Brasel K, Escobar S, Anderberg R, de Vries P, Gruss H J, Lyman S D
Department of Immunobiology, Immunex Research and Development Corporation, Seattle, Washington 98101, USA.
Leukemia. 1995 Jul;9(7):1212-8.
Expression of the flt3 tyrosine kinase receptor and its ligand were examined on various murine and human hematopoietic cell lines. Surface expression of flt3 receptor and flt3 ligand were detected by flow cytometry using biotinylated human flt3 ligand or biotinylated soluble human flt3 receptor Fc fusion protein (flt3R-Fc), respectively. Flt3 receptor and ligand expression were also examined by Northern blot analysis. Flt3 receptor was expressed on the surface of only two of nine murine cell lines and nine of 15 human cell lines, with positive cells representing the B cell, early myeloid, and monocytic lineages. Staining for surface expression of the flt3 ligand revealed that seven of nine murine cell lines and nine of 15 human cell lines screened were positive by flow cytometry. All murine and human cell lines assayed were positive for flt3 ligand RNA expression by Northern blot analysis, but not all cell lines expressing flt3 ligand mRNA had detectable surface expression. Cells expressing the flt3 ligand were of the myeloid, B cell and T cell lineages at various stages of differentiation. Only the OCI-AML-5, NALM-6, and AML-193 cell lines coexpressed both surface flt3 receptor and ligand. The myeloid leukemic M1 cell terminally differentiate into macrophage-like cells under the influence of leukemia inhibitory factor (LIF). We found that LIF-stimulated M1 cells down-regulated surface expression and mRNA levels of the flt3 receptor, but up-regulated expression of the flt3 ligand. Although we could demonstrate that the flt3 receptor was functional in the M1 cell line, flt3 ligand could not induce the M1 cells to differentiate.
在多种小鼠和人类造血细胞系中检测了flt3酪氨酸激酶受体及其配体的表达。分别使用生物素化的人flt3配体或生物素化的可溶性人flt3受体Fc融合蛋白(flt3R-Fc),通过流式细胞术检测flt3受体和flt3配体的表面表达。还通过Northern印迹分析检测了Flt3受体和配体的表达。Flt3受体仅在9种小鼠细胞系中的2种以及15种人类细胞系中的9种表面表达,阳性细胞代表B细胞、早期髓系和单核细胞系。对flt3配体表面表达的染色显示,通过流式细胞术筛选的9种小鼠细胞系中的7种以及15种人类细胞系中的9种呈阳性。通过Northern印迹分析,所有检测的小鼠和人类细胞系flt3配体RNA表达均为阳性,但并非所有表达flt3配体mRNA的细胞系都有可检测到的表面表达。表达flt3配体的细胞处于不同分化阶段的髓系、B细胞和T细胞系。只有OCI-AML-5、NALM-6和AML-193细胞系同时共表达表面flt3受体和配体。髓系白血病M1细胞在白血病抑制因子(LIF)的影响下终末分化为巨噬细胞样细胞。我们发现LIF刺激的M1细胞下调flt3受体的表面表达和mRNA水平,但上调flt3配体的表达。尽管我们可以证明flt3受体在M1细胞系中具有功能,但flt3配体不能诱导M1细胞分化。