Department of Microbiology & Immunology, University of Miami Miller School of Medicine, Miami, FL, USA.
Retrovirology. 2011 May 18;8:39. doi: 10.1186/1742-4690-8-39.
Molecular adjuvants are a promising method to enhance virus-specific immune responses and protect against HIV-1 infection. Immune activation by ligands for receptors such as CD40 can induce dendritic cell activation and maturation. Here we explore the incorporation of two CD40 mimics, Epstein Barr Virus gene LMP1 or an LMP1-CD40 chimera, into a strain of SIV that was engineered to be limited to a single cycle of infection.
Full length LMP1 or the chimeric protein LMP1-CD40 was cloned into the nef-locus of single-cycle SIV. Human and Macaque monocyte derived macrophages and DC were infected with these viruses. Infected cells were analyzed for activation surface markers by flow cytometry. Cells were also analyzed for secretion of pro-inflammatory cytokines IL-1β, IL-6, IL-8, IL-12p70 and TNF by cytometric bead array.
Overall, single-cycle SIV expressing LMP1 and LMP1-CD40 produced a broad and potent T(H)1-biased immune response in human as well as rhesus macaque macrophages and DC when compared with control virus. Single-cycle SIV-LMP1 also enhanced antigen presentation by lentiviral vector vaccines, suggesting that LMP1-mediated immune activation may enhance lentiviral vector vaccines against HIV-1.
分子佐剂是一种很有前途的方法，可以增强病毒特异性免疫反应，预防 HIV-1 感染。配体与受体（如 CD40）的相互作用可以诱导树突状细胞的激活和成熟。在这里，我们研究了将两种 CD40 模拟物，Epstein Barr 病毒基因 LMP1 或 LMP1-CD40 嵌合体，掺入到一种经过工程改造的只能进行单轮感染的 SIV 株中。
全长 LMP1 或嵌合蛋白 LMP1-CD40 被克隆到单循环 SIV 的 nef 基因座中。用人和猕猴单核细胞衍生的巨噬细胞和 DC 感染这些病毒。通过流式细胞术分析感染细胞的激活表面标志物。通过细胞因子微珠阵列分析细胞分泌促炎细胞因子 IL-1β、IL-6、IL-8、IL-12p70 和 TNF。
总的来说，与对照病毒相比，表达 LMP1 和 LMP1-CD40 的单循环 SIV 在人源和恒河猴巨噬细胞和 DC 中产生了广泛而有力的 T(H)1 偏向免疫反应。单循环 SIV-LMP1 还增强了慢病毒载体疫苗的抗原呈递，表明 LMP1 介导的免疫激活可能增强针对 HIV-1 的慢病毒载体疫苗。
