CD40 LIGAND AS ADJUVANT FOR RAISING PROTECTIVE ANTI-HIV AB BY DNA/MVA VACCINES

CD40 配体作为 DNA/MVA 疫苗产生保护性抗 HIV AB 的佐剂

• 批准号: 7715809
• 负责人: Liang-Chuan Lai
• 金额: $ 6.8万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7715809
• 关键词: Adjuvant; B-Lymphocytes; Binding; CD40 Ligand; CD80 gene; Cellular Immunity; Computer Retrieval of Information on Scientific Projects Database; DNA Vaccines; DNA/MVA vaccine; Extracellular Protein; Flow Cytometry; Funding; Grant; HIV; Institution; Interleukin-12; Macaca; Membrane; Modeling; Plasmids; Proteins; Research; Research Personnel; Resources; SIV; Source; TNFSF5 gene; Testing; United States National Institutes of Health; Virus-like particle; Western Blotting; research study

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. This project seeks to evaluate the adjuvant effects of CD40L to enhance the human immunodeficiency virus (HIV)-specific humoral and cellular immunity elicited by a DNA/MVA vaccine using a macaque/simian immunodeficiency virus (SIV) model. The macaque CD40L was cloned and expressed in cis (on the same plasmid) and in trans (on a separate plasmid) with respect to the SIV 239 vaccine DNA. In the cis form, the protein was expressed as a membrane bound form that is incorporated into SIV virus-like particles. In the trans form, the protein was expressed as a soluble secreted form. Intracellular expression of the CD40L protein was confirmed by flow cytometry and extracellular protein was confirmed by Western blotting. The expressed proteins were highly active in stimulating DC from SIV-infected macaques. These activated DC upregulated CD80 and produced TNF-a and IL-12. In addition, these proteins were active in stimulating macaque B cells. On going experiments are testing the adjuvant effects of these constructs in macaques.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 本项目旨在使用猕猴/猿猴免疫缺陷病毒（SIV）模型评价CD 40 L增强DNA/MVA疫苗诱导的人类免疫缺陷病毒（HIV）特异性体液和细胞免疫的佐剂效应。 克隆猕猴CD 40 L，并相对于SIV 239疫苗DNA以顺式（在同一质粒上）和反式（在单独的质粒上）表达。在顺式形式中，蛋白质表达为膜结合形式，其掺入SIV病毒样颗粒中。在反式形式中，蛋白质表达为可溶性分泌形式。通过流式细胞术证实CD 40 L蛋白的细胞内表达，通过Western印迹证实细胞外蛋白。 表达的蛋白在刺激来自SIV感染的猕猴的DC中具有高度活性。这些活化的DC上调CD 80并产生TNF-α和IL-12。此外，这些蛋白质在刺激猕猴B细胞中具有活性。正在进行的实验是测试这些结构在猕猴中的佐剂作用。