Cystatin C and cathepsin B production by alveolar macrophages from smokers and nonsmokers.
The capacity of alveolar macrophages (AM) obtained from smokers and nonsmokers to secrete cathepsin B and its inhibitor cystatin C was examined because of the concept that an imbalance in the production of proteolytic enzymes and/or their inhibitors could be responsible for the lung damage seen in smokers. Quantitation of immunoprecipitates on Western blots showed that the amount of total cystatin C secreted into the culture medium by AM of smokers was significantly greater than the amount secreted by cells obtained from nonsmokers, whereas the difference between the amount of cathepsin B secreted by the AM of smokers and that from nonsmokers did not appear significant. The cystatin C found in the medium conditioned by AM of nonsmokers appeared to be more heterogeneous in molecular size, presenting either as a single band of about 14 Kd or as a high-molecular-weight triplet of about 69 Kd, 63 Kd, and 57.3 Kd. Furthermore, in some cases there were single or doublet bands at 14 Kd as well as the high-molecular-weight triplets. In contrast, smokers AM-conditioned medium uniformly possessed both the low-and the high-molecular-weight cystatin C. Cathepsin B was not detected in Western blots at its reported molecular weights but was identified at the exact area occupied by the higher molecular weight cystatin C, i.e., at bands corresponding to 69 Kd, 63 Kd, and 57.3 Kd. Therefore, it is clear that in culture media of AM, cystatin C and cathepsin B are present as proteinase-antiproteinase complexes. The observation also suggests that in smokers an excess of cystatin C may be elaborated, which, if further substantiated, would show for the first time a likely role for this proteinase inhibitor in vivo.
Warfel
A H
AH
Department of Medicine, New York University Medical Center, NY 10016.
Cardozo
C
C
Yoo
O H
OH
Zucker-Franklin
D
D
D000328:Adult; D002401:Cathepsin B / Q000096:biosynthesis*; D002478:Cells, Cultured; D055316:Cystatin C; D015891:Cystatins / Q000096:biosynthesis*; D005260:Female; D006801:Humans; D008264:Macrophages / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D008970:Molecular Weight; D011650:Pulmonary Alveoli / Q000378:metabolism*; D012907:Smoking / Q000378:metabolism*
C525030:CST3 protein, human; D055316:Cystatin C; D015891:Cystatins; D002401:Cathepsin B