Ab5 Toxins Nasal Adjuvant Target the Olfactory Bulbs/CNS

描述：(改编自申请者摘要)鼻腔途径 疫苗接种已成为黏膜免疫学的主要方法。 诱导全身和粘膜T细胞及抗体(Ab)反应。鼻腔 应用带有佐剂的疫苗很容易操作，需要10-20个 与口服免疫相比，折叠材料更少，疫苗和佐剂 不像在胃肠道中那样受到降解酶的影响。尽管如此 优势，但人们往往没有意识到鼻区广泛 由嗅球支配，通过嗅觉直接连接 神经传导至嗅觉上皮。既有霍乱等天然肠毒素，也有 大肠杆菌不稳定毒素(CT和LT)及其无毒衍生物具有五聚体B 与GM1(CT)、GM2和asialo GM1(LT)结合的亚基，它们是 由嗅觉上皮和相关的嗅神经表达。它 结果表明，无论是CT抗体还是抗GM1抗体，在注射到 腰椎蛛网膜下腔。这种毒性与表现出来的 星形胶质细胞和中枢神经系统神经元及小胶质细胞表达GM1 小脑皮层。因此，我们假设鼻腔内应用 具有完整(五聚体)CT-B的NCT或无毒MCT可进入CNS 通过GM1与嗅觉上皮和神经结合，从而 输送到嗅球中。这将假设逆行轴突 交通工具将绕过血/脑屏障。此外，无论是NCT还是MCT 可能会潜在地损害中枢神经系统，甚至诱导鼻咽癌对中枢神经系统的摄取 运送疫苗蛋白质。检验这一假设的第一个具体目标是 实际检查NCT或MCT在嗅上皮细胞中的运输 中枢神经系统，以及这些分子是否引起神经病理变化。第二 具体目标将决定NCT或MCT是否也会导致摄取和贩运 联合注射疫苗蛋白进入鼻嗅觉上皮， 神经和球茎。第三个具体目标将确定 培养的星形胶质细胞和小胶质细胞的NCT和MCT，包括 炎性细胞因子的产生。第四个具体目标将扩展这一点 神经细胞的毒性、炎症和凋亡反应分析 当接触NCT或MCT时，他们自己在嗅球中。决赛 特定目的将评估CT-B与GM1结合对神经的长期影响 细胞，包括诱导自身免疫的抗GM1抗体。这些建议 研究将提供重要的新信息，以确定潜在的方面 CT-B衍生物作为鼻佐剂的作用，并指出了这些 毒性效应是可以避免的。

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.