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Am J Physiol Cell Physiol 276: C450-C458, 1999;
0363-6143/99 $5.00
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Vol. 276, Issue 2, C450-C458, February 1999

Hypoxia-induced expression of complement receptor type 1 (CR1, CD35) in human vascular endothelial cells

Charles D. Collard1, Cuneyt Bukusoglu1, Azin Agah1, Sean P. Colgan1, Wende R. Reenstra2, B. Paul Morgan3, and Gregory L. Stahl1

1 Center for Experimental Therapeutics and Reperfusion Injury, Department of Anesthesia, Brigham and Women's Hospital, Harvard Medical School, Boston 02115; 2 Department of Dermatology, Boston University School of Medicine, Boston, Massachusetts 02118; and 3 Department of Medical Biochemistry, University of Wales College of Medicine, Cardiff, United Kingdom

Reoxygenation of hypoxic human umbilical vein endothelial cells (HUVECs) increases protein expression of the complement regulators CD46 and CD55. As the receptor for C3b is known to be present on injured bovine endothelial cells, we investigated whether hypoxia or inflammatory mediators induce complement receptor type 1 (CR1; CD35) expression on HUVECs. CR1 protein expression increased 3.7 ± 0.6-fold as measured by ELISA on HUVECs following hypoxia (48 h, 1% O2). Colocalization of CD35 and von Willebrand factor by confocal microscopy confirmed that CD35 was predominantly intracellular. Lipopolysaccharide or tumor necrosis factor-alpha also significantly increased HUVEC CR1 protein expression. Western blot analysis of neutrophil or hypoxic HUVEC lysates revealed a 221-kDa CR1 band under nonreducing conditions. RT-PCR of hypoxic HUVEC mRNA revealed a single band that, after sequencing, was identified as CD35. In situ hybridization of hypoxic HUVECs, but not normoxic HUVECs or fibroblasts, demonstrated increased CD35 mRNA. Hypoxic HUVECs bound immune complexes and acted as a cofactor for factor I-mediated cleavage of C3b. Thus hypoxia induces functional HUVEC CR1 expression.

cytokines; tumor necrosis factor; human umbilical vein endothelial cells; C3b; cofactor activity; immune complex


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