Generation of monoclonal and polyclonal antibodies to neolacto-series ganglioside

新乳系列神经节苷脂单克隆和多克隆抗体的生成

• 批准号: 7790685
• 负责人: AMJAD A. ILYAS
• 金额: $ 7.8万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7790685
• 关键词: Acute; Adult; Affinity; Animal Model; Animals; Antibodies; Antibody Affinity; Antigen Targeting; Antigens; Autoantibodies; Autoimmune Diseases; Autoimmune Process; Axonal Neuropathy; Biological Process; Blood; C3H/HeN Mouse; Canis familiaris; Carrier Proteins; Cattle; Cavia; Cell Line; Cell membrane; Cells; Development; Disease; Enzyme-Linked Immunosorbent Assay; Erythrocytes; Family; Felis catus; Female; Freund' s Adjuvant; Gangliosides; Generations; Glycolipids; Glycosphingolipids; Guillain-Barré Syndrome; Health; Human; Hybridomas; Immunization; Immunoglobulin G; Inbred BALB C Mice; Keyhole Limpet Hemocyanin; LM1 ganglioside; Laboratories; Lead; Link; Miller Fisher Syndrome; Monitor; Monoclonal Antibodies; Motor; Multiple Myeloma; Mus; Myelin; Nervous system structure; Neuraxis; Neuropathy; New Zealand; Oligosaccharides; Oryctolagus cuniculus; Peripheral Nerves; Peripheral Nervous System; Peripheral Nervous System Diseases; Peritoneal Fluid; Physiological; Plasma Cells; Play; Procedures; Rattus; Reagent; Role; Series; Signal Transduction; Specificity; Spleen; Structure; Variant; anti-IgG; base; in vivo; incomplete Freund' s adjuvant; polyclonal antibody; public health relevance; tool

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.

描述（由申请人提供）：神经节苷脂是存在于浆细胞膜中的唾液酸化鞘糖脂。它们特别集中在神经系统中。神经节苷脂被认为在细胞相互作用和信号转导中发挥着重要作用。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。虽然神经节系列神经节苷脂存在于中枢神经系统（CNS）和周围神经系统（PNS）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LM1和Hex-LM1。神经节苷脂被认为是多种自身免疫性神经病中的抗原。例如，抗 GM1 抗体与运动神经病相关，抗 GD1a 抗体与急性运动轴突神经病相关，抗 LM1 抗体与吉兰-巴利综合征 (GBS) 相关，抗 GQ1b 抗体与米勒·费希尔综合征（GBS 的一种变体）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。这部分是由于缺乏针对这些神经节苷脂的特异性探针。缺乏特异性抗体等探针的原因之一是很难从周围神经中分离出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。我们使用人类血液来分离和纯化大量的LM1。本研究的总体目标是开发针对新乳糖系列神经节苷脂的高亲和力特异性抗体。我们建议通过两个具体目标来实现我们的目标。在具体目标 1 中，我们将使用与匙孔血蓝蛋白 (KLH) 缀合的纯化 LM1 作为免疫原来开发小鼠单克隆抗体。在具体目标 2 中，我们将使用与 KLH 混合并乳化在弗氏佐剂中的 LM1 对新西兰白兔进行免疫，从而在新西兰白兔中产生多克隆抗体。我们的研究应该会导致针对 LM1 和 Hex-LM1 的高亲和力 IgG 抗体的开发。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它们在周围神经中的定位和生物学功能提供非常强大的工具。这些抗体对于描述其在周围神经系统自身免疫性疾病中的致病潜力也非常有用。公共健康相关性：目前尚无特定试剂，例如针对周围神经系统新乳糖系列神经节苷脂的单克隆抗体。开发针对这些重要神经节苷脂的高亲和力抗体将为探测它们在健康和疾病中的生理功能提供强大的工具。