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1 Institute for Environmental Medicine, University of Pennsylvania Medical Center, Philadelphia, Pennsylvania 19104-6068; and 2 Department of Cell Biology, Utrecht University Medical School, 3584 CX Utrecht, The Netherlands
Monoclonal antibodies against the limiting
membrane of alveolar type II cell lamellar bodies were obtained after
immunization of mice with a membrane fraction prepared from lamellar
bodies isolated from rat lungs. The specificity of the antibodies was investigated with Western blot analysis, indirect immunofluorescence, and electron-microscopic immunogold studies of freshly isolated or
cultured alveolar type II cells, alveolar macrophages, and rat lung
tissue. One of the monoclonal antibodies identified, MAb 3C9,
recognized a 180-kDa lamellar body membrane (lbm180) protein.
Immunogold labeling of rat lung tissue with MAb 3C9 demonstrated that
lbm180 protein is primarily localized at the lamellar body limiting
membrane and is not found in the lamellar body contents. Most
multivesicular bodies of type II cells were also labeled, as were some
small cytoplasmic vesicles. Golgi complex labeling and plasma membrane
labeling were weak. The appearance of lbm180 protein by
immunofluorescence in fetal rat lung cryosections correlated with the
biogenesis of lamellar bodies. The lbm180 protein decreased with time
in type II cells cultured on plastic. The lbm180 protein is an integral
membrane protein of lamellar bodies and was also found in the pancreas
and the pancreatic 
HC9 cell line but not in the rat brain, liver,
kidney, stomach, or intestine. The present study provides evidence that
the lbm180 protein is a lung lamellar body and/or
multivesicular body membrane protein and that its antibody, MAb 3C9,
will be a valuable reagent in further investigations of the biogenesis
and trafficking of type II cell organelles.
surfactant; multivesicular bodies; pancreatic -cell
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