Molecular and functional properties of myeloid stem cell growth factors

1)大鼠脾细胞系RSP-2.P3中中性粒细胞生长因子(NeuGF)的纯化及N端氨基酸序列测定。2)正常人尿液中巨噬细胞生长因子(M&lt；Phi&Gt；GF)的纯化和一级结构。我们纯化的M&lt；Phi&Gt；GF的氨基酸末端序列与其他人的一致，但M&lt；Phi&Gt；GF制剂的氨基酸组成表明它来自较大尺寸的M&lt；Phi&Gt；GFmRNA，并经190.3)位附近的加工修饰，该克隆内异质性同时分泌NeuGF和M&lt；Phi&GT；GF。建立了一个新的细胞系，并进行了亚克隆。分离后不久，观察到这些亚克隆之间在产生NeuGF方面与M&lt；Phi&gt；GF不同。然而，在培养一段时间后，这些亚克隆变得彼此相似。4)纯化的NeuGF和M&lt；Phi&Gt；GF的生物活性。骨髓干细胞对NEGF和M&lt；Phi&Gt；GF均有反应，M&lt；Phi&Gt；GF输注6d后小鼠巨噬祖细胞增多，M&lt；Phi&Gt；GF等对腹膜滞留巨噬细胞有激活作用。

1) Purification and N-terminal Amino Acid Sequence of Neutrophil Growth Factor (NeuGF) from Rat Spleen Cell Line RSP-2.P3.Production of NeuGF was optimized by adding inducers to the cell culture, and the secreted NeuGF was purified by reverse-phase HPLC. Elucidated N-terminal sequence of the NeuGF was consistent with that deduced by others by recombinant DNA technology.2) Purification and Primary Structure of Macrophage Growth Factor (M<phi>GF) from Normal Human Urine. Amino terminal sequence of the M<phi>GF we purified was consistent with that deduced by others by sequenceing of its cDNA, but amino acid composition of the M<phi>GF preparation suggested that it is originated from the larger-size M<phi>GF mRNA and modified by processing around the position 190.3) Intraclonal Heterogeneity of Mouse Fibrosarcoma Cells That Secrete Both NeuGF and M<phi>GF. A new cell line was established and subcloned. Soon after isolation, dissimilarity was observed among these subclones regarding the production of NeuGF relative to M<phi>GF. However, the subclones became similar to each other when kept in culture for some time. The result suggests that equilibrated diversity is maintained as a whole population although individual cells dynamically fluctuate.4) Biological Activity of the Purified NeuGF and M<phi>GF. Presence of bone marrow stem cells reactive both to NeGF and to M<phi>GF, increase of macrophage progenitor cells after 6-day's infusion of M<phi>GF in mice, activation of peritoneal resident macrophages by M<phi>GF etc. were demonstrated.