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Monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay
We describe a quantitative assay of transendothelial migration (TEM) that allows us to selectively study the interaction of monocytes with confluent human endothelial cell (HEC) monolayers. The HEC are grown on hydrated collagen gels; the monocytes need not be purified. 100% of monocytes transmigrat...
Formato: | Texto |
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Lenguaje: | English |
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The Rockefeller University Press
1992
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2119361/ https://www.ncbi.nlm.nih.gov/pubmed/1512545 |
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collection | PubMed |
description | We describe a quantitative assay of transendothelial migration (TEM) that allows us to selectively study the interaction of monocytes with confluent human endothelial cell (HEC) monolayers. The HEC are grown on hydrated collagen gels; the monocytes need not be purified. 100% of monocytes transmigrated the monolayer within 1 h at 37 degrees C and accumulated in the subendothelial collagen; TEM of lymphocytes was not detected within this time. Migration of neutrophils from the same donor was much slower and incomplete, with only 14% of PMN transmigrating in 2 h. This rapid TEM occurs in the absence of exogenous chemoattractants, and HEC in this system do not express cytokine- inducible leukocyte adhesion molecules. A slight modification of the TEM assay allowed us to separate binding to the apical HEC surface from TEM. We found that tight apical surface binding was the rate-limiting step for TEM. Two-thirds of this binding and TEM could be blocked by a monoclonal antibody against the leukocyte beta 2 integrin chain CD18. This assay will allow us to dissect the mechanisms of both the binding and transmigration stages of diapedesis. |
format | Text |
id | pubmed-2119361 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1992 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21193612008-04-16 Monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay J Exp Med Articles We describe a quantitative assay of transendothelial migration (TEM) that allows us to selectively study the interaction of monocytes with confluent human endothelial cell (HEC) monolayers. The HEC are grown on hydrated collagen gels; the monocytes need not be purified. 100% of monocytes transmigrated the monolayer within 1 h at 37 degrees C and accumulated in the subendothelial collagen; TEM of lymphocytes was not detected within this time. Migration of neutrophils from the same donor was much slower and incomplete, with only 14% of PMN transmigrating in 2 h. This rapid TEM occurs in the absence of exogenous chemoattractants, and HEC in this system do not express cytokine- inducible leukocyte adhesion molecules. A slight modification of the TEM assay allowed us to separate binding to the apical HEC surface from TEM. We found that tight apical surface binding was the rate-limiting step for TEM. Two-thirds of this binding and TEM could be blocked by a monoclonal antibody against the leukocyte beta 2 integrin chain CD18. This assay will allow us to dissect the mechanisms of both the binding and transmigration stages of diapedesis. The Rockefeller University Press 1992-09-01 /pmc/articles/PMC2119361/ /pubmed/1512545 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Articles Monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay |
title | Monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay |
title_full | Monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay |
title_fullStr | Monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay |
title_full_unstemmed | Monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay |
title_short | Monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay |
title_sort | monocyte-selective transendothelial migration: dissection of the binding and transmigration phases by an in vitro assay |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2119361/ https://www.ncbi.nlm.nih.gov/pubmed/1512545 |