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Conversion of Normal Ly-1-Positive B-Lineage Cells into Ly-1-Positive Macrophages in Long-Term Bone Marrow Cultures

机译:在长期的骨髓培养中将正常的Ly-1阳性B谱系细胞转换为Ly-1阳性巨噬细胞。

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摘要

We obtained eight different cell lines in the long-term bone marrow culture system thatshowed a germ-line configuration of the joining (J) region segments of the Ig heavy-chain(IgH) genes. Their surface markers were CD45R+, Ly-1+, Lyb-2+, cIgM-, sIgM-, Ia-, Thy-1-,Mac-1-, and IL-2R (Tac)+. Use of very young mice and the presence of IL-5 were important for preferential promotion of the survival of B-lineage lymphocytes bearing the Ly-1markers. When we treated two of them (J8 and J10) with 5-azacytidine for 24 h followed byco-culture with stromal cells and IL-.5, they became Ly-1+, sIgM+ B cells, and Ly-1+, Mac-1+macrophagelike cells, respectively. After other early lymphoid lines (J1, J8, and J13) weremaintained by co-culture with ST2 and IL-5 for more than a year, they showed a heterogeneousDNA rearrangement profile of the J region segment of the IgH gene, although onlyJ13 rearranged the κ-light chain gene. Northern blot analysis revealed that these cell linesexpressed Cμ-mRNA, and λ5-mRNA, consistent with normal pre-B cells. Intriguingly, J1, J8,and J13 expressed c-fms mRNA constitutively. When J13 cells were co-cultured with ST2 andGM-CSF in place of ST2 and IL-5, they acquired Mac-1 expression and retained Ly-1expression. They were morphologically macrophages, nonspecific-esterase-positive, andshowed phagocytosis of latex beads. These results support evidence for a close relationshipbetween the myeloid and Ly-1+ B-cell pathways of differentiation, and indicate that our IL-5-dependent clones are multipotential intermediates in differentiation from pro-B cells to Bcells and macrophages.
机译:我们在长期的骨髓培养系统中获得了八种不同的细胞系,这些细胞系显示了Ig重链(IgH)基因的连接(J)区域片段的种系构型。它们的表面标记是CD45R +,Ly-1 +,Lyb-2 +,cIgM-,sIgM-,Ia-,Thy-1-,Mac-1-和IL-2R(Tac)+。幼鼠的使用和IL-5的存在对于优先促进带有Ly-1标记的B谱系淋巴细胞的存活很重要。当我们将其中两个(J8和J10)用5-氮杂胞苷处理24小时,然后与基质细胞和IL-.5共培养时,它们变成Ly-1 +,sIgM + B细胞和Ly-1 +,Mac- 1+巨噬细胞样细胞。与ST2和IL-5共培养保持其他早期淋巴样细胞系(J1,J8和J13)一年以上后,尽管只有J13重新排列了IgH基因的J区片段,但它们显示了异质的DNA重排图谱。 κ轻链基因。 Northern印迹分析显示,这些细胞系表达Cμ-mRNA和λ5-mRNA,与正常的B前细胞一致。有趣的是,J1,J8和J13组成性表达c-fms mRNA。当J13细胞与ST2和GM-CSF代替ST2和IL-5共培养时,它们获得了Mac-1表达并保留了Ly-1表达。它们在形态上是巨噬细胞,非特异性酯酶阳性,并显示出乳胶珠的吞噬作用。这些结果支持了骨髓和Ly-1 + B细胞分化途径之间密切关系的证据,并表明我们的IL-5依赖性克隆是从pro B细胞向B细胞和巨噬细胞分化的多能中间产物。

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