In vivo detection of a novel macrophage-derived protein involved in the regulation of mucus-like glycoconjugate secretion.
We previously described a novel 68,000 D macrophage-derived protein (MMS-68) that can stimulate mucus-like glycoconjugate (MLGC) secretion from cultured human airways, respiratory epithelial cells, and the ishikawa adenocarcinoma cell line. To better characterize this mucus secretagogue, we generated monoclonal antibodies against MMS-68 by injecting crushed SDS-PAGE gel slices containing this protein into Balb-C mice followed by fusion with SP2/0, a nonsecreting mouse myeloma cell line. A panel of monoclonal antibodies was produced that identified the 68,000 D MMS by immunoblot analysis and immunoprecipitation. The monoclonal antibodies detected MMS-68 in normal peripheral blood monocytes and pulmonary macrophages by cytofluorographic analysis and in human airways as determined by immunohistochemistry. Utilizing the monoclonal antibodies, an antigen-capture ELISA assay was developed. Statistically significant elevations in levels of MMS-68 were detected in bronchoalveolar lavage fluid (BALF) of chronic bronchitic subjects and cigarette smokers and in monocyte culture supernatants from steroid-dependent asthmatic patients compared to normal control subjects. The 68,000 D MMS is a potent secretagogue and may play an important role in the regulation of mucus secretion, especially in chronic bronchitis and steroid-dependent asthma.
Sperber
K
K
Department of Medicine, Mount Sinai Medical Center, New York, New York.
Gollub
E
E
Goswami
S
S
Kalb
T H
TH
Mayer
L
L
Marom
Z
Z
D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal; D001249:Asthma / Q000378:metabolism; D001991:Bronchitis / Q000378:metabolism; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D003593:Cytoplasm / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005455:Fluorescent Antibody Technique; D006001:Glycoconjugates / Q000378:metabolism*; D006168:Guinea Pigs; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007239:Infections / Q000378:metabolism; D016676:Macrophages, Alveolar / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008875:Middle Aged; D009000:Monocytes / Q000737:chemistry; D009093:Mucus / Q000378:metabolism*; D011233:Precipitin Tests; D012907:Smoking
D000911:Antibodies, Monoclonal; D006001:Glycoconjugates