Lipopolysaccharide binding protein and CD14 interaction induces tumor necrosis factor-alpha generation and neutrophil sequestration in lungs after intratracheal endotoxin.
It has been proposed that lipopolysaccharide (LPS) bound to the 60-kD LPS binding protein (LBP) forms an LPS/LBP complex that, in turn, binds to the CD14 receptor on monocytes/macrophages and stimulates the release of cytokines. We examined the role of LBP and CD14 in tumor necrosis factor-alpha (TNF-alpha) production and neutrophil (polymorphonuclear leukocyte [PMN]) sequestration in lungs induced by intratracheal instillation of LPS using rabbit lungs perfused at constant flow with lactated Ringer-albumin solution. LPS alone (Salmonella minnesota, wild type; 20 ng) or in the presence of LBP (500 ng) was injected intratracheally. In some experiments, human PMNs (5 x 10(7)) were added to the perfusate after a 2-hour period of perfusion. Samples of lung perfusate were collected every 30 minutes for 180 minutes when bronchoalveolar lavage was also performed. TNF-alpha concentrations in the perfusate and bronchoalveolar lavage fluid were determined by use of a bioassay with L-929 fibroblasts, and PMN accumulation in lungs was determined by myeloperoxidase assay of lung homogenates. LPS alone did not significantly increase TNF-alpha production or lung PMN accumulation, whereas the LPS/LBP complex increased TNF-alpha concentration in perfusate twofold and PMN accumulation twofold compared with the effect of LPS alone. Intratracheal instillation of anti-CD14 monoclonal antibody MY4 (40 micrograms) with the LPS/LBP complex prevented TNF-alpha release and PMN sequestration, whereas an isotype-matched control monoclonal antibody was ineffective. Therefore, LBP in the airspace enhances the LPS effect on TNF-alpha production via a CD14-dependent pathway, and as a result, CD14 activation can contribute to lung PMN sequestration.(ABSTRACT TRUNCATED AT 250 WORDS)
Ishii
Y
Y
Department of Physiology and Cell Biology, Albany Medical College, NY 12208.
Wang
Y
Y
Haziot
A
A
del Vecchio
P J
PJ
Goyert
S M
SM
Malik
A B
AB
D000209:Acute-Phase Proteins; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000494:pharmacology; D015703:Antigens, CD / Q000378:metabolism / Q000494:pharmacology*; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism / Q000494:pharmacology*; D001992:Bronchoalveolar Lavage Fluid / Q000378:metabolism*; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D004347:Drug Interactions; D006801:Humans; D018950:Lipopolysaccharide Receptors; D008168:Lung / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016676:Macrophages, Alveolar / Q000378:metabolism*; D008562:Membrane Glycoproteins; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D011817:Rabbits; D011946:Receptors, Antigen / Q000032:analysis; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*; D000099231:Lipopolysaccharide-Binding Protein
D000209:Acute-Phase Proteins; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D002352:Carrier Proteins; D018950:Lipopolysaccharide Receptors; D008562:Membrane Glycoproteins; D011946:Receptors, Antigen; D014409:Tumor Necrosis Factor-alpha; D000099231:Lipopolysaccharide-Binding Protein