Acetylated lipoproteins impair erythroid growth factor release from endothelial cells.
Endothelial cells are a known source of hematopoietic growth-enhancing factors, including platelet-derived growth factor (PDGF). In addition, endothelium interacts directly with plasma lipoproteins which have been shown to modulate hematopoiesis. To determine the relationship of these properties, we measured the release of an erythroid growth-enhancing factor from bovine endothelial cells under lipid-loaded and control conditions. Human bone marrow cells cultured under serum-free conditions form more erythroid, granulocyte/macrophage, and mixed hematopoietic colonies when supplemented with endothelial cell-conditioned medium (ECCM) than do controls (P less than 0.05). The activity is expressed over a wide range of erythropoietin, lymphocyte-conditioned medium (LCM), recombinant human interleukin-3, and colony-stimulating factor (CSF) concentrations, and is related to ECCM dose. In contrast, enhancing activity in ECCM prepared with 0-400 micrograms/ml acetylated low density lipoproteins (AcLDL) or native LDL is diminished to 0% in a dose-dependent fashion (relative to ECCM from unexposed cells or from cells incubated with very low density lipoproteins, P less than 0.05). Upon dilution, medium prepared from cells incubated with LDL shows a rightward shift in the dose-response curve for erythroid colony formation, while that prepared from AcLDL loaded cells demonstrates a downward shift, indicating that the inhibitory activities are kinetically distinct. Delipidation of ECCM prior to addition to marrow culture removes the inhibitory action of native LDL (P less than 0.05) but not that of AcLDL (P greater than 0.10). Immunochemical analysis suggests that the erythropoietic activity in ECCM is unrelated to that of PDGF, recombinant human CSF, and erythroid burst-promoting activity (BPA) present in LCM. This conclusion is supported by Northern blot analysis of endothelial cells using a cDNA probe for the v-sis homologue of the PDGF beta chain and by immunoprecipitation of metabolically labeled PDGF. The relative amounts of c-sis transcripts and of secreted PDGF were similar in endothelial cells incubated with or without AcLDL. We conclude that AcLDL impair the synthesis or release of an erythropoietic growth-enhancing factor(s) which is biologically distinct from PDGF and BPA present in LCM.
Dainiak
N
N
Department of Medicine, St. Elizabeth's Hospital, Boston, MA 02135.
Warren
H B
HB
Kreczko
S
S
Riordan
M A
MA
Feldman
L
L
Lawler
J
J
Cohen
A M
AM
Davies
P F
PF
D000107:Acetylation; D000818:Animals; D002417:Cattle; D002784:Cholesterol / Q000494:pharmacology; D003114:Colony-Forming Units Assay; D003470:Culture Media / Q000032:analysis; D004347:Drug Interactions; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004920:Erythropoiesis / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D006133:Growth Substances / Q000378:metabolism* / Q000502:physiology; D016298:Hematopoietic Cell Growth Factors; D006801:Humans; D050356:Lipid Metabolism; D008055:Lipids / Q000494:pharmacology; D008077:Lipoproteins, LDL / Q000378:metabolism*; D010982:Platelet-Derived Growth Factor / Q000235:genetics
D003470:Culture Media; D006133:Growth Substances; D016298:Hematopoietic Cell Growth Factors; D008055:Lipids; D008077:Lipoproteins, LDL; D010982:Platelet-Derived Growth Factor; D002784:Cholesterol