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Published ahead of print on March 14, 2003, doi:10.1165/rcmb.2002-0132OC
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American Journal of Respiratory Cell and Molecular Biology. Vol. 29, pp. 252-258, 2003
© 2003 American Thoracic Society
DOI: 10.1165/rcmb.2002-0132OC

Effect of CD14 Blockade on Endotoxin-Induced Acute Lung Injury in Mice

Sadatomo Tasaka, Akitoshi Ishizaka, Wakako Yamada, Mie Shimizu, Hidefumi Koh, Naoki Hasegawa, Yoshiyuki Adachi and Kazuhiro Yamaguchi

Department of Medicine, Keio University School of Medicine; Department of Laboratory Medicine, Tokyo Electric Power Company Hospital; and Laboratory of Immunopharmacology of Microbial Products, Tokyo University of Pharmacy and Life Science, Tokyo, Japan

Address correspondence to: Akitoshi Ishizaka, M.D., Department of Laboratory Medicine, Tokyo Electric Power Company Hospital, Shinjuku-ku, Tokyo 160-1100, Japan. E-mail: ishiz{at}attglobal.net

CD14 functions as a cell surface receptor for endotoxin (lipopolysaccharide [LPS]) and is thought to have an essential role in innate immune responses to infection. Previous studies have revealed attenuation of the systemic response after sepsis by blocking CD14. In this study, we tested the hypothesis that CD14 blockade protects against inflammatory responses associated with LPS pneumonia. We examined the effect of an anti-murine CD14 monoclonal antibody (4C1) on the development of acute lung injury induced by intratracheal LPS in mice. We also measured the production of cytokines (tumor necrosis factor-{alpha}, interleukin-6, and macrophage inflammatory protein-2) and nitric oxide by murine peritoneal macrophages exposed to LPS in vitro. Nuclear factor (NF)-{kappa}B translocation was evaluated in nuclear extracts from lung homogenates. 4C1 significantly attenuated pulmonary edema and neutrophil emigration after LPS administration. The production of cytokines and nitric oxide by LPS-stimulated macrophages was significantly decreased by 4C1 treatment. NF-{kappa}B translocation induced by LPS instillation was also suppressed by 4C1. These results suggest that blockade of CD14 might attenuate acute lung injury after intratracheal instillation of LPS through the suppression of NF-{kappa}B translocation. The inhibitory effect of CD14 blockade on cytokine production and nitric oxide release of macrophages might contribute to the attenuation of lung injury.

Abbreviations: acute respiratory distress syndrome, ARDS • bronchoalveolar lavage, BAL • PBS-T containing 0.5% bovine serum albumin, BPBS-T • enzyme-linked immunosorbent assay, ELISA • electrophoretic mobility shift assay, EMSA • extravascular albumin, EVA • interleukin, IL • LPS-binding protein, LBP • lipopolysaccharide, LPS • macrophage inflammatory protein, MIP • nuclear factor-{kappa}B, NF-{kappa}B • phosphate-buffered saline, PBS • PBS containing 0.05% Tween 20 • PBS-T • red blood cells, RBC • reticuloendothelial system, RES • tumor necrosis factor-{alpha}, TNF-{alpha} • white blood cells, WBC




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