Yusoff P, Hamilton J A, Nolan R D, Phillips W A
University of Melbourne Department of Medicine, Royal Melbourne Hospital, Victoria, Australia.
Growth Factors. 1994;10(3):181-92. doi: 10.3109/08977199409000236.
The activity of phosphatidylinositol (PI) 3-kinase was examined in murine bone marrow-derived macrophages (BMM) stimulated with the haematopoietic growth factors colony stimulating factor-1 (CSF-1) and granulocyte/macrophage-CSF (GM-CSF). PI 3-kinase was immunoprecipitated from cell lysates using anti-phosphotyrosine antibody or an antibody directed against the 85K subunit of PI 3-kinase, and the activity assayed by the phosphorylation of PI in the presence of [gamma 32P]-ATP. The results demonstrate that CSF-1 increases the activity of PI 3-kinase, as compared to the non-stimulated control, in murine macrophages. Maximum activity was seen after 10 min of stimulation with CSF-1 at 3000-5000 U/ml. The dose-response of CSF-1 is consistent with other biochemical effects of CSF-1 seen in the BMM. GM-CSF also stimulated PI 3-kinase activity although to a lesser extent than CSF-1, correlating well with their degree of mitogenic activity on the BMM. Non-mitogenic macrophage activating agents, such as the phorbol myristate acetate, lipopolysaccharide, concanavalin A and formyl-methionyl-leucyl-phenylalanine, did not significantly increase the PI 3-kinase activity. Furthermore, CSF-1 failed to stimulate PI 3-kinase activity in resident peritoneal macrophages, a population of macrophages with poor proliferative capacity. These results suggest that the PI 3-kinase activity may be involved in the haemopoietic growth factor signalling pathways regulating macrophage growth.
在受到造血生长因子集落刺激因子-1（CSF-1）和粒细胞/巨噬细胞集落刺激因子（GM-CSF）刺激的小鼠骨髓来源巨噬细胞（BMM）中检测了磷脂酰肌醇（PI）3激酶的活性。使用抗磷酸酪氨酸抗体或针对PI 3激酶85K亚基的抗体从细胞裂解物中免疫沉淀PI 3激酶，并在[γ-32P] -ATP存在下通过PI的磷酸化测定活性。结果表明，与未刺激的对照相比，CSF-1可增加小鼠巨噬细胞中PI 3激酶的活性。在3000-5000 U/ml的CSF-1刺激10分钟后观察到最大活性。CSF-1的剂量反应与在BMM中观察到的CSF-1的其他生化效应一致。GM-CSF也刺激了PI 3激酶活性，尽管程度低于CSF-1，这与其对BMM的促有丝分裂活性程度密切相关。非促有丝分裂的巨噬细胞激活剂，如佛波酯肉豆蔻酸酯、脂多糖、伴刀豆球蛋白A和甲酰甲硫氨酰亮氨酰苯丙氨酸，并未显著增加PI 3激酶活性。此外，CSF-1未能刺激驻留腹膜巨噬细胞中的PI 3激酶活性，驻留腹膜巨噬细胞是一群增殖能力较差的巨噬细胞。这些结果表明，PI 3激酶活性可能参与调节巨噬细胞生长的造血生长因子信号通路。
