isolation-human-umbilical-vein-endothelial-cells-their-use-study

Isolation of Human Umbilical Vein Endothelial Cells and Their Use in the Study of Neutrophil Transmigration Under Flow Conditions

Neutrophils are the most abundant type of white blood cell. They form an essential part of the innate immune system1. During acute inflammation, ...

University of Calgary 

Sialooligosaccharides were generated by direct enzymatic glycosylation on arrays and the resulting surfaces were suitable for the study of carbohydrate-specific cell adhesion.

Chemoenzymatic synthesis of sialooligosaccharides on arrays for studies of cell surface adhesion.

During an inflammatory response, endothelial cells undergo morphological changes to allow for the passage of neutrophils from the blood vessel to the site of injury or infection. Although endothelial cell junctions and the cytoskeleton undergo reorganization during inflammation, little is known about another class of cellular structures, the focal adhesions. In this study, we examined several focal adhesion proteins during an inflammatory response. We found that there was selective loss of paxillin and focal adhesion kinase (FAK) from focal adhesions in proximity to transmigrating neutrophils; in contrast the levels of the focal adhesion proteins β1-integrin and vinculin were unaffected. Paxillin was lost from focal adhesions during neutrophil transmigration both under static and flow conditions. Down-regulating endothelial paxillin with siRNA blocked neutrophil transmigration while having no effect on rolling or adhesion. As paxillin dynamics are regulated partly by FAK, the role of FAK in neutrophil transmigration was examined using two complementary methods. siRNA was used to down-regulate total FAK protein while dominant-negative, kinase-deficient FAK was expressed to block FAK signaling. Disruption of the FAK protein or FAK signaling decreased neutrophil transmigration. Collectively, these findings reveal a novel role for endothelial focal adhesion proteins paxillin and FAK in regulating neutrophil transmigration.

Article	Year
Annals of biomedical engineering\| PubMed ID: 12051621	2002
Inorganic chemistry\| PubMed ID: 14632486	2003
Inorganic chemistry\| PubMed ID: 15762717	2005
Biochemical and biophysical research communications\| PubMed ID: 16298340	2006
The FEBS journal\| PubMed ID: 16704412	2006
The Journal of biological chemistry\| PubMed ID: 16895906	2006
Journal of the Indian Medical Association\| PubMed ID: 17388006	2006
The Kaohsiung journal of medical sciences\| PubMed ID: 17525004	2007
Biochemistry\| PubMed ID: 17536835	2007
Journal of bacteriology\| PubMed ID: 18390660	2008
Anesthesia and analgesia\| PubMed ID: 18713916	2008
Evidence report/technology assessment\| PubMed ID: 20629477	2009
Indian journal of experimental biology\| PubMed ID: 21299043	2010
Chemoenzymatic synthesis of sialooligosaccharides on arrays for studies of cell surface adhesion. Chemical communications (Cambridge, England)\| PubMed ID: 21468399	2011
British journal of hospital medicine (London, England : 2005)\| PubMed ID: 21475096	2011
British journal of hospital medicine (London, England : 2005)\| PubMed ID: 21647051	2011
Endothelial paxillin and focal adhesion kinase (FAK) play a critical role in neutrophil transmigration. European journal of immunology\| PubMed ID: 22095445	2012
Journal of the American Psychiatric Nurses Association\| PubMed ID: 22142975	2011
Gene\| PubMed ID: 22521868	2012

Ritu Sharma

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