Ab5 Toxins Nasal Adjuvant Target the Olfactory Bulbs/CNS

Ab5 毒素鼻佐剂靶向嗅球/中枢神经系统

• 批准号: 6711826
• 负责人: Kohtaro Fujihashi
• 金额: $ 25.11万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6711826
• 关键词: Escherichia coli; active immunization; antigen antibody reaction; apoptosis; astrocytes; bacterial toxins; bacterial vaccines; central nervous system; cholera vaccine; drug adverse effect; gangliosidosis GM1; in situ hybridization; inhalation drug administration; laboratory mouse; microarray technology; microglia; neuroimmunomodulation; neuropathology; neurotoxins; nonhuman therapy evaluation; olfactory lobe; protein binding; tissue /cell culture; toxicant interaction

DESCRIPTION: (Adapted from Applicant's Abstract) The intranasal route of vaccination has become the predominant one used m mucosal immunology in order to induce both systemic and mucosal T cell and antibody (Ab) responses. Nasal application of vaccines with adjuvants is easy to perform, requires l0 - 20 fold less material than does oral immunization and the vaccine and adjuvant are not subjected to degradative enzymes as they are in the GI tract. Despite these advantages, it is often not appreciated that the nasal region is extensively innervated by the olfactory bulb, which is directly connected via olfactory nerves to the olfactory epithelium. Both native enterotoxins like cholera and E. coli labile toxin (CT and LT), and nontoxic derivatives have pentameric B subunits, which bind to GM1 (CT) and to GM2 and asialo GM1 (LT) which are expressed by the olfactory epithelium and the associated olfactory nerves. It was shown that either CT or anti-GM1 Abs induced CNS lesions when injected into the lumbosacral subarachnoid space. This toxicity is consistent with expression of GM1 by astroglia and neurons of the CNS and by microglial cells of the cerebellar cortex. We have therefore postulated that intranasal application of nCT or nontoxic mCTs, which have intact (pentameric) CT-B, could enter the CNS through GM1 binding to olfactory epithelium and nerves with subsequent transport into the olfactory bulb. This would postulate that retrograde axonal transport would bypass the blood / brain barrier. Further, either nCT or mCT could potentially damage the CNS and even induce CNS uptake of co-nasally delivered vaccine proteins. The first Specific Aim to test this hypothesis will actually examine the trafficking of nCT or mCTs in olfactory epithelium and the CNS and whether these molecules induce neuropathologic changes. The second Specific Aim will determine if nCT or mCTs also induce uptake and trafficking of co-administered vaccine proteins into the nasal olfactory epithelium, nerves, and bulb. The third Specific Aim will determine potential toxicity of nCT and mCTs for cultured astrocytes and microglial cells, including inflammatory cytokine production. The fourth Specific Aim will extend this analysis of toxic, inflammatory and apoptotic responses to the nerve cells themselves in the olfactory bulb when exposed to nCT or mCTs. The final Specific Aim will assess long-term effects of CT-B binding to GM1 on neural cells, including the induction of autoimmune anti-GM1 Abs. These proposed studies will provide essential new information to determine potential side effects of CT-B derivatives as nasal adjuvants, and point to ways where these toxic effects can be avoided.

描述：（改编自申请人的摘要）鼻内路线 疫苗接种已成为主要的粘膜免疫学 诱导全身和粘膜T细胞和抗体（AB）反应。鼻 用佐剂应用疫苗很容易执行，需要L0-20 折叠比口服免疫的材料少，疫苗和佐剂是 不受胃肠道中的降解酶的影响。尽管如此 优势，通常不认为鼻部广泛 由嗅球支配，直接通过嗅觉连接 神经到嗅觉上皮。两种本地肠毒素，例如霍乱和 大肠杆菌不稳定毒素（CT和LT）和无毒衍生物具有五聚体B 与GM1（CT）和GM2和Asialo GM1（LT）结合的亚基 由嗅觉上皮和相关的嗅觉神经表达。它 显示出注射到 腰椎蛛网膜下腔空间。这种毒性与表达一致 Astroglia和CNS神经元的GM1和小胶质细胞的神经元 小脑皮质。因此，我们假设鼻内应用 具有完整（五聚体）CT-B的NCT或无毒MCT可以进入CNS 通过GM1结合与嗅觉上皮和神经的结合 运输到嗅球。这将假设逆行轴突 运输将绕过血液 /脑屏障。此外，无论是NCT还是MCT 可能会损害中枢神经系统，甚至诱导CNS的共同吸收 输送的疫苗蛋白。检验该假设的第一个特定目的将 实际检查NCT或MCT在嗅觉上皮和 中枢神经系统以及这些分子是否诱导神经病理学变化。第二个 具体目标将确定NCT或MCT是否也引起吸收和贩运 共同采用的疫苗蛋白进入鼻嗅觉上皮， 神经和灯泡。第三个特定目的将确定潜在的毒性 NCT和MCT用于培养的星形胶质细胞和小胶质细胞，包括 炎症性细胞因子产生。第四个特定目标将扩展到这一点 分析对神经细胞的有毒，炎症和凋亡反应 暴露于NCT或MCT时，自己在嗅球中。决赛 具体目标将评估CT-B与GM1的长期影响对神经的结合 细胞，包括诱导自身免疫性抗GM1 ABS。这些提议 研究将提供重要的新信息以确定潜在的一面 CT-B衍生物作为鼻佐剂的影响，并指出这些方法 可以避免有毒作用。