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S E Alpert

Publications and source records attributed to S E Alpert.

23 records · Page 2Linked to original sources

Macrophage maturation: differences in complement secretion by marrow, monocyte, and tissue macrophages detected with an improved hemolytic plaque assay.

In order to examine one function of mononuclear phagocytes during maturation from bone marrow precursors to tissue macrophages, an improved hemolytic plaque assay for the detection of synthesis of the second (C2) and fourth (C4) components of C by single cells was developed. With this method, production of C2 and C4 was assessed in cell populations derived from bone marrow, blood, lung, peritoneum, and spleen. The proportion of cells producing C2 and C4 in each population varied. Approximately 10% of bone marrow cells produced C4, but not detectable C2 plaque-forming cells (PFC) were detected. Circulating monocytes yielded about 10% PFC each for C2 and C4. The proportion of C2-producing cells in tissue macrophages varied from approximately 2% in bronchoalveolar macrophages to about 45% in peritoneal and splenic macrophage populations, whereas C4 production by macrophages from lung, peritoneum, and spleen were all approximately 45%. These data suggest that differences in C biosynthesis characterize mononuclear phagocytes at different stages of maturation.

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Comparative levels of endogenous myelin basic protein-serum factors (MBP-SFs) in adult and suckling mice (B6CBAF1 and B6C3HF1, strains) and in neurologically mutant mice of the same genetic background.

Normal adult B6C3HF1 and B6CBAF1 mice contained at least 10 times higher levels (1.17 microM) of endogenous myelin basic protein-serum factors (MBP-SFs) than previously found in adult Lewis rats. In rat MBP-SF levels in the adult (0.03 microM) were much less than in the suckling animals (0.74 microM). In mice, by contrast, the adult (1.17 microM) and suckling (0.75 microM) levels were similar. Suckling mice from 9 different neurologically mutant strains and their clinically normal littermates had MBP-SF levels (0.5 microM) slightly below that of normal suckling mice of the same genetic background (0.75 microM).

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Generation of lipoxygenase metabolites of arachidonic acid by monolayer cultures of tracheal epithelial cells and intact tracheal segments from rabbits.

We compared the profile of lipoxygenase metabolites of arachidonic acid (AA) generated by cultured rabbit tracheal epithelial (TE) cells with that produced by intact rabbit tracheal segments at baseline and following addition of exogenous AA or calcium ionophore A23187. Lipoxygenase metabolites in effluent media were resolved by high-pressure liquid chromatography and quantitated by radioimmunoassay for monohydroxyeicosanoid (HETE) and leukotriene (LT) metabolites [5-, 12-, and 15-HETE; LTB4, LTC4, LTD4]. Following incubation with exogenous AA (10 micrograms/ml), cultured TE cells generated immunoreactive products that coeluted with authentic 5-, 12-, and 15-HETE standards. 12-HETE was the predominant metabolite. Whereas the generation of HETEs by TE monolayers was dependent on addition of exogenous AA, intact tracheal segments demonstrated a baseline production of 12-HETE and lesser amounts of 5- and 15-HETE as well as unidentified metabolites with UV absorbance at 280 nm. Incubation of tracheal segments with AA resulted in augmented metabolite production. In cultured TE cells, small quantities of HETEs were present intracellularly esterified to membrane phospholipids or free in the cytosol, and significant increases in free cytosolic 12- and 15-HETE were detected postincubation with AA. Calcium ionophore (5 microM) did not induce significant increases in HETE production in either cultured TE cells or tracheal segments. Minimal or no immunoreactive LTs B4, C4, and D4 were produced by TE monolayers or tracheal segments at baseline or following addition of AA or ionophore. Production of HETEs by cultured TE cells was not associated with decreased viability, release of intracellular lactic dehydrogenase, or loss of cells from the monolayers. Preincubation of monolayer cultures or tracheal segments with 5,8,11,14-eicosatetraynoic acid prior to addition of exogenous AA inhibition metabolite production. Our observations provide further documentation for the generation of lipoxygenase metabolites by TE cells and suggest that the array of metabolites generated by cultured TE cells may not be representative of the entire spectrum of AA metabolites produced by intact native epithelium.

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Functional consequences of abnormal fatty acid profiles in cultured airway epithelial cells.

Maintenance of serum-free conditions for the culture of TE or other airway epithelial cells provides a defined environment in which to explore the regulation of cellular functions. Yet TE cells appear to be dependent on the medium for essential, if not all, polyunsaturated fatty acids. At present, some laboratories routinely use serum to support the growth of airway epithelial cells, presumably in part through recognition that cells of mammalian origin require an exogenous source of lipids. While 5% FBS can increase the linoleic and arachidonic acid content of cultured rabbit and human TE cells, it does not fully restore the fatty acid composition of cultured TE cells to that of freshly isolated cells, particularly in the case of human TE cells. Equally good, if not better, repair of membrane fatty acid composition can be achieved by addition of a defined, commercial non-serum source of lipids (Excyte III) plus exogenous arachidonic acid. Cultured TE cells maintained in serum-free medium have been shown to be deficient in prostaglandin and HETE production, both at baseline and in response to physiological stimuli compared to TE cells with greater endogenous content of arachidonic acid. Differences between lipid supplemented and unsupplemented cultured TE cells in cAMP response to PGE2 and in susceptibility to hyperoxic injury have been observed. Other cellular functions regulated by the fatty acid composition of membrane lipids may also be impaired in lipid unsupplemented cells. It is evident that the maintenance of as normal as possible membrane fatty acid content is essential to the use of cultured TE cells as experimental models of airway epithelium.

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