Antibody Generation of ROS and Macular Degeneration

ROS 和黄斑变性的抗体生成

• 批准号: 7096710
• 负责人: PAUL WENTWORTH
• 金额: $ 27.89万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7096710
• 关键词: antibody; culture; human; immunoglobulins; lighting; macular degeneration; oxidative stress; singlet oxygen

DESCRIPTION (provided by applicant): Age-related macular degeneration (AMD) is the leading cause of blind registration in the developing world and yet its pathogenesis remains poorly understood. However, there is increasing evidence to support a role for the immune system and oxidative stress, arising from an imbalance in the production and destruction of reactive oxygen species (ROS) such as superoxide anion, hydrogen peroxide and singlet oxygen, in disease progression. There is also clear evidence that photochemical sensitization of chromophores stored within lipofuscin in the retinal pigment epithelial (RPE) cells via absorption of both ultraviolet and visible light, leads to ROS generation and damage in the ageing eye. We have recently shown that all antibody molecules, regardless of species or antigenic determinant, have an intrinsic ability to generate a cascade of potent oxidants when presented with either a chemical or photochemical source of singlet dioxygen. The end product of the antibody-catalyzed water-oxidation pathway (ACWOP) is hydrogen peroxide and thus the implication for this pathway to play a role in AMD via interception of photochemically generated singlet oxygen is a real possibility and would lead to an increase in overall oxidative stress. We intend to investigate this hypothesis with the three following specific aims: 1. Specific Aim #1 To determine whether oxidants generated by the ACWOP, are damaging to passaged human RPE cells or primary cultured human RPE cells, in terms of cytotoxicity, apoptosis and expression of heat shock protein (Hsp) 27. 2. Specific Aim #2 To determine whether the ACWOP can be activated by photoirradiation of A2E-loaded RPE cells and whether such activation enhances photochemical damage to these passaged and primary cultures of human RPE cells. 3. Specific Aim #3 To determine whether the ACWOP when activated in whole blood, plasma and serum can affect RPE cell survival

描述（由申请人提供）：视网膜相关性黄斑变性（AMD）是发展中国家失明登记的主要原因，但其发病机制仍知之甚少。然而，越来越多的证据支持免疫系统和氧化应激在疾病进展中的作用，氧化应激是由活性氧（ROS）（如超氧阴离子、过氧化氢和单线态氧）的产生和破坏不平衡引起的。还有明确的证据表明，通过吸收紫外线和可见光，储存在视网膜色素上皮（RPE）细胞中脂褐质内的发色团的光化学敏化导致ROS产生和老化眼睛的损伤。我们最近已经表明，所有的抗体分子，无论物种或抗原决定簇，有一个内在的能力，以产生一个强大的氧化剂级联时，无论是化学或光化学源的单线态分子氧。抗体催化的水氧化途径（ACWOP）的终产物是过氧化氢，因此暗示该途径通过光化学产生的单线态氧的拦截在AMD中发挥作用是真实的可能性，并且将导致总体氧化应激的增加。我们打算调查这一假设与以下三个具体目标：1。具体目的#1确定ACWOP产生的氧化剂是否在细胞毒性、细胞凋亡和热休克蛋白（Hsp）27的表达方面对传代的人RPE细胞或原代培养的人RPE细胞造成损害。2.具体目的#2确定ACWOP是否可以通过光照射负载A2E的RPE细胞来激活，以及这种激活是否增强了对人RPE细胞的这些传代和原代培养物的光化学损伤。3.具体目的#3确定ACWOP在全血、血浆和血清中活化时是否可影响RPE细胞存活