To generate a recombinant fusion protein (FP) based on the endotoxin-binding domain of bactericidal/permeability-increasing protein (BPI) and the constant domain of IgG and to test its ability to inhibit lipopolysaccharide (LPS)-induced macrophage tumor necrosis factor α (TNF-α) secretion.
A murine macrophage cell line, RAW 264.7, was transfected with a BPI-IgG FP before incubation with LPS. The amount of LPS-induced TNF-α protein secreted was measured and compared with that secreted by cells transfected with a control construct.
Basic science research laboratory.
Main Outcome Measure:
Secreted TNF-α protein concentration.
After transfection, RAW 264.7–cell FP expression was detected in cell lysates and supernatants. At each LPS dose tested, cells transfected with the FP gene secreted less TNF-α than did cells transfected with a control construct.
The FP possesses substantial antiendotoxin activity, as delineated by inhibition of LPS-induced TNF-α secretion by murine macrophages transfected with the fusion gene construct. In the future, such FP may be used as a clinical reagent to reduce the morbidity and mortality associated with serious gram-negative bacterial infections in surgical patients.Arch Surg. 1996;131:1173-1178
Dahlberg PS, Acton RD, Uknis ME, Klaerner HG, Johnston JW, Levelle CD, Gray BH, Dunn DL. Macrophages Expressing a Fusion Protein Derived From Bactericidal/Permeability-Increasing Protein and IgG Are Resistant to Endotoxin. Arch Surg. 1996;131(11):1173-1178. doi:10.1001/archsurg.1996.01430230055010