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Published on: June 29, 2016
DC-SIGN binds ICAM-3 isolated from peripheral human leukocytes through Lewis x residues
Valentina Bogoevska1, Peter Nollau, Lothar Lucka
1Institut für Klinische Chemie, Universitätsklinikum Hamburg-Eppendorf, Martinistrasse 52, D-20251 Hamburg, Germany.
Insights
Intercellular adhesion molecule-3 (ICAM-3) binds to dendritic cell-specific ICAM-3 grabbing nonintegrin (DC-SIGN) via Lewis x residues, primarily synthesized by fucosyltransferase IX in myeloid cells. This interaction facilitates granulocyte engagement with dendritic cells.
Area of Science:
- Immunology
- Glycobiology
- Cell Biology
Background:
- Intercellular adhesion molecule-3 (ICAM-3) facilitates T cell and antigen-presenting cell interactions via alpha(L)beta(2) integrin.
- Dendritic cell-specific ICAM-3 grabbing nonintegrin (DC-SIGN) is a C-type lectin on macrophages and dendritic cells, potentially binding ICAM-3.
- The specific glycan structure of ICAM-3 involved in DC-SIGN binding remains unidentified.
Purpose of the Study:
- To identify the glycan structure of ICAM-3 responsible for binding to DC-SIGN.
- To determine which fucosyltransferases synthesize the relevant glycan structures on ICAM-3.
- To elucidate the role of ICAM-3 in granulocyte interactions with dendritic cells.
Main Methods:
- Binding assays using recombinant DC-SIGN and native ICAM-3 from human peripheral leukocytes.
- Recognition by monoclonal Lewis x antibodies.
- Mass spectrometry (MALDI-TOF) to confirm Lewis x residues.
- Cotransfection studies with different fucosyltransferase (FUT) genes (FUT IX, IV, III, VII).
Main Results:
- Native ICAM-3 binds recombinant DC-SIGN and immature dendritic cells.
- Lewis x residues are present on ICAM-3, confirmed by mass spectrometry.
- Only ICAM-3 from granulocytes bound DC-SIGN.
- Fucosyltransferase IX (FUT IX) and, to a lesser extent, FUT IV, mediate Lewis x synthesis on ICAM-3.
- FUT IX is the primary enzyme for Lewis x synthesis on ICAM-3 in myeloid cells.
Conclusions:
- Lewis x residues on ICAM-3 are the ligands for DC-SIGN.
- FUT IX is the main enzyme responsible for generating these Lewis x epitopes on ICAM-3 in myeloid cells.
- ICAM-3 binding to DC-SIGN, mediated by Lewis x, likely contributes to granulocyte-dendritic cell interactions.
Abstract:
Intercellular adhesion molecule-3 (ICAM-3) binds to the alpha(L)beta(2) integrin and mediates the contact between T cells and antigen-presenting cells. It has been suggested that dendritic cell-specific ICAM-3 grabbing nonintegrin (DC-SIGN), a C-type lectin of macrophages and DCs, is an additional ligand of ICAM-3. So far, the glycan structure mediating the interaction of native ICAM-3 with DC-SIGN is undefined. Here, we demonstrate that native ICAM-3 from human peripheral leukocytes binds recombinant DC-SIGN, is recognized by monoclonal Lewis x antibodies, and specifically interacts with DC-SIGN on immature DCs. The presence of Lewis x residues on ICAM-3 was confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy. Investigations on different peripheral blood cell populations revealed that only ICAM-3 from granulocytes bound DC-SIGN. Cotransfection studies demonstrated that fucosyltransferase (FUT) IX and, to a significantly lesser extent, FUT IV, but not FUTs III and VII, mediate the synthesis of Lewis x residues on ICAM-3. These findings indicate that FUT IX is the main FUT mediating the synthesis of Lewis x residues of ICAM-3 in cells of the myeloid lineage, and that these residues bind DC-SIGN. The results suggest that ICAM-3 assists in the interaction of granulocytes with DC-SIGN of DCs.
