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: 6321437
• 负责人: Jerry R McGhee
• 金额: $ 27.83万
• 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 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6321437
• 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） 疫苗接种已成为粘膜免疫学中使用的主要方法， 以诱导全身和粘膜T细胞和抗体（Ab）应答。鼻 使用含有佐剂的疫苗很容易，需要10 - 20 比口服免疫少1倍的材料，并且疫苗和佐剂是 不像在胃肠道中那样受到降解酶的作用。尽管有这些 然而，尽管鼻区域具有许多优点，但通常不认识到鼻区域广泛地被 由嗅球支配，嗅球通过嗅觉直接连接 神经到嗅上皮。霍乱等天然肠毒素， E.大肠杆菌不稳定毒素（CT和LT）和无毒衍生物具有五聚体B 亚基，其结合GM 1（CT）和GM 2和脱唾液酸GM 1（LT）， 由嗅上皮和相关的嗅神经表达。它 结果表明，当注射到中枢神经系统中时，CT或抗GM 1抗体均诱导CNS病变， 腰骶部蛛网膜下腔这种毒性与表达一致 GM 1的星形胶质细胞和神经元的中枢神经系统和小胶质细胞的 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可进入CNS 通过GM 1与嗅上皮和神经的结合， 运输到嗅球。这就假定逆行轴突 运输会绕过血脑屏障此外，nCT或mCT 可能潜在地损害CNS，甚至诱导CNS摄取鼻内 递送疫苗蛋白质。检验这一假设的第一个具体目标将 实际上检查了嗅上皮中nCT或mCT的运输， CNS以及这些分子是否诱导神经病理学变化。第二 特定目的将确定nCT或mCT是否也诱导摄取和转运 鼻嗅上皮中的疫苗蛋白质， 神经和灯泡。第三个具体目标将确定潜在的毒性 用于培养的星形胶质细胞和小胶质细胞的nCT和mCT，包括 炎性细胞因子产生。第四个具体目标将扩大这一点， 对神经细胞的毒性、炎症和凋亡反应的分析 当暴露于nCT或mCT时，它们自身在嗅球中。最终 Specific Aim将评估CT-B与GM 1结合对神经细胞的长期影响。 细胞，包括诱导自身免疫性抗GM 1抗体。这些拟议 研究将提供重要的新信息，以确定潜在的 CT-B衍生物作为鼻佐剂的作用，并指出这些方法 可以避免毒性作用。