HL-60 cells degrade alpha-actinin to produce a fragment that promotes monocyte/macrophage maturation.
An amino-terminal fragment of alpha-actinin can promote monocyte/macrophage maturation. This fragment was initially isolated from media of HL-60 myeloid leukemia cells cultured on extracellular bone marrow matrix. To determine the source of this fragment in this culture system, we investigated whether HL-60 cells grown on bone marrow stroma have increased intracellular levels of alpha-actinin that may be released into the media during cell apoptosis. HL-60 cells grown on matrix showed no evidence of increased cellular alpha-actinin compared to cells grown on plastic substrata as measured by flow cytometry. In addition, there was no evidence of increased apoptosis as determined by DNA fragmentation assays or flow cytometry. However, 100 kD alpha-actinin was found in the extracellular matrix of bone marrow stroma by Western blot analysis and immunofluorescence microscopy. The alpha-actinin content in the stroma was markedly decreased after exposure to HL-60 cells. Furthermore, lysates of HL-60 cells or of peripheral blood monocytes can degrade exogenous alpha-actinin to produce a 31 kD fragment, which promotes monocyte/macrophage maturation. We conclude that when alpha-actinin is present in the extracellular matrix, it can be modified by HL-60 cells to produce a maturation promoting 31 kD fragment.
Masri
M
M
Veterans Affairs Medical Center, Minneapolis, MN 55417, USA.
Wahl
D
D
Oegema
T
T
Luikart
S
S
D000185:Actinin / Q000378:metabolism* / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D005109:Extracellular Matrix / Q000378:metabolism*; D005434:Flow Cytometry; D018922:HL-60 Cells / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008264:Macrophages / Q000166:cytology; D009000:Monocytes / Q000166:cytology; D010446:Peptide Fragments / Q000378:metabolism* / Q000494:pharmacology
D010446:Peptide Fragments; D000185:Actinin