Development of Robust Culture Systems for the Human Splenic Littoral Cell

人脾滨细胞稳健培养系统的开发

• 批准号: 8428539
• 负责人: JOYCE DIANE FINGEROTH
• 金额: $ 8.7万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-15 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8428539
• 关键词: Antigen-Antibody Complex; Antigens; Apoptotic; Appearance; Autoimmune Diseases; Blood; Blood Circulation; Blood Platelets; Blood Vessels; Blood flow; Bypass; CD8 Antigens; Cell Culture System; Cell Line; Cell Nucleus; Cell physiology; Cells; Cellular biology; Communicable Diseases; Complex; Cytoplasm; Cytoplasmic Filaments; Deposition; Development; Discontinuous Capillary; Disease; Distant; Endothelial Cells; Evolution; Filtration; Foundations; Future; Gene Proteins; Growth; Heart Diseases; Hematopoietic; Hemoglobinopathies; Hemolytic Anemia; Hemosiderin; Human; Immune; Immune system; Immunologics; Immunologist; Immunology; In Situ; Individual; Infection; Infectious Agent; Investigation; Laboratories; Lead; Left; Leukocytes; Lymphatic System; Lysosomes; Malaria; Malignant Neoplasms; Malpighian corpuscles; Mammals; Mediator of activation protein; Methods; Mus; Necrosis; Organ; Pathologist; Pathology; Persons; Phagocytes; Phagocytosis; Population; Primates; Procedures; Process; Proteins; RNA; Regulation; Research Methodology; Risk; Sepsis; Sinus; Site; Smooth Muscle; Spleen; Splenic Red Pulp; Splenocyte; Stress Fibers; Structure; System; Therapeutic; Ursidae Family; Venous; Vesicle; Work; base; lymph nodes; macrophage; man; monocyte; mortality; neoplastic cell; novel; pathogen; preclinical study; prevent; protein expression; public health relevance; repaired; success; therapeutic development; trafficking; venous sinus

DESCRIPTION (provided by applicant): The spleen is the largest secondary immune system organ in man. Individuals who are asplenic die at an increased rate from overwhelming sepsis. They are also at greater risk of autoimmune disease, some cancers and ischemic cardiac disease. Enhanced mortality is attributed to lack of phagocytes sequestered in spleen that efficiently eliminate appropriate targets, though related cells can be found at other sites. Although the overall organization of splenic white pulp is similar to that of lymph nodes, spleen is not connected to the lymphatic system. Instead, pathogens, RBCs, apoptotic/necrotic, altered and tumor cells all are delivered to and leave the spleen in blood. In contrast to white pulp, splenic red pulp serves as both a reservoir and a filter that determines whether cells coursing therein are retained, destroyed or returned to the circulation. The human venous sinus-lining cell, known as the littoral cell (LC), comprises ~30% of red pulp. Increasing evidence indicates these highly specialized splenocytes are the major determinant of whether RBCs and others are destroyed, retained or returned to the circulation, though the mechanisms that regulate their function are entirely unknown. Others and we showed that LCs are not a classical endothelial cells as they bear multiple macrophage and smooth muscle-associated antigens, are CD8+ and lack traditional markers of endothelial and also hematopoietic lineage. So, why is so little known about LC function? The acquisition, purification and culture of LCs have historically proved very difficult. Based on our recent success in obtaining fresh spleen as well as identifying markers that distinguish LCs from other splenocytes, we are now prepared to further isolate and profile LC gene/protein expression (AIM I). Although cells such as LCs that form specialized structures within complex organs are often hard to propagate ex vivo, overcoming these limitations would represent a significant advance in understanding LC function(s). We have assembled a group of expert endothelial cell biologist-immunologists, and pathologists who will provide us with in depth guidance as we develop diverse LC culture systems (AIM II) that are both physiologically relevant and tractable. These methods will enable discovery of the mechanisms that underlie LC function and lead to development of therapeutics able to modulate splenic filtration in different disease states (e.g. sepsis, malaria, hemoglobinopathies and others).

描述（由申请人提供）：脾脏是人类中最大的二级免疫系统器官。无生气的人死于压倒性败血症的速度。它们还具有更大的自身免疫性疾病，一些癌症和缺血性心脏病的风险。增强的死亡率归因于脾脏中缺乏封存的吞噬细胞，从而有效地消除了适当的靶标，尽管可以在其他部位找到相关的细胞。尽管脾白浆的总体组织与淋巴结相似，但脾脏与淋巴系统无关。取而代之的是，病原体，rbcs，凋亡/坏死，改变和肿瘤细胞均被递送到血液中并留在血液中。与白色果肉相反，脾红色纸浆既是储层，又是确定在其中的细胞保留，破坏或返回到循环的过滤器。人静脉窦细胞，称为沿海细胞（LC），占红浆的约30％。越来越多的证据表明，这些高度专业的脾细胞是RBC和其他人是否被破坏，保留或返回到循环的主要决定因素，尽管调节其功能的机制是完全未知的。其他，我们表明LCS不是经典的内皮细胞，因为它们具有多个巨噬细胞和平滑肌相关的抗原，是CD8+，并且缺乏内皮和造血谱系的传统标记。那么，为什么对LC功能鲜为人知？历史上，LC的获取，净化和文化非常困难。基于我们最近在获得新鲜脾的成功以及鉴定将LC与其他脾细胞区分开的标记，我们现在准备进一步隔离和剖面LC基因/蛋白质表达（AIM I）。尽管在复杂器官中形成专门结构的LC等细胞通常很难传播离体，但克服这些局限性将代表理解LC功能的重大进步。我们已经组建了一组专家内皮细胞生物学家免疫学家和病理学家，他们将为我们提供深入的指导，因为我们开发了不同的LC培养系统（AIM II），这些系统在生理上相关且可拖延。这些方法将使能够发现LC功能基础的机制，并导致能够调节不同疾病状态下脾脏过滤的治疗剂（例如脓毒症，疟疾，血红蛋白疾病等）。