FETAL IMMUNOPROPHYLAXIS AGAINST A LENTIVIRUS

胎儿针对慢病毒的免疫预防

• 批准号: 2069360
• 负责人: Ruth Margrit Ruprecht
• 金额: $ 37.73万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-04-01 至 1997-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2069360
• 关键词: Macaca mulatta; chimeric proteins; congenital infection; fusion gene; human immunodeficiency virus 1; immunopharmacology; immunotherapy; in situ hybridization; monoclonal antibody; neutralizing antibody; passive immunization; pharmacokinetics; placental transfer; polymerase chain reaction; simian immunodeficiency virus; tissue /cell culture

Using ultrasound (US)-guided inoculation of the amniotic fluid with the simian immunodeficiency virus (SIV), we have achieved a 86% fetal infection rate in rhesus monkeys (Macaca mulatta). However, neutralizing antibodies developed against HIV-1 cannot be evaluated in this system. Thus, we propose to establish a fetal infection model in macaques using a chimeric virus based on the infectious molecular clone SlVmac239 in which the tat, rev and env sequences of SIV were replaced by homologous sequences of HIV, and into which the vpu sequences of HIV were inserted as welL The resulting chimeric virus, SHIV-vpu+, is infectious in rhesus monkeys. The Specific Aims for this proposal are to: 1. Titrate the dose of SHIV-vpu+ required for reproducible fetal infection via the amniotic fluid. using 8 pregnant M. mulatta late in gestation. The mother/infant pairs will be followed for viremia and disease. 2. Examine a panel of neutralizing human anti-HIV-1 Env monoclonal antibodies (mAbs) for their ability to neutralize SHIV-vpu+. Human mAbs directed against the CD4 binding domain or against the V3 loop will be examined for their activity against SHIV-vpu+ in vitro, singly and in combination regimens. 3. Determine the pharmacokinetics of human mAbs during gestation by weekly injections into pregnant, uninfected macaques. Neutralizing titers in serum, amniotic fluid, cord blood, placental tissues and milk will be determined. Mothers and infants will be examined carefully for signs of toxicity. 4. Test passive immunoprophylaxis with the optimal combination regimen of human anti-HIV mAbs (as determined in Specific Aim #2) during gestation. We anticipate that a combination of mAbs directed against the CD4 binding domain and the V3 loop will be used. Pregnant controls will be treated with irrelevant human mAbs. After pre-treatment, all animals will be challenged with SHIV-vpu + via the amniotic fluid. Moab therapy will be continued until Cesarean section. Mother/infant pairs will be followed for development of viremia and disease. The proposed work is highly significant because it allows in vivo analysis of neutralizing human Moab's directed against the HIV-1 envelope glycoprotein.

使用超声（US）引导的羊水接种 猿猴免疫缺陷病毒（SIV），我们达到了86％的胎儿 恒河猴（Macaca mulatta）的感染率。 但是，中和 在该系统中无法评估针对HIV-1开发的抗体。 因此，我们建议使用猕猴建立胎儿感染模型 基于感染性分子克隆SLVMAC239的嵌合病毒 SIV的TAT，REV和ENV序列被同源 HIV的序列以及插入HIV的VPU序列 以及所得的嵌合病毒Shiv-VPU+在恒河猴中具有感染性 猴子。 该提案的具体目的是： 1。滴定可再现胎儿所需的Shiv-VPU+剂量 通过羊水感染。 在妊娠后期使用8个怀孕的M. Mulatta。母亲/婴儿对 病毒血症和疾病将受到遵循。 2。检查一个中和人类抗HIV-1 env单克隆的面板 抗体（mAb）的中和湿vpu+的能力。人物mab 针对CD4结合域或针对V3环的针对V3环路将是 检查了它们对Shiv-VPU+体外，单独和IN的活动 组合方案。 3。通过 每周注射未感染的怀孕，未感染的猕猴。 中和 血清中的滴度，羊水，脐带血，胎盘组织和牛奶 将确定。 将仔细检查母亲和婴儿 毒性的迹象。 4。使用最佳组合方案测试被动免疫预防 人类抗HIV mab（如特定目的＃2中确定） 妊娠。我们预计将对MAB的组合针对 CD4结合域和V3环将使用。怀孕的对照将 用无关的人物mab处理。预处理后，所有动物 SHIV-VPU +将通过羊水挑战。摩押治疗 将继续持续到剖宫产。母/婴儿对 紧随其后的病毒血症和疾病的发展。 拟议的工作非常重要，因为它允许体内 分析中和针对HIV-1信封的人类摩押 糖蛋白。