A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding
We have adapted an in vitro model of the human blood-brain barrier, the immortalized human cerebral microvascular endothelial cells (hCMEC/D3), to quantitatively measure protein transcytosis. After validating the receptor-mediated transport using transferrin, the system was used to measure transcyto...
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| Veröffentlicht in: | PloS one 2014-04, Vol.9 (4), p.e96340-e96340 |
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| creator | Sade, Hadassah Baumgartner, Claudia Hugenmatter, Adrian Moessner, Ekkehard Freskgård, Per-Ola Niewoehner, Jens |
| description | We have adapted an in vitro model of the human blood-brain barrier, the immortalized human cerebral microvascular endothelial cells (hCMEC/D3), to quantitatively measure protein transcytosis. After validating the receptor-mediated transport using transferrin, the system was used to measure transcytosis rates of antibodies directed against potential brain shuttle receptors. While an antibody to the insulin-like growth factor 1 receptor (IGF1R) was exclusively recycled to the apical compartment, the fate of antibodies to the transferrin receptor (TfR) was determined by their relative affinities at extracellular and endosomal pH. An antibody with reduced affinity at pH5.5 showed significant transcytosis, while pH-independent antibodies of comparable affinities at pH 7.4 remained associated with intracellular vesicular compartments and were finally targeted for degradation. |
| doi_str_mv | 10.1371/journal.pone.0096340 |
| format | Article |
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After validating the receptor-mediated transport using transferrin, the system was used to measure transcytosis rates of antibodies directed against potential brain shuttle receptors. While an antibody to the insulin-like growth factor 1 receptor (IGF1R) was exclusively recycled to the apical compartment, the fate of antibodies to the transferrin receptor (TfR) was determined by their relative affinities at extracellular and endosomal pH. An antibody with reduced affinity at pH5.5 showed significant transcytosis, while pH-independent antibodies of comparable affinities at pH 7.4 remained associated with intracellular vesicular compartments and were finally targeted for degradation.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0096340</identifier><identifier>PMID: 24788759</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Affinity ; Analysis ; Antibodies ; Antibodies - immunology ; Antibodies - metabolism ; Antigens, CD - immunology ; Biology and Life Sciences ; Blood-brain barrier ; Blood-Brain Barrier - metabolism ; Brain research ; Cell Line ; Chemical properties ; Compartments ; Crystal structure ; Endocytosis ; Endothelial cells ; Health aspects ; Humans ; Hydrogen ions ; Hydrogen-Ion Concentration ; Immunoglobulins ; Insulin ; Insulin-like growth factors ; Ligands ; Medicine and Health Sciences ; Microvasculature ; pH effects ; Proteins ; Receptor, IGF Type 1 - immunology ; Receptors ; Receptors, Transferrin - immunology ; Rodents ; Transcytosis ; Transferrin ; Viral antibodies</subject><ispartof>PloS one, 2014-04, Vol.9 (4), p.e96340-e96340</ispartof><rights>COPYRIGHT 2014 Public Library of Science</rights><rights>2014 Sade et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2014 Sade et al 2014 Sade et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-2a4007190bd1fb6f95c12f9fa55ccf44fcda80d534e16876e16432cc7775ab373</citedby><cites>FETCH-LOGICAL-c758t-2a4007190bd1fb6f95c12f9fa55ccf44fcda80d534e16876e16432cc7775ab373</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4005765/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4005765/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23847,27903,27904,53769,53771,79346,79347</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24788759$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Schlachetzki, Felix</contributor><creatorcontrib>Sade, Hadassah</creatorcontrib><creatorcontrib>Baumgartner, Claudia</creatorcontrib><creatorcontrib>Hugenmatter, Adrian</creatorcontrib><creatorcontrib>Moessner, Ekkehard</creatorcontrib><creatorcontrib>Freskgård, Per-Ola</creatorcontrib><creatorcontrib>Niewoehner, Jens</creatorcontrib><title>A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>We have adapted an in vitro model of the human blood-brain barrier, the immortalized human cerebral microvascular endothelial cells (hCMEC/D3), to quantitatively measure protein transcytosis. After validating the receptor-mediated transport using transferrin, the system was used to measure transcytosis rates of antibodies directed against potential brain shuttle receptors. While an antibody to the insulin-like growth factor 1 receptor (IGF1R) was exclusively recycled to the apical compartment, the fate of antibodies to the transferrin receptor (TfR) was determined by their relative affinities at extracellular and endosomal pH. An antibody with reduced affinity at pH5.5 showed significant transcytosis, while pH-independent antibodies of comparable affinities at pH 7.4 remained associated with intracellular vesicular compartments and were finally targeted for degradation.</description><subject>Affinity</subject><subject>Analysis</subject><subject>Antibodies</subject><subject>Antibodies - immunology</subject><subject>Antibodies - metabolism</subject><subject>Antigens, CD - immunology</subject><subject>Biology and Life Sciences</subject><subject>Blood-brain barrier</subject><subject>Blood-Brain Barrier - metabolism</subject><subject>Brain research</subject><subject>Cell Line</subject><subject>Chemical properties</subject><subject>Compartments</subject><subject>Crystal structure</subject><subject>Endocytosis</subject><subject>Endothelial cells</subject><subject>Health aspects</subject><subject>Humans</subject><subject>Hydrogen ions</subject><subject>Hydrogen-Ion Concentration</subject><subject>Immunoglobulins</subject><subject>Insulin</subject><subject>Insulin-like growth factors</subject><subject>Ligands</subject><subject>Medicine and Health Sciences</subject><subject>Microvasculature</subject><subject>pH effects</subject><subject>Proteins</subject><subject>Receptor, IGF Type 1 - immunology</subject><subject>Receptors</subject><subject>Receptors, Transferrin - immunology</subject><subject>Rodents</subject><subject>Transcytosis</subject><subject>Transferrin</subject><subject>Viral antibodies</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk11r2zAUhs3YWLtu_2BshsHYLpxJlmTJN4NQtjVQKOzrVsj6SFQcyZPk0vz7KY1b4tGLYZAl-Xlf-RydUxSvIVhAROGnaz8GJ_rF4J1eANA2CIMnxSlsUV01NUBPj-YnxYsYrwEgiDXN8-KkxpQxStrT4nZZbsatcGXXe6-qLgib5yIEq0OZgnBR7pKPNpYiRrErg77Ros8rl2zn1W7OWGf6UTupVdntyuGiUnrQTmmXslDqIflQdtYp69Yvi2cmG-lX0_us-PX1y8_zi-ry6tvqfHlZSUpYqmqBAaCwBZ2CpmtMSySsTWsEIVIajI1UggFFENawYbTJI0a1lJRSIjpE0Vnx9uA79D7yKWmRQ1IDiOuW4UysDoTy4poPwW5F2HEvLL_b8GHNRUhW9pozojoBcdMYUGMlhSC1ZBRphQxjCO29Pk-njd1WK5kjD6Kfmc6_OLvha3_Dc5SENiQbfJgMgv8z6pj41kap-1447ce7_4aoxqxmGX33D_p4dBO1FjmAfEE-nyv3pnyJKCGMIAQytXiEyo_SWytzhRmb92eCjzNBZpK-TWsxxshXP77_P3v1e86-P2I3udbSJvp-TNa7OAfxAZTBxxi0eUgyBHzfIPfZ4PsG4VODZNmb4wt6EN13BPoLqxANBA</recordid><startdate>20140401</startdate><enddate>20140401</enddate><creator>Sade, Hadassah</creator><creator>Baumgartner, Claudia</creator><creator>Hugenmatter, Adrian</creator><creator>Moessner, Ekkehard</creator><creator>Freskgård, Per-Ola</creator><creator>Niewoehner, Jens</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20140401</creationdate><title>A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding</title><author>Sade, Hadassah ; Baumgartner, Claudia ; Hugenmatter, Adrian ; Moessner, Ekkehard ; Freskgård, Per-Ola ; Niewoehner, Jens</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c758t-2a4007190bd1fb6f95c12f9fa55ccf44fcda80d534e16876e16432cc7775ab373</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Affinity</topic><topic>Analysis</topic><topic>Antibodies</topic><topic>Antibodies - 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After validating the receptor-mediated transport using transferrin, the system was used to measure transcytosis rates of antibodies directed against potential brain shuttle receptors. While an antibody to the insulin-like growth factor 1 receptor (IGF1R) was exclusively recycled to the apical compartment, the fate of antibodies to the transferrin receptor (TfR) was determined by their relative affinities at extracellular and endosomal pH. An antibody with reduced affinity at pH5.5 showed significant transcytosis, while pH-independent antibodies of comparable affinities at pH 7.4 remained associated with intracellular vesicular compartments and were finally targeted for degradation.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24788759</pmid><doi>10.1371/journal.pone.0096340</doi><oa>free_for_read</oa></addata></record> |
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| subjects | Affinity Analysis Antibodies Antibodies - immunology Antibodies - metabolism Antigens, CD - immunology Biology and Life Sciences Blood-brain barrier Blood-Brain Barrier - metabolism Brain research Cell Line Chemical properties Compartments Crystal structure Endocytosis Endothelial cells Health aspects Humans Hydrogen ions Hydrogen-Ion Concentration Immunoglobulins Insulin Insulin-like growth factors Ligands Medicine and Health Sciences Microvasculature pH effects Proteins Receptor, IGF Type 1 - immunology Receptors Receptors, Transferrin - immunology Rodents Transcytosis Transferrin Viral antibodies |
| title | A human blood-brain barrier transcytosis assay reveals antibody transcytosis influenced by pH-dependent receptor binding |
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