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: 6634552
• 负责人: Jerry R McGhee
• 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 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6634552
• 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）鼻内途径 疫苗接种已成为粘膜免疫学中使用的主要方法 诱导全身和粘膜 T 细胞和抗体 (Ab) 反应。鼻腔 疫苗与佐剂的应用很容易进行，需要 l0 - 20 比口服免疫折叠更少的材料，并且疫苗和佐剂 由于它们在胃肠道中，因此不会受到降解酶的影响。尽管有这些 优点，但人们常常没有意识到鼻部区域广泛 受嗅球支配，嗅球通过嗅觉直接连接 神经连接到嗅上皮。天然肠毒素如霍乱和 大肠杆菌不稳定毒素（CT 和 LT），无毒衍生物具有五聚体 B 亚基，与 GM1 (CT) 以及 GM2 和 asialo GM1 (LT) 结合， 由嗅上皮和相关的嗅神经表达。它 结果表明，CT 或抗 GM1 Abs 注射到体内时会引起 CNS 损伤。 腰骶部蛛网膜下腔。这种毒性与表达是一致的 GM1 通过中枢神经系统的星形胶质细胞和神经元以及小神经胶质细胞 小脑皮质。因此，我们假设鼻内应用 nCT 或无毒 mCT 具有完整（五聚体）CT-B，可以进入中枢神经系统 通过 GM1 与嗅觉上皮和神经结合，随后 运输到嗅球。这假设逆行轴突 运输将绕过血/脑屏障。此外，nCT 或 mCT 可能会损害中枢神经系统，甚至诱导中枢神经系统吸收鼻腔药物 递送疫苗蛋白。检验该假设的第一个具体目标将是 实际检查 nCT 或 mCT 在嗅觉上皮中的贩运以及 中枢神经系统以及这些分子是否会引起神经病理学变化。第二个 具体目标将确定 nCT 或 mCT 是否也会诱导摄取和贩运 共同施用的疫苗蛋白进入鼻嗅上皮， 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 炎症细胞因子的产生。第四个具体目标将扩展这一目标 分析神经细胞的毒性、炎症和凋亡反应 当暴露于 nCT 或 mCT 时，它们本身位于嗅球中。决赛 具体目标将评估 CT-B 与 GM1 结合对神经的长期影响 细胞，包括诱导自身免疫性抗 GM1 抗体。这些建议 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作用，并指出这些作用的方法 可以避免毒性作用。