RESPIRATORY SYNCYTIAL VIRUS EFFICACY STUDY IN AFRICAN GREEN MONKEYS

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 在非洲绿猴 (AGM) 中进行了一系列三项研究，以测试 Tibotec Pharmaceuticals 配制的专利化合物 TMC 353121 对非人类灵长类动物中 RSV 的抵抗力。在之前的研究中，该化合物（一种 RSV 融合抑制剂）被证明对小型动物体内的 RSV 具有功效。这些研究是与 Bioqual, Inc. 合作进行的，动物和兽医人员在其工厂进行。我的实验室不仅要提供病毒挑战储备液，还要对三项研究中每项研究期间从动物身上采集的样本进行所有病毒学测试。我们对 48 只 AGM 进行了血清学检测，以预筛选并鉴定出 36 只 RSV 血清阴性动物用于研究。第一项研究 NC282 是一项感染研究，采用了 6 次 AGM。 我们制备了 37 瓶 1ml 的 RSV 菌株，两种浓度分别为 1x10e3pfu/ml 和 1x10e4pfu/ml，并在整个研究中冷冻保存。将动物分为两组，每组通过鼻内和气管内给药接受10e3pfu/ml (Gp.1)，另一组通过类似给药接受10e4pfu/ml (Gp.2)。每天采集鼻子和喉咙样本，每两天采集支气管肺泡灌洗样本，监测动物RSV感染的临床症状，并通过噬菌斑测定检测病毒载量。两组的病毒载量均在第 6 天达到峰值，在咽喉中为 3.7e2 pfu/ml 和 1.4 e3pfu/ml，在 BAL 中为 2e2 pfu/ml 和 3.7e2 pfu/ml，Gps1 和 2 分别达到峰值病毒载量。这些结果证实了该模型，并决定在随后的两项功效研究中使用 10e4pfu/ml 的接种剂量。 进行了两项连续静脉输注该化合物的功效研究。 研究 2 将 15 只动物分成三组，并配备 Gps。图1和2，以50ng/ml血浆水平施用TMC 353121的治疗和预防组，以及媒介物对照组Gp 3；研究 3 将 12 只动物分为三组，其中 Gps 1 和 2 治疗组仅以 5ng/ml 或 500ng/ml 的血浆水平施用 TMC 353121，Gp 3 为载体对照组。 对动物进行预处理，穿上夹克和系绳系统，接受植入导管的手术，并开始输注。动物研究最近已经完成，但是，我们仍在进行斑块和抗体测定并汇编数据。

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. A series of three studies were conducted in African green monkeys (AGM) to test a proprietary compound, TMC 353121, formulated by Tibotec Pharmaceuticals against RSV in nonhuman primates. In previous studies, the compound, an RSV fusion inhibitor was shown to have efficacy against RSV in vivo in smaller animals. These studies were performed in collaboration with Bioqual, Inc. with animals and veterinary personnel at their facility. My laboratory was to provide not only virus challenge stock but performed all virological testing of samples taken from the animals during each of the three studies. We performed serological testing of 48 AGMs to prescreen and identify 36 RSV seronegative animals to be used for the studies. The first study, NC282, was an infection study and utilized 6 AGM. We prepared 37, 1ml vials of RSV strain at each of two concentrations of 1x10e3pfu/ml and 1x10e4pfu/ml and cryopreserved them for the entire study. The animals were divided into two groups with each group receiving either 10e3pfu/ml (Gp.1) by intranasal and intratrachael administration and the other receiving 10e4pfu/ml (Gp.2) by similar administration. Animals were monitored for clinical signs of RSV infection with nose and throat samples taken daily and bronchoalveolar lavage samples taken every two days and tested for virus loads by plaque assay. Peak viral loads were reached in both groups by day 6 in the throat with 3.7e2 pfu/ml and 1.4 e3pfu/ml and in the BAL with 2e2 pfu/ml and 3.7e2 pfu/ml and for Gps1 and 2 respectively. These results confirmed the model and the decision to utilize the 10e4pfu/ml inoculation dose for the two subsequent efficacy studies was made. Two efficacy studies with the continuous intravenous infusion of the compound were performed. Study 2 had 15 animals divided into three groups with Gps. 1 and 2, therapeutic and prophylactic arms with TMC 353121 was administered at a plasma level of 50ng/ml and Gp 3 the vehicle control arms; and Study 3 had 12 animals divided into three groups with Gps 1 and 2 therapeutic arms only with TMC 353121 administered at plasma levels of either 5ng/ml or 500ng/ml and Gp 3 a vehical control group. Animals were preconditioned to wear the jacket and tethering system, underwent surgery for implanting the catheter, and infusions begun. The animal studies have recently been completed, however, we are still conducting plaque and antibody assays and compiling data.