PMID: 8967503May 1, 1996Paper

Endothelial cells internalize monoclonal antibody to angiotensin-converting enzyme

The American Journal of Physiology
V MuzykantovA B Fisher

Abstract

We investigated the fate of MAb 9B9, a monoclonal antibody to angiotensin-converting enzyme (ACE), which binds to endothelium both in vitro and in vivo. Using cultured human umbilical vein endothelial cells (HUVEC) and isolated perfused rat lungs (IPL), we demonstrated specific and saturable binding of 125I-labeled MAb 9B9 at 4 degrees C [affinity constant (Kd) = 20-50 nM, maximal number of binding sites (Bmax) = 1.5-3.0 x 10(5) sites/cell]. When 125I-MAb 9B9 was bound to HUVEC at 37 degrees C, only 40% of cell-associated radioactivity was acid elutable, suggesting antibody internalization. This was confirmed by finding that 1) the amount of MAb 9B9 uptake at 37 degrees C was higher than at 4 degrees C both in HUVEC and IPL; 2) binding of 125I-labeled streptavidin with HUVEC and IPL pretreated with biotinylated MAb 9B9 (b-MAb 9B9) was diminished in a temperature- and time-dependent fashion at 37 degrees C; and 3) b-MAb 9B9 bound to HUVEC at 37 degrees C was found intracellularly by ultrastructural analysis using streptavidin gold. Intracellular 125I-MAb 9B9 was found in microsomal fractions of lung homogenate from IPL and after intravenous (iv) injections in rats. Degradation of internalized MAb 9B9 was minimal, since > 90% of ...Continue Reading

Citations

May 15, 2001·American Journal of Physiology. Lung Cellular and Molecular Physiology·S M DanilovV R Muzykantov
Apr 17, 2007·American Journal of Physiology. Lung Cellular and Molecular Physiology·Kai NowakSergei M Danilov
Mar 16, 2002·Gene Therapy·I V BalyasnikovaS M Danilov
Jan 18, 2006·Treatments in Respiratory Medicine·Melpo Christofidou-Solomidou, Vladimir R Muzykantov
Feb 13, 2002·American Journal of Physiology. Lung Cellular and Molecular Physiology·Juan-Carlos MurcianoVladimir R Muzykantov
Mar 22, 2005·Pulmonary Pharmacology & Therapeutics·Irina V BalyasnikovaSergei M Danilov

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