Interaction of RANTES with syndecan-1 and syndecan-4 expressed by human primary macrophages

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Abstract

Interaction of RANTES with its membrane ligands or receptors transduces multiple intracellular signals. Whether RANTES uses proteoglycans (PGs) belonging to the syndecan family to attach to primary cells expressing RANTES G-protein-coupled receptors (GPCRs) was investigated. We demonstrate that RANTES specifically binds to high and low affinity binding sites on human monocyte-derived macrophages (MDM). We show by co-immunoprecipitation experiments that RANTES is associated on these cells with syndecan-1 and syndecan-4, but neither with syndecan-2 nor with betaglycan, in addition to CD44 and its GPCRs, CCR5 and CCR1. Glycosaminidases pre-treatment of the monocyte derived-macrophages strongly decreases the binding of RANTES to syndecan-1 and syndecan-4 and also to CCR5, and abolishes RANTES binding to CD44. This suggests that glycosaminoglycans (GAGs) are involved in RANTES binding to the PGs and that such bindings facilitate the subsequent interaction of RANTES with CCR5, on the MDM, characterized by low membrane expression of CCR5. The role of these interactions in the pathophysiology of RANTES deserves further study.

Keywords

RANTES
CCR5
Syndecan
HIV
Macrophage

Abbreviations

MDM
monocyte-derived macrophages
PBL
peripheral blood lymphocytes
PGs
proteoglycans
SD
syndecan
MIP
macrophage inflammatory protein
MCP-1
monocyte chemoattractant protein-1
RANTES
regulated on activation normal T cell expressed and secreted
GAGs
glycosaminoglycans
SDF
stromal-cell derived factor
GPCRs
G-protein-coupled receptors
BSA
bovine serum albumin
PBS
phosphate-buffered saline
DTT
dithiothreitol
TBS
Tris-buffered saline supplemented
FCS
fetal calf serum
IL-8
interleukin-8
FGF-2
fibroblast growth factor-2
hbEGF
heparin-binding epidermal growth factor

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