Macrophages Expressing a Fusion Protein Derived From Bactericidal/Permeability-Increasing Protein and IgG Are Resistant to Endotoxin
Peter S. Dahlberg, MD;
Robert D. Acton, MD;
Marc E. Uknis, MD;
Hanz G. Klaerner, MD;
Jennifer W. Johnston;
Christopher D. Levelle;
Beulah H. Gray, PhD;
David L. Dunn, MD, PhD
Arch Surg. 1996;131(11):1173-1178.
Abstract
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Objectives
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  (TNF-) secretion.
Design
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- protein secreted was measured and compared with that secreted by cells transfected with a control construct.
Setting
Basic science research laboratory.
Main Outcome Measure
Secreted TNF- protein concentration.
Results
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- than did cells transfected with a control construct.
Conclusions
The FP possesses substantial antiendotoxin activity, as delineated by inhibition of LPS-induced TNF- 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
Author Affiliations
From the Departments of Surgery (Drs Dahlberg, Acton, Uknis, Klaerner, and Dunn and Ms Johnston and Mr Levelle) and Microbiology (Dr Gray), University of Minnesota, Minneapolis.
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