Detection of tumor necrosis factor soluble receptor p55 in blood samples from healthy and endotoxemic humans.
The tumor necrosis factor (TNF) soluble receptor derived from the cell surface p55 TNF receptor (TNFsRp55) is a naturally occurring substance generated during infection and inflammation. TNFsRp55 inhibits biologic effects of TNF. An RIA was developed to quantitate TNFsRp55 in human blood. Recovery of TNFsRp55 from blood anticoagulated with EDTA was optimal compared with recovery from serum or heparinized plasma. TNF did not interfere with the assay. With the RIA based on radiolabeled nonglycosylated Escherichia coli-derived recombinant TNFsRp55, a mean concentration of 198 +/- 15 pg/mL was found in 14 volunteers. When glycosylated CHO cell-derived TNFsRp55 was used, the mean level was 1656 +/- 95 pg/mL. Infusion of endotoxin into volunteers induced TNFsRp55, which peaked at 517 +/- 99 pg/mL for the E. coli-based RIA and 7300 +/- 1810 pg/mL for the CHO cell-based RIA. These findings demonstrate that blood collected in EDTA is optimal for measuring circulating TNFsRp55 and that this soluble receptor is present in health but elevated during endotoxemia.
Dept. of Medicine, New England Medical Center, Boston, MA 02111.
L
L
Shapiro
B D
BD
Clark
S F
SF
Orencole
D D
DD
Poutsiaka
E V
EV
Granowitz
C A
CA
Dinarello
D000293:Adolescent; D000328:Adult; D001667:Binding, Competitive; D004731:Endotoxins / Q000097:blood*; D004926:Escherichia coli; D006801:Humans; D008297:Male; D011092:Polyethylene Glycols; D011863:Radioimmunoassay; D011956:Receptors, Cell Surface / Q000032:analysis*; D018124:Receptors, Tumor Necrosis Factor; D015203:Reproducibility of Results; D012995:Solubility; D014115:Toxemia / Q000097:blood* / Q000209:etiology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D004731:Endotoxins; D011956:Receptors, Cell Surface; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha; D011092:Polyethylene Glycols; C044101:endotoxin, Escherichia coli