ISOLATION AND CHARACTERIZATION OF A MACROPHAGE-DERIVED HEPARIN-BINDING GROWTH-FACTOR

被引:172
作者
BESNER, G
HIGASHIYAMA, S
KLAGSBRUN, M
机构
[1] CHILDRENS HOSP MED CTR,DEPT SURG,BOSTON,MA 02115
[2] CHILDRENS HOSP MED CTR,DEPT BIOL CHEM,BOSTON,MA 02115
[3] CHILDRENS HOSP MED CTR,DEPT MOLEC PHARMACOL,BOSTON,MA 02115
[4] HARVARD UNIV,SCH MED,BOSTON,MA 02115
来源
CELL REGULATION | 1990年 / 1卷 / 11期
关键词
D O I
10.1091/mbc.1.11.811
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Human mononuclear cells were plated in culture, and the conditioned media of these cells were analyzed by heparin-Sepharose affinity chromatography. The fractions were tested for growth factor activity as measured by the stimulation of DNA synthesis in BALB/c 3T3 cells. After 2 d in culture, two peaks of heparin-binding growth factor (HBGF) activity were detected, one eluting with 0.5 M NaCl, which could be shown to be platelet-derived growth factor (PDGF)-like, and the other eluting with 1.0 M NaCl. After 7-11 d in culture, when monocytes had clearly differentiated into macrophages, >95% of the HBGF activity in conditioned medium consisted of the 1.0 M NaCl elution peak. This activity, which was designated macrophage-derived HBGF (MD-HBGF), was found to be a cationic heat-resistant polypeptide with a molecular weight in the range of 14-25 kDa. Analysis using Western blots and specific neutralizing antisera, as well as comparative heparin affinity analysis, indicated that MD-HBGF was not identical to other heparin-binding 3T3 cell growth factors known to be produced by macrophages, such as PDGF (AB, AA, and BB forms), acidic fibroblast growth factor, and basic fibroblast growth factor. In addition to stimulating mitogenesis in 3T3 cells, MD-HBGF also stimulated the proliferation of vascular smooth muscle cells, but did not stimulate the proliferation of vascular endothelial cells. © 1990 by The American Society for Cell Biology.
引用
收藏
页码:811 / 819
页数:9
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