The glycosaminoglycans in cultures of stimulated rat peritoneal macrophages. 2. Gel chromatographic studies and the behaviour of heparan sulfate.
The molecular weight distribution of pMP-derived glycosaminoglycans (GAG), i.e. non-sulfated GAG, chondroitin sulfate (CS), and heparin sulfate (HS)-like material was determined. The peritoneal macrophages (pMP) were harvested from rats normal or stimulated by i.p. injection of thioglycolate, carrageenan or BCG, and maintained in culture. The GAG of cell layer and medium were isolated separately after labeling with 35S-sulfate and 3H-acetate. Treatment with nitrous acid served to remove HS-like material. Labeling with 3H-acetate served to detect synthesis of the high m. w. hyaluronic acid (HA). Gel chromatic separation was done using Sephadex G-200 columns. The maximal size of 35S-labeled GAG, especially HS (36 kDa), was reduced in cultural medium and cell layer after stimulation in vivo. Reduction was most pronounced after application of carrageenan followed by thioglycolate and BCG/LPS stimulation. The extracellular GAG of BCG-stimulated pMP were smallest, probably due to degradation. Heparan sulfate-like material made up a larger proportion in monolayer and medium, comprising the total m.w. range up to 36 kDa. The GAG sensitive to nitrous acid were maximal in cultures of carrageenan-stimulated pMP and minimal in those of thioglycolate-stimulated pMP. This type of HS was sensitive to hyaluronidase, too. Any synthesis of high molecular hyaluronic acid was not found in normal or stimulated rat pMP. Therefore MP-associated HA must be adsorbed from other sources or synthesized by early forms of macrophages.
Kittlick
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PD
Institute of Pathology, Friedrich-Schiller-University, Jena, Germany.
Engelmann
D
D
D000818:Animals; D001500:BCG Vaccine / Q000494:pharmacology; D002351:Carrageenan / Q000494:pharmacology; D002478:Cells, Cultured; D006025:Glycosaminoglycans / Q000096:biosynthesis / Q000737:chemistry*; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D010529:Peritoneal Cavity / Q000166:cytology*; D051381:Rats; D017208:Rats, Wistar; D013864:Thioglycolates / Q000494:pharmacology
D001500:BCG Vaccine; D006025:Glycosaminoglycans; D013864:Thioglycolates; D002351:Carrageenan