Generation of monoclonal and polyclonal antibodies to neolacto-series ganglioside

性状（由申请方提供）：神经节苷脂是存在于浆细胞膜中的唾液酸化鞘糖脂。尤其集中在神经系统。神经节苷脂被认为在细胞相互作用和信号转导中发挥重要作用。根据寡糖结构，神经节苷脂可分为几个主要家族，包括神经节苷脂系列和新乳糖系列。虽然神经节苷脂系列存在于中枢神经系统（CNS）和外周神经系统（PNS）两者中，但新乳糖系列神经节苷脂主要存在于PNS中。人PNS的主要新乳糖系列神经节苷脂是LM 1和Hex-LM 1。神经节苷脂作为抗原参与各种自身免疫性神经病。例如，抗GM1抗体与运动神经病相关，抗GD1a抗体与急性运动轴索神经病相关，抗LM1抗体与格林-巴利综合征（GBS）相关，抗GQ1b抗体与GBS变体米勒费舍尔综合征相关。新乳糖系列神经节苷脂抗体在自身免疫性周围神经病中的作用以及这些神经节苷脂的定位和功能仍然未知。这部分是由于缺乏这些神经节苷脂的特异性探针。缺乏探针如特异性抗体的一个原因是难以从外周神经中分离大量的这些神经节苷脂。有趣的是，LM1神经节苷脂也存在于人类红细胞中。我们已经使用人血分离和纯化了大量的LM1。本研究的总体目标是开发针对新乳系列神经节苷脂的高亲和力特异性抗体。我们建议在两个具体目标中实现我们的目标。在特定目标1中，我们将使用与钥孔血蓝蛋白（KLH）结合的纯化LM 1作为免疫原开发小鼠单克隆抗体。在特异性目标2中，我们将通过用与KLH混合并在弗氏佐剂中乳化的LM 1免疫新西兰白色兔来产生多克隆抗体。我们的研究将导致开发针对LM1和Hex-LM1的高亲和力IgG抗体。新乳糖系列神经节苷脂的多克隆抗体和一组单克隆抗体的可用性将为阐明其在周围神经中的定位和生物学功能提供非常强大的工具。这些抗体也将是非常有用的，以描绘其在周围神经系统的自身免疫性疾病的致病潜力。公共卫生相关性：特异性试剂如外周神经系统新乳糖系列神经节苷脂的单克隆抗体目前尚不可用。开发这些重要神经节苷脂的高亲和力抗体将为探索其在健康和疾病中的生理功能提供强有力的工具。

DESCRIPTION (provided by applicant): Gangliosides are sialylated glycosphingolipids present in plasma cell membranes. They are particularly concentrated in the nervous system. Gangliosides have been implicated as playing major roles in cellular interactions and signal transduction. Based on the oligosaccharide structures, gangliosides can be categorized into several major families, including ganglio-series and neolacto-series. While ganglio- series gangliosides are present in both the central nervous system (CNS) and peripheral nervous system (PNS), the neolacto-series gangliosides are present mainly in the PNS. The major neolacto- series gangliosides of human PNS are LM1 and Hex-LM1. Gangliosides have been implicated as antigens in various autoimmune neuropathies. For example, anti-GM1 antibodies are associated with motor neuropathy, anti-GD1a antibodies are associated with acute motor axonal neuropathy, anti-LM1 antibodies are associated with the Guillain-Barre syndrome (GBS) and anti-GQ1b antibodies are associated with Miller Fisher syndrome, a variant of GBS. The role of antibodies to neolacto-series gangliosides in autoimmune peripheral neuropathies and the localization and function of these gangliosides remain unknown. This is partly due to the lack of specific probes for these gangliosides. One reason for the lack of probes such as specific antibodies is that it is difficult to isolate large quantities of these gangliosides from peripheral nerves. Interestingly, LM1 ganglioside is also present in human red blood cells. We have used human blood to isolate and purify large quantities of LM1. The overall objective of this study is to develop high-affinity specific antibodies against neolacto-series gangliosides. We propose to achieve our objective in two specific aims. In Specific Aim 1, we will develop mouse monoclonal antibodies using purified LM1 conjugated to keyhole limpet hemocyanin (KLH) as an immunogen. In Specific Aim 2, we will generate polyclonal antibodies in New Zealand white rabbits by immunizing them with LM1 mixed with KLH and emulsified in Freund's adjuvant. Our study should lead to the development of high-affinity IgG antibodies to LM1 and Hex-LM1. The availability of polyclonal and a panel of monoclonal antibodies to the neolacto-series gangliosides will provide very powerful tools to elucidate their localization and biological function in the peripheral nerves. These antibodies will also be very useful to delineate their pathogenic potential in autoimmune diseases of the peripheral nervous system. PUBLIC HEALTH RELEVANCE: Specific reagents such as monoclonal antibodies to peripheral nervous system neolacto- series gangliosides are currently not available. Development of high-affinity antibodies to these important gangliosides would provide powerful tools to probe their physiological functions in health and disease.