GENE THERAPY OF BRAIN TUMORS

这份分包合同调查的是行政和 新型病毒载体(逆转录病毒、单纯疱疹病毒)的神经致病作用 类型1和疱疹病毒扩增)在非人灵长类动物。最初，我们使用 检查恒河猴疱疹病毒载体的渗透情况 脑内注射后(使用一种新的立体定向 微量注射装置)和宿主对该载体的响应。至 我们已经检查了两只动物，这两只动物都被给予了一般 麻醉，放置在立体定向器中，以及开颅手术 在无菌条件下进行的。使用微量注射装置 插入了四根彼此相距5 mm的26号针头 右额叶皮质有10毫米深。疱疹病毒(共1×109 PFU在100ul/次注射)超过120分钟。之后 拔除注射、针头和立体定向器， 开颅手术部位通过常规手术程序关闭。一 动物在注射后48小时和14小时后被安乐死并进行检查 注射后数天。两只动物都是完全尸检 考试。两只动物都有局限性的广泛性软化 注射。在炎性细胞中检测到病毒蛋白 染毒48小时后动物偶见神经元被安乐死 HSV-1突变株，但在14岁时安乐死和检查的动物中不存在 注射后数天。我们的初步研究表明，天真的 恒河猴(恒河猴疱疹病毒B阴性)可感染 HSV-1，并对HSV-1产生抗体反应，但不发展 泛化。虽然恒河猴是一个合适的模型 某些HSV-1研究，以充分评估HSV-1的安全性 在人类中，我们目前正在使用猫头猴(Aotus sp.)，这是一种 容易感染全身性HSV-1。目前，两只猫头鹰 猴子已感染HSV-1变异株(108 Pfu) 大脑内(如上所述)和实施安乐死并在 感染后4周和12周。无论是病毒蛋白还是 在中枢神经系统和其他组织中检测到显著的组织病理学变化 组织检查。年内将对更多的动物进行调查 这项提案中的三个。

This subcontract investigates the administration and neuropathogenesis of novel virus vectors (retrovirus, herpes simplex type 1 and herpes-amplicon) in nonhuman primates. Initially, we used rhesus monkeys to examine the infiltration of a herpesvirus vector after intracerebral injection (using a novel stereotactic micro-injection apparatus) and the host response to this vector. To date we have examined two animals both animals were given general anesthesia, placed in a sterotactic apparatus, and craniotomies performed under sterile conditions. Using a micro-injection apparatus four 26 gauge needles placed 5 mm apart from one another were inserted 10 mm into the right frontal cortex. Herpesvirus (a total of 1 X 109 pfu in 100 ul/ injection) were injected over 120 minutes. After injection, needles and stereotactic apparatus were removed and the craniotomy sites were closed by routine surgical procedures. One animal was euthanized and examined 48 hours post-injection and one 14 days post-injection. Both animals had complete post-mortem examinations. Both animals had focally extensive malacia at the site of injection. Viral protein was detected in inflammatory cells and occasional neurons in the animal euthanized 48 hours after exposure to the HSV-1 mutant, but not in the animal euthanized and examined at 14 days post injection. Our preliminary studies have shown that naive rhesus monkeys (rhesus herpesvirus B negative) can be infected with HSV-1 and mount an antibody response to HSV-1, but do not develop generalized. Although the rhesus monkey is a suitable model for certain HSV-1 studies, to adequately assess the safety of HSV-1 mutants in man, we are currently using owl monkeys (Aotus sp.), which are susceptible to generalized HSV-1 infection. Presently, two owl monkeys have been infected with HSV-1 mutants (108 pfu) intracerebrally (as described above) and euthanized and examined at four and twelve weeks post infection. Neither viral protein nor significant histopathologic changes were detected in CNS and other tissues examined. Additional animals will be investigated in year three of this proposal.