Cargando…

Transfected HEK293 Cells Expressing Functional Recombinant Intercellular Adhesion Molecule 1 (ICAM-1) – A Receptor Associated with Severe Plasmodium falciparum Malaria

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. Addi...

Descripción completa

Detalles Bibliográficos
Autores principales: Bengtsson, Anja, Joergensen, Louise, Barbati, Zachary R., Craig, Alister, Hviid, Lars, Jensen, Anja T. R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3723725/
https://www.ncbi.nlm.nih.gov/pubmed/23936131
http://dx.doi.org/10.1371/journal.pone.0069999
_version_ 1782278322711953408
author Bengtsson, Anja
Joergensen, Louise
Barbati, Zachary R.
Craig, Alister
Hviid, Lars
Jensen, Anja T. R.
author_facet Bengtsson, Anja
Joergensen, Louise
Barbati, Zachary R.
Craig, Alister
Hviid, Lars
Jensen, Anja T. R.
author_sort Bengtsson, Anja
collection PubMed
description 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.
format Online
Article
Text
id pubmed-3723725
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-37237252013-08-09 Transfected HEK293 Cells Expressing Functional Recombinant Intercellular Adhesion Molecule 1 (ICAM-1) – A Receptor Associated with Severe Plasmodium falciparum Malaria Bengtsson, Anja Joergensen, Louise Barbati, Zachary R. Craig, Alister Hviid, Lars Jensen, Anja T. R. PLoS One Research Article 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. Public Library of Science 2013-07-25 /pmc/articles/PMC3723725/ /pubmed/23936131 http://dx.doi.org/10.1371/journal.pone.0069999 Text en © 2013 Bengtsson et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Bengtsson, Anja
Joergensen, Louise
Barbati, Zachary R.
Craig, Alister
Hviid, Lars
Jensen, Anja T. R.
Transfected HEK293 Cells Expressing Functional Recombinant Intercellular Adhesion Molecule 1 (ICAM-1) – A Receptor Associated with Severe Plasmodium falciparum Malaria
title Transfected HEK293 Cells Expressing Functional Recombinant Intercellular Adhesion Molecule 1 (ICAM-1) – A Receptor Associated with Severe Plasmodium falciparum Malaria
title_full Transfected HEK293 Cells Expressing Functional Recombinant Intercellular Adhesion Molecule 1 (ICAM-1) – A Receptor Associated with Severe Plasmodium falciparum Malaria
title_fullStr Transfected HEK293 Cells Expressing Functional Recombinant Intercellular Adhesion Molecule 1 (ICAM-1) – A Receptor Associated with Severe Plasmodium falciparum Malaria
title_full_unstemmed Transfected HEK293 Cells Expressing Functional Recombinant Intercellular Adhesion Molecule 1 (ICAM-1) – A Receptor Associated with Severe Plasmodium falciparum Malaria
title_short Transfected HEK293 Cells Expressing Functional Recombinant Intercellular Adhesion Molecule 1 (ICAM-1) – A Receptor Associated with Severe Plasmodium falciparum Malaria
title_sort transfected hek293 cells expressing functional recombinant intercellular adhesion molecule 1 (icam-1) – a receptor associated with severe plasmodium falciparum malaria
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3723725/
https://www.ncbi.nlm.nih.gov/pubmed/23936131
http://dx.doi.org/10.1371/journal.pone.0069999
work_keys_str_mv AT bengtssonanja transfectedhek293cellsexpressingfunctionalrecombinantintercellularadhesionmolecule1icam1areceptorassociatedwithsevereplasmodiumfalciparummalaria
AT joergensenlouise transfectedhek293cellsexpressingfunctionalrecombinantintercellularadhesionmolecule1icam1areceptorassociatedwithsevereplasmodiumfalciparummalaria
AT barbatizacharyr transfectedhek293cellsexpressingfunctionalrecombinantintercellularadhesionmolecule1icam1areceptorassociatedwithsevereplasmodiumfalciparummalaria
AT craigalister transfectedhek293cellsexpressingfunctionalrecombinantintercellularadhesionmolecule1icam1areceptorassociatedwithsevereplasmodiumfalciparummalaria
AT hviidlars transfectedhek293cellsexpressingfunctionalrecombinantintercellularadhesionmolecule1icam1areceptorassociatedwithsevereplasmodiumfalciparummalaria
AT jensenanjatr transfectedhek293cellsexpressingfunctionalrecombinantintercellularadhesionmolecule1icam1areceptorassociatedwithsevereplasmodiumfalciparummalaria