PRODUCTION OF HUMAN INTERLEUKIN FOR DA CELLS (HILDA) LEUKEMIA INHIBITORY FACTOR (LIF) BY ACTIVATED MONOCYTES
PRODUCTION OF HUMAN INTERLEUKIN FOR DA CELLS (HILDA) LEUKEMIA INHIBITORY FACTOR (LIF) BY ACTIVATED MONOCYTES
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DOI：
10.1016/0008-8749(90)90161-j
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发表时间：
1990-10-01
影响因子：
4.3
通讯作者：
SOULILLOU, JP
中科院分区：
文献类型：
--
作者：
ANEGON, I;MOREAU, JF;SOULILLOU, JP
我们证明了mRNA的积累和生产HILDA/LIF的人活化钼，单核细胞衍生的巨噬细胞和骨髓单核细胞系。在测试的各种刺激物中，佛波醇二酯和VD 3的协同组合是HILDA/LIF基因表达的最有效诱导物。激活钼上的mRNA积累动力学表现出刺激峰在24小时，此后下降。在培养24小时时检测到HILDA/LIF活性培养上清液，并在培养72小时时达到平台。与Mo相比，PBL在与PDBu和VD 3培养时不积累HILDA/LIF mRNA，而PHA以及PDBu和A23187的组合诱导培养上清液中HILDA/LIF mRNA的积累和分泌。为了排除HILDA/LIF是由污染PBL产生的可能性，使用高度富集的Mo制剂，其缺乏T细胞，如通过不存在TCR-β所评估的。链mRNA转录物。刺激的U937细胞条件培养基与125 I标记的nHILDA/LIF竞争结合其在鼠M1细胞上的受体的能力进一步证明了单核细胞产生HILDA/LIF。在PDBu和VD 3刺激的协同作用下，Mo衍生的巨噬细胞以及HL-60和U937细胞系积累HILDA/LIF mRNA，并以与Mo相同的动力学产生这种细胞因子。最后，我们表明，HILDA/LIF mRNA的积累在U937细胞刺激PDBu，或PDBu和VD 3的组合，抑制蛋白质合成抑制剂CHX的存在下。这些结果首次证明，当在体外适当刺激时，人Mo可以表达HILDA/LIF基因及其产物，并且在此过程中必须新合成中间蛋白。
We demonstrate mRNA accumulation and production of HILDA/LIF by human activated Mo, monocyte-derived macrophages and myelomonocytic cell lines. Among the various stimuli tested, the synergistic combination of phorbol diester and VD3 was the most potent inducer of HILDA/LIF gene expression. The kinetics of mRNA accumulation on activated Mo showed a stimulation peak at 24 hr which declined thereafter. HILDA/LIF activity culture supernatants was detected at 24 hr and reached a plateau at 72 hr of culture. IN contrast to Mo, PBL did not accumulate HILDA/LIF mRNA upon culture with PDBu and VD3, whereas PHA and the combination of PDBu and A23187 induced HILDA/LIF mRNA accumulation and secretion in the culture supernatant. To exclude the possibility that HILDA/LIF was produced by contaminating PBL, highly enriched Mo preparations were used, which were devoid of T cells as assessed by the absence of TCR-.beta. chain mRNA transcripts. HILDA/LIF production by monocytic cells was further documented by the capacity of stimulated U937 cell conditioned medium to compete with 125I-labeled nHILDA/LIF for binding to its receptor on murine M1 cells. Under the synergistic effect of PDBu and VD3 stimulation, Mo-derived macrophages as well as HL-60 and U937 cell lines accumulated HILDA/LIF mRNA and produced this cytokine with identical kinetics as for Mo. Finally, we show that HILDA/LIF mRNA accumulation in U937 cells upon stimulation with PDBu, or the combination of PDBu and VD3, was inhibited in the presence of the protein synthesis inhibitor CHX. These results document for the first time that human Mo, when stimulated appropriately in vitro, can express the HILDA/LIF gene and its product, and that intermediate proteins must be newly synthesized in this process.