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Enzyme-linked immunoabsorbent assay detection of a soluble form of
urokinase plasminogen activator receptor in vivo
IF Mizukami, NE Faulkner, MR Gyetko, RG Sitrin and RF Todd
Department of Internal Medicine, University of Michigan Medical School, Ann
Arbor, USA.
The receptor for urokinase plasminogen activator (uPA-R, CD87) is a
glycosylphosphatidylinositol (GPI)-anchored 50 to 65 kD glycoprotein that,
by regulating membrane-associated plasmin activity, may facilitate the
invasion of inflammatory and malignant cells. Certain other GPI-anchored
glycoproteins are shed from the cell membrane and exist as soluble products
in vitro and in vivo. To determine if uPA-R undergoes a similar phenomenon,
we have developed a sensitive enzyme- linked immunoabsorbent assay (ELISA)
(using a rabbit antiserum as both capture and detection reagents) to
measure the quantity of soluble uPA- R (suPA-R) in tissue culture
supernatants and biologic fluids. Using this ELISA, we have detected suPA-R
in the culture supernatants of U- 937 cells and human monocytes stimulated
in vitro by certain soluble inflammatory mediators (Sitrin et al, Blood
84:1268, 1994; Mizukami et al., Clin Res 42:115A, 1994). To determine if
suPA-R exists in vivo, we have screened the plasma of 20 normal volunteers
(mean +/- SD, 3 +/- 3 ng/mL; median, 2 ng/mL; range, 1 to 11 ng/mL [serum
values slightly higher]); the plasma of 13 ICU patients with clinical
sepsis syndrome (mean +/- SD, 30 +/- 11 ng/mL; median, 11 ng/mL; range, 4
to 221 ng/mL); and the extravascular fluids (pleural, pericardial, and
peritoneal) of 84 individuals with presumed inflammatory or malignant
conditions (mean +/- SD, 21 +/- 39 ng/mL; median, 10 ng/mL; range, 2 to 253
ng/mL). Among the latter specimens, most were inflammatory exudates (only
six were malignant by positive cytology) with the highest quantities of
suPA-R associated with neutrophilic exudates. The solubility of suPA-R
contained within these fluids was confirmed by reanalysis after
ultracentrifugation to remove particulate material. When tested in a uPA
ligand capture ELISA, representative specimens of extravascular body fluids
and sepsis plasma contained suPA-R capable of binding uPA ligand (generally
representing a small fraction of the immunoreactive material). We conclude
from these data that suPA-R is immunologically detectable in vitro and in
vivo with high concentrations of receptor found under conditions of
inflammatory stimulation. The possibility of suPA-R's biologic activity is
suggested by its partial retention of ligand binding capacity.
Volume 86,
Issue 1,
pp. 203-211,
07/01/1995
Copyright © 1995 by The American Society of Hematology
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