SHIV CHALLENGE & IN VIVO PASSAGE

艾滋病毒挑战

• 批准号: 6116329
• 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/6116329
• 关键词: AIDS; Mammalia; Primates; immunology; infection; inflammation; lymphatic system; model design /development; vaccines; virus

Two SHIV challenges have been studied in this project. The first involved 12 previously immunized animals. Six had been vaccinated with vaccinia virus expressing HIV-1LAI gp160 and subsequently boosted with purified subunit gp160 produced by vaccinia virus-infected AGMK cells. The other six served as mock-vaccinated controls, having been given vaccinia virus alone and boosted with IFA. Pre-virus challenge evaluation of the animals revealed serological responses similar to those achieved in humans immunized with various env preparations. The macaques were challenged with 25 tissue culture infectious doses (TCID50) of SHIVIIIB. The six mock-vaccinated animals became persistently viremic by virus culture and RNA PCR. In contrast, virus was recovered intermittently from one of the six vaccinated animals and the RNA PCR revealed virus levels several logs lower in the vaccinated animals. The animals will continue to be monitored and will be rechallenged with a more virulent SHIV. In the second challenge experiment, 22 M. nemestrina divided among 7 experimental vaccine and control groups also were challenged with SHIVIIIB. The results supported those in the first challenge experiment, that vaccinia env prime and env boost afforded the greatest protection against challenge. Two macaques, one from each control group, experienced spontaneous CD4 decline. To determine whether virus from these macaques had become more pathogenic, we gave whole-blood transfusions to two naive animals. The recipient macaques experienced a rapid and irreversible CD4 decline. Virus was recovered from the transfusion recipients and stocks were prepared. To confirm that the stocks were infectious in vivo, we inoculated two additional macaques with 1 ml of undiluted virus. These animals also showed CD4 depletion. The stock will be titered in additional macaques to determine the macaque infectious dose.

本项目研究了两个SHIV挑战。 第一次涉及12只先前免疫的动物。 其中六人接种了表达HIV-1 LAI gp 160的牛痘病毒，随后用牛痘病毒感染的AGMK细胞产生的纯化亚基gp 160进行加强免疫。 另外6只作为模拟接种对照组，仅给予牛痘病毒并用IFA加强。 动物的病毒攻击前评价显示，血清学应答与用各种env制剂免疫的人中获得的应答相似。 用25个组织培养感染剂量（TCID 50）的SHIVIIIB攻击猕猴。 通过病毒培养和RNA PCR，6只模拟接种动物出现持续病毒血症。 相比之下，从6只接种疫苗的动物中的一只间歇性回收病毒，RNA PCR显示接种疫苗的动物中的病毒水平低几个对数。 将继续监测动物，并使用毒性更强的SHIV进行再攻毒。 在第二次挑战实验中，22 M.将Nemestrina分成7个实验疫苗组和对照组，也用SHIVIIIB攻击。 结果支持了第一次攻击实验中的结果，即牛痘env初免和env加强提供了最大的对抗攻击的保护。 2只猕猴（每个对照组各1只）出现自发性CD 4下降。 为了确定来自这些猕猴的病毒是否变得更具致病性，我们给两只未感染过的动物输全血。 受体猕猴经历了快速和不可逆的CD 4下降。 从输血受者中回收病毒并制备贮备液。 为了确认储备液在体内具有感染性，我们用1 ml未稀释的病毒接种另外两只猕猴。 这些动物也显示出CD 4耗竭。 将在其他猕猴中对储备液进行滴度，以确定猕猴感染剂量。