| 2E8 binds to the high affinity I-domain in a metal ion-dependent manner: a second generation monoclonal antibody selectively targeting activated LFA-1. | |
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MedLine Citation:
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PMID: 20724473 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The activation of leukocyte function-associated antigen-1 (LFA-1) plays a critical role in regulating immune responses. The metal ion-dependent adhesion site on the I-domain of LFA-1 α(L) subunit is the key recognition site for ligand binding. Upon activation, conformation changes in the I-domain can lead LFA-1 from the low affinity state to the high affinity (HA) state. Using the purified HA I-domain locked by disulfide bonds for immunization, we developed an mAb, 2E8, that specifically binds to cells expressing the HA LFA-1. The surface plasmon resonance analysis has shown that 2E8 only binds to the HA I-domain and that the dissociation constant (K(D)) for HA I-domain is 197 nm. The binding of 2E8 to the HA I-domain is metal ion-dependent, and the affinity decreased as Mn(2+) was replaced sequentially by Mg(2+) and Ca(2+). Surface plasmon resonance analysis demonstrates that 2E8 inhibits the interaction of HA I-domain and ICAM-1. Furthermore, we found that 2E8 can detect activated LFA-1 on both JY and Jurkat cells using flow cytometry and parallel plate adhesion assay. In addition, 2E8 inhibits JY cell adhesion to human umbilical vein endothelial cells and homotypic aggregation. 2E8 treatment reduces the proliferation of both human CD4(+) and CD8(+) T cells upon OKT3 stimulation without the impairment of their cytolytic function. Taken together, these data demonstrate that 2E8 is specific for the high affinity form of LFA-1 and that 2E8 inhibits LFA-1/ICAM-1 interactions. As a novel activation-specific monoclonal antibody, 2E8 is a potentially useful reagent for blocking high affinity LFA-1 and modulating T cell activation in research and therapeutics. |
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Authors:
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Roberto Carreño; Wells S Brown; Dan Li; Jessica A Hernandez; Yang Wang; Tae Kon Kim; John W Craft; Krishna V Komanduri; Laszlo G Radvanyi; Patrick Hwu; Jeffrey J Molldrem; Glen B Legge; Bradley W McIntyre; Qing Ma |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-08-19 |
Journal Detail:
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Title: The Journal of biological chemistry Volume: 285 ISSN: 1083-351X ISO Abbreviation: J. Biol. Chem. Publication Date: 2010 Oct |
Date Detail:
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Created Date: 2010-10-18 Completed Date: 2010-11-24 Revised Date: 2011-10-24 |
Medline Journal Info:
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Nlm Unique ID: 2985121R Medline TA: J Biol Chem Country: United States |
Other Details:
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Languages: eng Pagination: 32860-8 Citation Subset: IM |
Affiliation:
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Section of Transplantation Immunology, Department of Stem Cell Transplantation and Cellular Therapy, University of Texas MD Anderson Cancer Center,Houston, Texas 77030, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Antibodies, Monoclonal / immunology, pharmacology* CD4-Positive T-Lymphocytes / cytology, immunology*, metabolism CD8-Positive T-Lymphocytes / cytology, immunology*, metabolism Cations, Divalent / immunology, metabolism Cell Adhesion / drug effects, immunology Disulfides / immunology, metabolism Endothelial Cells / cytology, immunology, metabolism Flow Cytometry Gene Expression Regulation / drug effects, immunology Humans Immunity, Cellular / drug effects, immunology Intercellular Adhesion Molecule-1 / immunology*, metabolism Jurkat Cells K562 Cells Lymphocyte Activation / drug effects, immunology Lymphocyte Function-Associated Antigen-1 / immunology*, metabolism Metals / immunology*, metabolism Mice Mice, Inbred BALB C Muromonab-CD3 / immunology, metabolism Protein Structure, Tertiary Protein Subunits / immunology, metabolism Surface Plasmon Resonance / methods Umbilical Veins / cytology, immunology |
| Chemical | |
Reg. No./Substance:
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0/Antibodies, Monoclonal; 0/Cations, Divalent; 0/Disulfides; 0/Lymphocyte Function-Associated Antigen-1; 0/Metals; 0/Muromonab-CD3; 0/Protein Subunits; 126547-89-5/Intercellular Adhesion Molecule-1 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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