统，在该模型系统中，可以精确量化特定的B和T细胞反应。我们的初步研究表明，首先以DNA给药时，单个HIV包膜会在小鼠中引起强抗体反应，然后作为重组疫苗病毒，最后是纯化的蛋白质（即D-V-P-P疫苗接种方案）。该项目的总体目的是使用小鼠模型来全面评估D-V-P作为以高度免疫原性传递多种HIV信封的载体。我们将应用实验室中建立良好的方法来测量单个细胞水平的包膜特异性B细胞以及CD4+和CD9+ T细胞反应。最初，我们将在使用D-V-P方案的单个HIV信封对小鼠免疫过程中表征特定的细胞反应。将使用两个不同的，具有HIV-1菌株IIIB和MN的良好特征的信封。该项目的第二部分将测试D-V-P方案同时提出多个信封的有效性，以引起B和T细胞反应的更大多样性。在本系列的第一个实验中，IIIB和MN都将包括在D-V-P治疗方案的每个步骤中。然后，我们将仅在一个或两个疫苗接种步骤下管理IIIB或MN的免疫学后果。最后，我们将根据D-V-P方案评估小鼠对拟议的多层临床疫苗的反应。这些研究将提供对多步，多载体疫苗接种策略的合理评估和完善至关重要的信息。