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转染表达功能性重组细胞间黏附分子 1(ICAM-1)的 HEK293 细胞--与严重恶性疟原虫疟疾相关的一种受体。

Transfected HEK293 cells expressing functional recombinant intercellular adhesion molecule 1 (ICAM-1)--a receptor associated with severe Plasmodium falciparum malaria.

机构信息

Centre for Medical Parasitology at the Department of International Health, Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen and at Department of Infectious Diseases, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark.

出版信息

PLoS One. 2013 Jul 25;8(7):e69999. doi: 10.1371/journal.pone.0069999. Print 2013.

DOI:10.1371/journal.pone.0069999
PMID:23936131
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3723725/
Abstract

Intercellular adhesion molecule 1 (ICAM-1) is a membrane-bound glycoprotein expressed on endothelial cells and cells of the immune system. Human ICAM-1 mediates adhesion and migration of leucocytes, and is implicated in inflammatory pathologies, autoimmune diseases and in many cancer processes. Additionally, ICAM-1 acts as receptor for pathogens like human rhinovirus and Plasmodium falciparum malaria parasites. A group of related P. falciparum erythrocyte membrane protein 1 (PfEMP1) domains, the DBLβ, mediates ICAM-1 binding of P. falciparum-infected erythrocytes. This ICAM‑1-binding phenotype has been suggested to be involved in the development of cerebral malaria. However, more studies identifying cross-reactive antibody and ICAM-1-binding epitopes and the establishment of a clinical link between DBLβ expression and e.g. cerebral malaria are needed before the DBLβ domains can be put forward as vaccine candidates and go into clinical trials. Such studies require availability of functional recombinant ICAM-1 in large quantities. In this study, we compared recombinant ICAM-1 expressed in HEK293 and COS-7 cells with mouse myeloma NS0 ICAM-1 purchased from a commercial vendor in terms of protein purity, yield, fold, ability to bind DBLβ, and relative cost. We present a HEK293 cell-based, high-yield expression and purification scheme for producing inexpensive, functional ICAM‑1. ICAM-1 expressed in HEK293 is applicable to malaria research and can also be useful in other research fields.

摘要

细胞间黏附分子 1(ICAM-1)是一种表达在血管内皮细胞和免疫系统细胞上的膜结合糖蛋白。人类 ICAM-1 介导白细胞的黏附和迁移,并与炎症性病理、自身免疫性疾病和许多癌症过程有关。此外,ICAM-1 作为人类鼻病毒和恶性疟原虫疟原虫等病原体的受体。一组相关的恶性疟原虫红细胞膜蛋白 1(PfEMP1)结构域,即 DBLβ,介导 PfEMP1 感染的红细胞与 ICAM-1 的结合。这种与 ICAM-1 结合的表型被认为与脑型疟疾的发展有关。然而,在 DBLβ 结构域可以作为疫苗候选物并进入临床试验之前,还需要更多的研究来确定交叉反应性抗体和与 ICAM-1 结合的表位,并建立 DBLβ 表达与例如脑型疟疾之间的临床联系。这些研究需要大量功能性重组 ICAM-1。在这项研究中,我们比较了在 HEK293 和 COS-7 细胞中表达的重组 ICAM-1 与从商业供应商购买的鼠骨髓瘤 NS0 ICAM-1 的蛋白纯度、产量、折叠、与 DBLβ 结合的能力以及相对成本。我们提出了一种基于 HEK293 细胞的高效表达和纯化方案,用于生产廉价、功能性的 ICAM-1。在 HEK293 中表达的 ICAM-1 适用于疟疾研究,也可用于其他研究领域。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddc7/3723725/8d92f523885a/pone.0069999.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddc7/3723725/4b2e9b705726/pone.0069999.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddc7/3723725/65167c4dfca0/pone.0069999.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddc7/3723725/ec975ee9340c/pone.0069999.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddc7/3723725/8d92f523885a/pone.0069999.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddc7/3723725/4b2e9b705726/pone.0069999.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddc7/3723725/65167c4dfca0/pone.0069999.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddc7/3723725/ec975ee9340c/pone.0069999.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddc7/3723725/8d92f523885a/pone.0069999.g004.jpg

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