In vitro analysis of pulmonary inflammation using rat lung organ cultures.
We have analyzed leukocyte to lung adhesive interactions and neutrophil-mediated lung injury using a rat lung organ culture system. Rat lung slices were maintained in tissue culture on gelatin sponges floating at the gas-liquid interface. Maintenance of three-dimensional alveolar structure, critical to the viability of lung tissue, was achieved by instilling 0.5% agarose (in 37 degrees C RPMI 1640) into the lungs during tissue explanation. Quantitative neutrophil to lung adhesive interactions were examined using an adaptation of the Woodruff-Stamper frozen section binding assay. Pretreatment of organ cultures with recombinant human tumor necrosis factor (rhTNF) resulted in a protein synthesis-dependent three- to fourfold increase in adhesiveness for neutrophils. Time course and mononuclear leukocyte binding experiments revealed that TNF-induced rat lung adhesiveness peaks at 4 h and is largely neutrophil-specific. Agonist-induced activation of neutrophils in the presence of [3H]leucine-labeled organ cultures resulted in lung injury as assessed by radioisotope release. These observations are consistent with endothelial cell culture data that indicate that TNF-induced endothelium exhibits a protein synthesis-dependent increase in adhesiveness for neutrophils. These data validate rat lung organ cultures as a model system that can be used to assess mechanisms of neutrophil adhesion and leukocyte-mediated tissue injury.
Warren
J S
JS
Department of Pathology, University of Michigan Medical School, Ann Arbor 48109-0602.
Barton
P A
PA
D000818:Animals; D002104:Cadmium / Q000494:pharmacology; D002448:Cell Adhesion / Q000502:physiology; D002469:Cell Separation; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008168:Lung / Q000166:cytology / Q000187:drug effects / Q000473:pathology; D008297:Male; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects; D009924:Organ Culture Techniques; D011014:Pneumonia / Q000473:pathology*; D051381:Rats; D013696:Temperature
D002104:Cadmium