Thrombin binds to murine bone marrow-derived macrophages and enhances colony-stimulating factor-1-driven mitogenesis.
The binding and mitogenic properties of thrombin have been established in various transformed cell lines. In such systems, thrombin induces cell division in the absence of exogenous growth factors, and the enzyme is considered to act directly as a mitogen. This study explores thrombin's interaction with nontransformed, growth factor-dependent cells. Binding of 125I-alpha-thrombin to colony-stimulating factor (CSF)-1-dependent bone marrow-derived macrophages is saturable, time-dependent, and displaceable by both unlabeled alpha-thrombin, and esterolytically inactive thrombin. Both dissociation studies of pre-bound radio-labeled thrombin and Scatchard analysis assisted by the program "Ligand" suggest adherence of thrombin-binding data to a multi-site model. There are an estimated 2 x 10(4) high affinity sites (Kd = 7 x 10(-9)M) and 2 x 10(6) low affinity sites (Kd = 9 x 10(-7)M) per cell. Quiescent bone marrow-derived macrophages were cultured with either 10(-8)M thrombin, 1000 units of CSF-1/ml, or both and [3H]thymidine incorporation was determined. Thrombin alone did not induce mitogenesis. CSF-1 induced mitogenesis with peak [3H] thymidine incorporation occurring 24 h after addition of the mitogen. This CSF-1-dependent mitogenic influence was enhanced greater than 2-fold by treatment with thrombin.
Department of Pathology, Jewish Hospital, Washington University Medical Center, St. Louis, Missouri 63110.
D R
DR
Clohisy
J M
JM
Erdmann
G D
GD
Wilner
D000818:Animals; D001854:Bone Marrow Cells; D002455:Cell Division / Q000187:drug effects; D003115:Colony-Stimulating Factors / Q000494:pharmacology*; D004347:Drug Interactions; D007457:Iodine Radioisotopes; D007700:Kinetics; D016173:Macrophage Colony-Stimulating Factor; D008264:Macrophages / Q000378:metabolism*; D008297:Male; D051379:Mice; D008805:Mice, Inbred A; D008934:Mitogens; D013917:Thrombin / Q000378:metabolism* / Q000494:pharmacology
D003115:Colony-Stimulating Factors; D007457:Iodine Radioisotopes; D008934:Mitogens; D016173:Macrophage Colony-Stimulating Factor; D013917:Thrombin