CCL22-specific Antibodies Reveal That Engagement of Two Distinct Binding Domains on CCL22 Is Required for CCR4-mediated Function
CCL22 inactivation in vivo occurs by cleavage at the N-terminus; however, it is unclear whether this encompasses the entire site of CCR4 interaction. CCL17 also binds CCR4 and its function requires binding via two discrete binding sites. Using monoclonal antibodies (MAbs), we report that there are t...
Saved in:
Published in | Monoclonal antibodies in immunodiagnosis and immunotherapy Vol. 34; no. 6; pp. 373 - 380 |
---|---|
Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Mary Ann Liebert, Inc
01.12.2015
|
Subjects | |
Online Access | Get full text |
Cover
Loading…
Abstract | CCL22 inactivation
in vivo
occurs by cleavage at the N-terminus; however, it is unclear whether this encompasses the entire site of CCR4 interaction. CCL17 also binds CCR4 and its function requires binding via two discrete binding sites. Using monoclonal antibodies (MAbs), we report that there are two separate sites on CCL22 that are required for CCR4-mediated function. The CCL22-specific antibodies bind with affinities of 632 ± 297 pM (MC2B7) and 308 ± 43 pM (MAB4391) and neither exhibited detectable binding to CCL17. Both antibodies are comparable in their ability to inhibit CCL22-mediated calcium mobilization; however, competition binding studies demonstrate that MC2B7 and MAB4391 bind to distinct epitopes on CCL22. Both antibodies inhibit function through CCR4, which is demonstrated by loss of β-arrestin recruitment in a reporter cell line. In both assays, blocking either site independently abolished CCL22 function, suggesting that concurrent engagement of both sites with CCR4 is necessary for function. This is the first demonstration that CCL22 has two distinct binding sites that are required for CCR4 function. These antibodies are valuable tools for better understanding the interaction and function of CCL22 and CCR4 and will potentially help further understanding of the differential outcomes of CCL17 and CCL22 interaction with CCR4. |
---|---|
AbstractList | CCL22 inactivation in vivo occurs by cleavage at the N-terminus; however, it is unclear whether this encompasses the entire site of CCR4 interaction. CCL17 also binds CCR4 and its function requires binding via two discrete binding sites. Using monoclonal antibodies (MAbs), we report that there are two separate sites on CCL22 that are required for CCR4-mediated function. The CCL22-specific antibodies bind with affinities of 632 ± 297 pM (MC2B7) and 308 ± 43 pM (MAB4391) and neither exhibited detectable binding to CCL17. Both antibodies are comparable in their ability to inhibit CCL22-mediated calcium mobilization; however, competition binding studies demonstrate that MC2B7 and MAB4391 bind to distinct epitopes on CCL22. Both antibodies inhibit function through CCR4, which is demonstrated by loss of β-arrestin recruitment in a reporter cell line. In both assays, blocking either site independently abolished CCL22 function, suggesting that concurrent engagement of both sites with CCR4 is necessary for function. This is the first demonstration that CCL22 has two distinct binding sites that are required for CCR4 function. These antibodies are valuable tools for better understanding the interaction and function of CCL22 and CCR4 and will potentially help further understanding of the differential outcomes of CCL17 and CCL22 interaction with CCR4. CCL22 inactivation in vivo occurs by cleavage at the N-terminus; however, it is unclear whether this encompasses the entire site of CCR4 interaction. CCL17 also binds CCR4 and its function requires binding via two discrete binding sites. Using monoclonal antibodies (MAbs), we report that there are two separate sites on CCL22 that are required for CCR4-mediated function. The CCL22-specific antibodies bind with affinities of 632 ± 297 pM (MC2B7) and 308 ± 43 pM (MAB4391) and neither exhibited detectable binding to CCL17. Both antibodies are comparable in their ability to inhibit CCL22-mediated calcium mobilization; however, competition binding studies demonstrate that MC2B7 and MAB4391 bind to distinct epitopes on CCL22. Both antibodies inhibit function through CCR4, which is demonstrated by loss of β-arrestin recruitment in a reporter cell line. In both assays, blocking either site independently abolished CCL22 function, suggesting that concurrent engagement of both sites with CCR4 is necessary for function. This is the first demonstration that CCL22 has two distinct binding sites that are required for CCR4 function. These antibodies are valuable tools for better understanding the interaction and function of CCL22 and CCR4 and will potentially help further understanding of the differential outcomes of CCL17 and CCL22 interaction with CCR4. |
Author | Lacy, Eilyn R. Luongo, Jennifer Santulli-Marotto, Sandra Ryan, Mary Wheeler, John Boakye, Ken Wu, Sheng-Jiun |
Author_xml | – sequence: 1 givenname: Sandra surname: Santulli-Marotto fullname: Santulli-Marotto, Sandra – sequence: 2 givenname: John surname: Wheeler fullname: Wheeler, John – sequence: 3 givenname: Eilyn R. surname: Lacy fullname: Lacy, Eilyn R. – sequence: 4 givenname: Ken surname: Boakye fullname: Boakye, Ken – sequence: 5 givenname: Jennifer surname: Luongo fullname: Luongo, Jennifer – sequence: 6 givenname: Sheng-Jiun surname: Wu fullname: Wu, Sheng-Jiun – sequence: 7 givenname: Mary surname: Ryan fullname: Ryan, Mary |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/26683175$$D View this record in MEDLINE/PubMed |
BookMark | eNpNkM1OwzAQhC1UREvpkSvyC6T4J3HiY0lbqFQJqSrnyI7XxaixS5KCuPHoJBQQe9nVaHY0-i7RwAcPCF1TMqUkk7eV0lNGaDIlhMszNGJUpJGMuRj8u4do0jQvpJuUMsqTCzRkQmScpskIfeb5mrGoOUDprCvxzLdOB-OgwRt4A7XH22fV4oXfqR1U4FscLN6-Bzx3Tet82eI7543zOzwPlXK-wcHj70y86iNej64Gg22oO3UTRxUYp9pOWR67Zxf8FTq3at_A5GeP0dNysc0fovXj_SqfraOSs1hGsSUWaKpTyaiNS2tSqynniYKEi0yApIxnpWRMxDGxQmfaSJlwbUycUW2Aj9HNKfdw1F2J4lC7StUfxS-KzsBPhl5W3u8daKjbPyMlRc-86JgXPfOiZ86_AFWjcs8 |
CitedBy_id | crossref_primary_10_1016_j_drudis_2016_11_023 crossref_primary_10_1038_s41417_023_00678_z crossref_primary_10_4049_jimmunol_2000315 |
ContentType | Journal Article |
Copyright | Sandra Santulli-Marotto et al. 2015; Published by Mary Ann Liebert, Inc. |
Copyright_xml | – notice: Sandra Santulli-Marotto et al. 2015; Published by Mary Ann Liebert, Inc. |
DBID | 1-M CGR CUY CVF ECM EIF NPM |
DOI | 10.1089/mab.2015.0039 |
DatabaseName | Mary Ann Liebert Online - Open Access Medline MEDLINE MEDLINE (Ovid) MEDLINE MEDLINE PubMed |
DatabaseTitle | MEDLINE Medline Complete MEDLINE with Full Text PubMed MEDLINE (Ovid) |
DatabaseTitleList | MEDLINE |
Database_xml | – sequence: 1 dbid: NPM name: PubMed url: https://proxy.k.utb.cz/login?url=http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed sourceTypes: Index Database – sequence: 2 dbid: EIF name: MEDLINE url: https://proxy.k.utb.cz/login?url=https://www.webofscience.com/wos/medline/basic-search sourceTypes: Index Database – sequence: 3 dbid: 1-M name: Mary Ann Liebert Online - Open Access url: http://liebertopenaccess.com/OAJournals sourceTypes: Publisher |
DeliveryMethod | fulltext_linktorsrc |
Discipline | Biology |
EISSN | 2167-9436 |
EndPage | 380 |
ExternalDocumentID | 26683175 10_1089_mab_2015_0039 |
Genre | Journal Article |
GroupedDBID | 0R~ 1-M ABBKN ACGFS ADBBV ALMA_UNASSIGNED_HOLDINGS BNQNF EBS EJD IM4 O9- RML UE5 CGR CUY CVF ECM EIF NPM |
ID | FETCH-LOGICAL-c3249-4f0fe17b7921f4cfd7fb1335ae53686e91238c9226440f6b8bd9953bdd481bde3 |
IEDL.DBID | 1-M |
ISSN | 2167-9436 |
IngestDate | Sat Sep 28 07:58:13 EDT 2024 Fri Sep 27 01:18:48 EDT 2024 |
IsDoiOpenAccess | true |
IsOpenAccess | true |
IsPeerReviewed | true |
IsScholarly | true |
Issue | 6 |
Language | English |
License | This Open Access article is distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. |
LinkModel | DirectLink |
MergedId | FETCHMERGED-LOGICAL-c3249-4f0fe17b7921f4cfd7fb1335ae53686e91238c9226440f6b8bd9953bdd481bde3 |
OpenAccessLink | https://www.liebertpub.com/doi/abs/10.1089/mab.2015.0039 |
PMID | 26683175 |
PageCount | 8 |
ParticipantIDs | pubmed_primary_26683175 maryannliebert_primary_10_1089_mab_2015_0039 |
PublicationCentury | 2000 |
PublicationDate | 20151201 2015-Dec |
PublicationDateYYYYMMDD | 2015-12-01 |
PublicationDate_xml | – month: 12 year: 2015 text: 20151201 day: 01 |
PublicationDecade | 2010 |
PublicationPlace | United States |
PublicationPlace_xml | – name: United States |
PublicationTitle | Monoclonal antibodies in immunodiagnosis and immunotherapy |
PublicationTitleAlternate | Monoclon Antib Immunodiagn Immunother |
PublicationYear | 2015 |
Publisher | Mary Ann Liebert, Inc |
Publisher_xml | – name: Mary Ann Liebert, Inc |
SSID | ssj0000712135 |
Score | 2.0748656 |
Snippet | CCL22 inactivation
in vivo
occurs by cleavage at the N-terminus; however, it is unclear whether this encompasses the entire site of CCR4 interaction. CCL17... CCL22 inactivation in vivo occurs by cleavage at the N-terminus; however, it is unclear whether this encompasses the entire site of CCR4 interaction. CCL17... |
SourceID | pubmed maryannliebert |
SourceType | Index Database Publisher |
StartPage | 373 |
SubjectTerms | Animals Antibodies, Monoclonal - biosynthesis Antibodies, Monoclonal - chemistry Arrestins - genetics Arrestins - immunology beta-Arrestins Binding Sites Binding, Competitive Cell Line Chemokine CCL17 - genetics Chemokine CCL17 - immunology Chemokine CCL22 - genetics Chemokine CCL22 - immunology Dendritic Cells - cytology Dendritic Cells - immunology Epitopes - chemistry Epitopes - genetics Epitopes - immunology Gene Expression Regulation Humans Immunity, Innate Mice Original Articles Protein Binding Protein Structure, Tertiary Receptors, CCR4 - genetics Receptors, CCR4 - immunology Signal Transduction T-Lymphocytes - cytology T-Lymphocytes - immunology |
Title | CCL22-specific Antibodies Reveal That Engagement of Two Distinct Binding Domains on CCL22 Is Required for CCR4-mediated Function |
URI | https://www.liebertpub.com/doi/abs/10.1089/mab.2015.0039 https://www.ncbi.nlm.nih.gov/pubmed/26683175 |
Volume | 34 |
hasFullText | 1 |
inHoldings | 1 |
isFullTextHit | |
isPrint | |
link | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwlV1LS8NAEF5qRVBQfFStL_bg0WCTbB57rH1QxRYpLfQWsi_toRuxUfHmT3cmiQXFi5cQQvg2zO5mvt39ZoaQSx6plhQweGNluMMYbzmp0tppGe27woRKF7UIhqNwMGV3s2BWI_F3LEypa9YoKobvKX7VOLdTsawEcfx6kQqUZOFeiM_XyDqQ6gBrF7jOcLW7Ao7TK6trepjYmzM8o2z8ibBFtjFILLW2avcXwSwcTX-X7FQMkbbLLt0jNW33yUZZM_LjgHx2OvewssMISVT50LbN5yJDLSAd6zegfXTylOa0Zx8rWQvNDJ28Z7SLs9nKnN7Mi0gW2s0W6dwuaWZpgUlvEQKVwVpR4LLwdMycIrQEaCntgwfEXmyQab836QycqoyCI4EtQQcYMLwbiYh7rmHSqMgIWJkGqQ78MA41B-cVS44RtaxlQhELxXngC6UYcFql_UNSt5nVx4SGmO4MIHzGJQMAIV2jU09AA1EoQ9kkVz9tmTyXOTOS4qw75gmYPkHTY05S3iRHpaVXrwFdiJHRnPwP6JRs4n0pMzkj9fzlVZ8DWcjFRTEu4Dp6GH4BYHe7ZA |
link.rule.ids | 315,786,790,21752,27957,27958,55647,55659 |
linkProvider | Mary Ann Liebert, Inc. |
linkToHtml | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwlV3JTsMwELVYhAAJxFL2xQeORGRxnPgIpVWBtoeqSL1F8QY94CAIIG58OjNJhABx4WpF42i8zJvxmxlCTkSifSVh86baCo8x4Xu5NsbzrYkCabk2VS-CwZD3btn1JJ58a_VV85oNkorhf6qrGs82xqIbRpw4e8glcrIwGBKJWTIfx5yh3xV4g6_wCljOsG6vGWJlb8HwkbL1p4RlsoJZYrlzzcS_EGZlabprZLWBiPS8XtN1MmPcBlmom0a-b5KPdrsPrh2mSCLNh567cioLJAPSkXkF3EfH93lJO-6u4bXQwtLxW0Ev8Tg7VdKLaZXKQi-Lh3zqnmnhaCWTXqEIpAYbTQHMwuiIeVVuCeBS2gUTiMvYIrfdzrjd85o-Cp4CuAQrYEHzQSITEQaWKasTK8E1jXMTRzzlRoD1SpXAlFrmWy5TqYWII6k1A1CrTbRF5lzhzA6hHOudgYiICcVAgFSBNXkoYYKEK652yelPXWaPddGMrHrsTkUGqs9Q9ViUVOyS7VrTX58BXkgR0uz9T9AxWeyNB_2sfzW82SdLOF5zTg7IXPn0Yg4BOZTyqNojny-tvco |
linkToPdf | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwlV1LT8MwDI54CAQSiMeA8cyBIxVrmz5yHBsVj21C0yZxq5oX7LAUQQFx46djt9UEiAvXqHIq24m_JJ9tQk55pFpSgPPGynCHMd5yMqW10zLad4UJlS57EfQH4dWY3dwH999afVW8Zo2kYvifcqvGta2flKkZcfx8mgnkZOFliM_nyWIAoAQd2nX6s-sViJxe1V7Tw8renOEjZeNPCatkDbPEMmvriX8hzDLSJBtkvYaItF3ZdJPMabtFlqqmkR_b5LPT6cHRDlMkkeZD27aYiBzJgHSo3wD30dFjVtBL-1DzWmhu6Og9p11czlYW9GJSprLQbj7NJvaF5paWMuk1ikBqsFYUwCyMDplT5pYALqUJhEA0Y4OMk8tR58qp-yg4EuASWMCA5t1IRNxzDZNGRUbA0TTIdOCHcag5RK9YckypZS0TilgozgNfKMUA1Crt75AFm1u9R2iI9c5AhM-4ZCBASNfozBMwQRTKUDbJ2U9dpk9V0Yy0fOyOeQqqT1H1WJSUN8lupenZZ4AXYoQ0-_8TdEKW77pJ2rse3B6QFRyuKCeHZKF4ftVHABwKcVy6yBdE7r1d |
openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=CCL22-specific+Antibodies+Reveal+That+Engagement+of+Two+Distinct+Binding+Domains+on+CCL22+Is+Required+for+CCR4-mediated+Function&rft.jtitle=Monoclonal+antibodies+in+immunodiagnosis+and+immunotherapy&rft.au=Santulli-Marotto%2C+Sandra&rft.au=Wheeler%2C+John&rft.au=Lacy%2C+Eilyn+R.&rft.au=Boakye%2C+Ken&rft.date=2015-12-01&rft.pub=Mary+Ann+Liebert%2C+Inc&rft.issn=2167-9436&rft.eissn=2167-9436&rft.volume=34&rft.issue=6&rft.spage=373&rft.epage=380&rft_id=info:doi/10.1089%2Fmab.2015.0039&rft.externalDocID=10_1089_mab_2015_0039 |
thumbnail_l | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=2167-9436&client=summon |
thumbnail_m | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=2167-9436&client=summon |
thumbnail_s | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=2167-9436&client=summon |