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Nephrology Dialysis Transplantation, Vol 13, Issue 2 269-276, Copyright © 1998 by Oxford University Press

ORIGINAL ARTICLES

Characterization of CD44-mediated hyaluronan binding by renal tubular epithelial cells

B Oertli, X Fan and R Wuthrich
Physiological Institute, University of Zurich-Irchel, Zurich, Switzerland; Division of Nephrology, University Hospital, Ramistrasse 100, CH-8091 Zurich, Switzerland; Corresponding author address

Background. CD44 is the main receptor for the extracellular polysaccharide hyaluronan (HA). We have recently shown that CD44 is strongly induced on renal tubular epithelial cells (TEC) in autoimmune renal injury and that HA accumulates in the renal interstitium (Kidney Int 1996; 50: 156-163 and Nephrol Dial Transplant 1997; 12: 1344-1353). The functional significance of enhanced tubular CD44 expression and its interaction with HA are not known. The purpose of the present study was to characterize renal tubular CD44 expression and CD44-mediated HA binding in vitro and to investigate the growth modulating effects in response to HA binding by TEC. Methods. RT-PCR analysis, flow cytometry, confocal microscopy and Western blotting were used to examine cell surface and soluble CD44 expression by cultured TEC, using SV40-transformed mouse cortical tubular (MCT) cells. HA binding characteristics were examined by flow cytometry and effects of HA on TEC cell growth by [3H]thymidine incorporation. Results. By RT-PCR analysis MCT cells expressed predominantly the standard form of CD44 mRNA, whereas the expression of variant forms was very weak. Confocal microscopy showed that CD44 was expressed basolaterally and apically on MCT cells with strong staining on microvilli. Shedding of CD44 from MCT cells could be induced with crosslinking of anti-CD44 mAbs or with PMA stimulation. MCT cells constitutively bound HA and this binding could be modulated with anti-CD44 mAbs. Soluble and plate-bound HA markedly inhibited MCT cell growth. Conclusions. CD44 is a regulated HA receptor on MCT cells which can be shed into the cellular environment. Upon binding of HA, CD44 functions as a growth inhibitory cell surface protein in MCT cells. We speculate that the interaction of CD44 with HA may have important regulatory effects on cell proliferation in tubulointerstitial renal diseases. Keywords: CD44; cell growth; hyaluronan; tubular epithelial cells
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