IMMUNIZATION W/ DNA CODING

DNA 编码免疫

• 批准号: 6116338
• 负责人: William Randall Morton
• 金额: $ 8.67万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6116338
• 关键词: AIDS; Mammalia; Primates; biotechnology; immunology; infection; inflammation; lymphatic system; model design /development; technology /technique; vaccines; virus

The gag proteins of SIV include the p8 nucleocapsid protein, which contains two cysteine arrays that are involved in viral RNA packaging and in a post-penetration step. Mutations in these arrays render the virus non-infectious. A deletion mutant (M14) of a molecular clone of SIVMne CL8 has been constructed that has a four amino acid deletion in the second cysteine array. This study is designed to test the efficacy of DNA immunizations in the SIV macaque model by immunizing macaques with DNA from the noninfectious viral construct, M14, as a potential AIDS vaccine. Five animals received mutant virus DNA and four controls were injected with vector plasmid DNA lacking the SIV construct. By SIV ELISA, antibody was detected after DNA immunizations, indicating that the mutant proviral DNA was expressed. None of the DNA-inoculated animals showed any signs of a productive SIV infection, indicating that the DNA construct is safe and does not lead to an established infection. Upon challenge, all four control animals became infected. Plasma virus was readily detected by QC-PCR assays at high levels (105 106 genome equivalents/ml) and virus was readily isolated from all four of these control animals. Of the five DNA-immunized animals, no virus was detected in one animal by either virus isolation or QC-PCR. Three of the animals became infected but the virus levels were less than in controls as determined by virus isolation and plasma RNA assays. Virus loads in one immunized animal were indistinguishable from those in control animals. One control animal and one immunized animal developed clinical AIDS and were euthanized in December 1997. The remaining animals are being monitored monthly.

SIV的GAG蛋白包括P8 Nucleocapsid蛋白，其中包含两个与病毒RNA包装有关的半胱氨酸阵列以及在渗透后步骤中。 这些阵列中的突变使病毒不感染。 已经构建了SIVMNE CL8的分子克隆的缺失突变体（M14），该突变体在第二个半胱氨酸阵列中具有四个氨基酸缺失。 这项研究旨在通过从非感染病毒构建体M14中免疫猕猴作为潜在的AIDS疫苗来测试SIV猕猴模型中DNA免疫的功效。 五只动物接受了突变病毒DNA，并注入了缺乏SIV构建体的载体质粒DNA。 通过SIV ELISA，在DNA免疫后检测到抗体，表明表达突变的病毒DNA。 DNA接种的动物均未显示出生产性SIV感染的任何迹象，表明DNA构建体是安全的，并且不会导致已建立的感染。 在挑战时，所有四只控制动物都被感染了。 通过QC-PCR分析很容易在高水平（105 106个基因组等效物/mL）中检测到血浆病毒，并且很容易从所有四种对照动物中分离出病毒。 在五只DNA免疫动物中，病毒分离或QC-PCR在一种动物中未检测到病毒。 这三只动物被感染，但病毒水平小于对照组，通过病毒分离和血浆RNA分析确定。 一种免疫动物中的病毒负荷与对照动物中的病毒负荷没有区别。 一只对照动物和一只免疫动物开发了临床艾滋病，并于1997年12月被安乐死。其余动物每月受到监测。