RESPIRATORY SYNCYTIAL VIRUS EFFICACY STUDY IN AFRICAN GREEN MONKEYS

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 在非洲绿色猴（AGM）中进行了一系列三项研究，以检测Tibotec Pharmaceuticals配制的专利化合物TMC 353121在非人灵长类动物中抗RSV的作用。在先前的研究中，该化合物（RSV融合抑制剂）显示出在较小动物体内对RSV具有功效。这些研究与Bioqual，Inc.合作进行。与动物和兽医人员在他们的设施。我的实验室不仅提供病毒攻毒储备液，还对三项研究中的每项研究期间从动物中采集的样本进行所有病毒学检测。我们对48只AGM进行了血清学检测，以预筛选并确定36只RSV血清阴性动物用于研究。第一项研究NC 282是一项感染研究，使用了6个AGM。 我们制备了37个1 ml RSV毒株小瓶，浓度分别为1 × 10 e3 pfu/ml和1 × 10 e4 pfu/ml，并将其冷冻保存，用于整个研究。将动物分为两组，每组通过鼻内和鼻内给药接受10 e3 pfu/ml（Gp.1），另一组通过类似给药接受10 e4 pfu/ml（Gp.2）。通过每天采集鼻和咽喉样本以及每两天采集支气管肺泡灌洗样本监测动物的RSV感染临床体征，并通过空斑试验检测病毒载量。两组在第6天达到峰值病毒载量，咽喉中分别为3.7e2 pfu/ml和1.4 e3 pfu/ml，BAL中分别为2 e2 pfu/ml和3.7e2 pfu/ml，Gps 1和2分别为。这些结果证实了模型，并决定将10 e4 pfu/ml接种剂量用于随后的两项有效性研究。 进行了两项化合物连续静脉输注的疗效研究。 研究2有15只动物，分为三组，用GPS。研究1和2，治疗和预防组，TMC 353121以50 ng/ml的血浆水平给药，Gp 3为载体对照组;研究3将12只动物分为三组，Gp 1和2治疗组仅以5 ng/ml或500 ng/ml的血浆水平给药TMC 353121，Gp 3为载体对照组。 对动物进行预处理，使其穿上夹克和拴系系统，进行植入导管的手术，并开始输注。动物研究最近已经完成，但是，我们仍在进行噬菌斑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.