In situ hybridization for interleukin 2 and interleukin 2 receptor mRNA in T cells activated in the presence or absence of cyclosporin A.
In situ hybridization for interleukin 2 and interleukin 2 receptor mRNA in T cells activated in the presence or absence of cyclosporin A.
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在存在或不存在环孢菌素 A 的情况下激活的 T 细胞中白细胞介素 2 和白细胞介素 2 受体 mRNA 的原位杂交。
DOI：
10.1084/jem.168.5.1649
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发表时间：
1988
期刊：
影响因子：
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通讯作者：
Granelli-Piperno,A
中科院分区：
文献类型：
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作者：
Granelli-Piperno,A
用RNA-RNA原位杂交法研究抗CD3、抗CD28或PHA三种有丝分裂原刺激的T淋巴细胞产生IL-2和IL-2-R mRNA的频率。当这些有丝分裂原加PMA刺激时，CD4+和CD8+T细胞都表达IL-2和低亲和力IL-2-R的转录本。经PHA或抗CD3刺激后，环孢素A(CsA)抑制IL-2的合成，但不抑制IL-2-R的表达。然而，在抗CD28刺激后，需要较高浓度的CsA才能达到50-70%的抑制。在最佳时间点，12-22%的CD4+细胞和5-13%的CD8+细胞表达IL-2mRNA，而这两个亚群中30-50%的细胞表达IL-2-RmRNA。IL-2颗粒计数与mRNA/细胞水平有关，在有贴壁辅助细胞存在的情况下，CD4+亚群的IL-2颗粒数较高，而CD8+亚群的IL-2颗粒数可增加数倍。使用PMA作为辅助刺激，除贴壁细胞外，还能显著增加IL-2mRNA阳性淋巴细胞的频率和培养上清液中IL-2的活性，但对增殖反应无明显促进作用。这些结果表明，在单细胞水平上，许多CD_4~+或CD_8~+淋巴细胞可以产生IL-2 m RNA，CsA可抑制IL-2的产生，而PMA则能增强IL-2的诱导作用。
RNA-RNA in situ hybridization was used to study the frequency of cells producing mRNA for IL-2 and IL-2-R in T lymphocytes stimulated by either of three mitogens: anti-CD3, anti-CD28, or PHA. Both CD4+ and CD8+ T cells expressed transcripts for IL-2 and the low-affinity IL-2-R when stimulated with these mitogens plus PMA. IL-2 transcripts peaked at 8-16 h, and IL-2-R at 24-40 h. Cyclosporin A (CSA) inhibited the synthesis of IL-2, but not IL-2-R mRNA, after stimulation by PHA or anti-CD3. However, higher concentrations of CSA were necessary to achieve 50-70% inhibition after stimulation with anti-CD28. At optimal points 12-22% of CD4+ and 5-13% of CD8+ cells expressed IL-2 mRNA, while 30-50% of cells of both subsets had IL-2-R mRNA. The IL-2 grain counts, which relate to the level of mRNA/cell, were higher in the CD4+ subset but could be increased several fold in the CD8+ subset in the presence of adherent accessory cells. The use of PMA as an accessory stimulus, in addition to adherent cells, greatly increased the frequency of lymphocytes with IL-2 mRNA and the amount of IL-2 activity in the culture medium, but the proliferative response was not significantly boosted. These observations indicate at the single cell level that many CD4+ or CD8+ lymphocytes can make IL-2 mRNA, and that the induction of IL-2 with several stimuli is reduced by CSA and enhanced by PMA.