A blocking antibody to the hyaluronan receptor for endocytosis (HARE) inhibits hyaluronan clearance by perfused liver.
Hyaluronan (HA) and chondroitin sulfate clearance from lymph and blood is mediated by the hyaluronan receptor for endocytosis (HARE). The purification and molecular cloning (Zhou, B., Weigel, J. A., Saxena, A., and Weigel, P. H. (2002) Mol. Biol. Cell 13, 2853-2868) of this cell surface receptor were finally achieved after we developed monoclonal antibodies (mAbs) against HARE. There are actually two independent isoreceptors for HA, which in rat are designated the 175-kDa HARE and 300-kDa HARE. Only one mAb (number 174) effectively and completely blocked the specific uptake of 125I-HA at 37 degrees C by rat liver sinusoidal endothelial cells. 125I-HA binding to both the 175-kDa and 300-kDa HARE proteins in a ligand blot assay was almost completely inhibited by <1 microg/ml mAb-174, whereas mouse IgG had little or no effect. MAb-174 also performed very well in Western analysis, indirect fluorescence microscopy, and a variety of immuno-procedures. Immunohistochemistry using mAb-174 localized HARE to the sinusoidal cells of rat liver, spleen, and lymph node. Western analysis using mAb-174 revealed that the sizes of both HARE glycoproteins were the same in these three tissues. 125I-HA was taken up and degraded by excised rat livers that were continuously perfused ex vivo with a recirculating medium. This HA clearance and metabolism by liver, which is a physiological function of HARE, was very effectively blocked by mAb-174 but not by mouse IgG. The results indicate that mAb-174 will be a useful tool to study the functions of HARE and the physiological significance of HA clearance.
Weigel
Janet A
JA
Department of Biochemistry and Molecular Biology and the Oklahoma Center for Medical Glycobiology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73190, USA.
Raymond
Robert C
RC
McGary
Carl
C
Singh
Anil
A
Weigel
Paul H
PH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism; D015153:Blotting, Western; D002478:Cells, Cultured; D004306:Dose-Response Relationship, Immunologic; D004727:Endothelium / Q000166:cytology; D018960:Hyaluronan Receptors / Q000737:chemistry* / Q000378:metabolism*; D006820:Hyaluronic Acid / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007074:Immunoglobulin G / Q000378:metabolism; D007150:Immunohistochemistry; D008024:Ligands; D008099:Liver / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D008198:Lymph Nodes / Q000166:cytology; D008297:Male; D051379:Mice; D018613:Microscopy, Confocal; D010477:Perfusion; D051381:Rats; D017207:Rats, Sprague-Dawley; D013154:Spleen / Q000166:cytology; D013997:Time Factors
D000911:Antibodies, Monoclonal; D018960:Hyaluronan Receptors; D007074:Immunoglobulin G; D008024:Ligands; C472814:hyaluronan receptor 175-kDa, rat; D006820:Hyaluronic Acid