Ioannis Paspaltsis, Evangelia Kesidou, Olga Touloumi, Roza Lagoudaki, Marina Boziki, Martina Samiotaki, Dimitra Dafou, Theodoros Sklaviadis, Nikolaos Grigoriadis
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引用次数: 0
摘要
背景:神经前体细胞(NPC)的发现以及与之相关的深入研究为再生医学提供了新的方法,使其能够解决神经退行性疾病等问题。如今,NPCs 移植被认为是治疗这些疾病的最前沿方法,许多相关的临床试验已经完成或仍在进行中。然而,人们对鼻咽癌细胞的抗原性知之甚少,大多数研究都在探讨鼻咽癌细胞的抗原性是否会导致移植细胞的排斥反应:在这项研究中,我们按照标准免疫方案,对野生型 C57BL/6 小鼠皮下注射合成 NPCs 乳液后的抗原潜力进行了调查。免疫时表达的全部 IgG 重排被克隆到 Fab 噬菌体展示载体中。用生物平移技术从创建的噬菌体展示文库中筛选出与鼻咽癌裂解物蛋白相互作用的 Fab 表达克隆。克隆 65 的 IgG Fab 片段在不同的免疫测定中被证明对来自鼻咽癌裂解液和/或全脑裂解液的抗原有反应:结论:使用标准免疫方案注射鼻咽癌抗原,并应用Fab片段噬菌体展示技术,我们能够分离出至少一种单克隆IgG Fab片段,它能与不同的小鼠脑蛋白相互作用。目前还不清楚移植人大脑后宿主生物体内是否会产生这种抗体。
Application of antibody phage display to identify potential antigenic neural precursor cell proteins.
Background: The discovery of neural precursor cells (NPCs) and the concomitant intensive research in the field offer regenerative medicine novel approaches, enabling it to tackle conditions, such as neurodegenerative diseases. Transplantation of NPCs is nowadays considered a cutting-edge treatment for these conditions and many related clinical trials have been already completed or are still ongoing. However, little is known about the antigenicity of NPCs, with most studies addressing the question whether their antigenicity could lead to rejection of the transplanted cells.
Results: In this study we investigated the antigenic potential of syngeneic NPCs emulsion, upon subcutaneous (s.c.) administration to wild type C57BL/6 mice, following a standard immunization protocol. The whole IgG repertoire expressed upon immunization was cloned into a Fab phage display vector. From the created phage display library, Fab expressing clones interacting with NPCs lysate proteins were selected with the biopanning technique. The IgG Fab fragment from clone 65 proved to be reactive against antigens originating from NPCs lysates and/or whole brain lysate in diverse immunological assays.
Conclusions: Using a standard immunization protocol to administer NPCs antigens, and applying the Fab fragment phage display technique, we were able to isolate at least a monoclonal IgG Fab fragment, which interacts with different mouse brain proteins. It is not clear whether such antibodies are produced in the host organisms, following NPCs transplantation.
期刊介绍:
Journal of Biological Research-Thessaloniki is a peer-reviewed, open access, international journal that publishes articles providing novel insights into the major fields of biology.
Topics covered in Journal of Biological Research-Thessaloniki include, but are not limited to: molecular biology, cytology, genetics, evolutionary biology, morphology, development and differentiation, taxonomy, bioinformatics, physiology, marine biology, behaviour, ecology and conservation.