Postcapillary venule endothelial cells in kidney express a multispecific chemokine receptor that is structurally and functionally identical to the erythroid isoform, which is the Duffy blood group antigen.
The human erythrocyte chemokine receptor has recently been shown to be identical to the Duffy blood group antigen and is expressed in multiple organs, including kidney. Here we have examined the molecular properties of the renal isoform. Immunoblot analysis of erythrocyte and kidney detergent lysates, with a monoclonal antibody (Fy6) to the Duffy antigen, revealed that the renal isoform had a molecular mass of 43-45 kD, which could be distinguished from that observed in erythroid cells (38-47 kD). Chemical cross-linking of kidney membranes to 125I-melanoma growth stimulatory activity (MGSA) indicated that the renal chemokine receptor had a molecular mass of 38-45 kD. Binding of 125I-labeled MGSA to kidney membranes was competitively inhibited by the addition of unlabeled MGSA, IL-8, regulated on activation, normal T expressed and secrted, and monocyte chemotactic protein-1. Scatchard analysis of MGSA binding showed that the chemokine receptor from renal tissues had a binding affinity of 3.5 nM similar to that observed for the erythroid isoform (5-10 nM). The primary structure of the renal chemokine receptor predicted from the nucleotide sequence of cDNA from renal tissues is identical to that reported for the erythroid isoform. Immunocytochemical staining of kidney with Fy6 localized expression to endothelial cells present in postcapillary venules. These studies implicate the Duffy antigen/chemokine receptor in the complex interactions between postcapillary endothelial cells and granulocytes, which are modulated by pro-inflammatory chemokines.
Hadley
T J
TJ
Henry Vogt Cancer Research Institute, James Graham Brown Cancer Center, Department of Medicine, University of Louisville School of Medicine, Kentucky 40292.
Lu
Z H
ZH
Wasniowska
K
K
Martin
A W
AW
Peiper
S C
SC
Hesselgesser
J
J
Horuk
R
R
D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000378:metabolism; D004375:Duffy Blood-Group System / Q000302:isolation & purification / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D015870:Gene Expression; D006133:Growth Substances / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins; D007700:Kinetics; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D018121:Receptors, Cytokine / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D012079:Renal Circulation; D014699:Venules
D000911:Antibodies, Monoclonal; C475829:CXCL1 protein, human; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors; D004375:Duffy Blood-Group System; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D018121:Receptors, Cytokine; C078862:melanoma growth stimulating activity receptor