Li M D, McAllen K, Sharp B M
Department of Pharmacology, University of Tennessee College of Medicine, Memphis, USA.
J Leukoc Biol. 1999 May;65(5):707-14. doi: 10.1002/jlb.65.5.707.
Previous studies have shown that low levels of delta opioid receptor (DOR) mRNA were detectable by reverse transcription polymerase chain reaction (RT-PCR) in unstimulated splenocytes from BALB/c female mice. This study demonstrates that DOR transcripts can be detected in freshly obtained splenocytes froin CD 1 female mice as well. The results of studies using quantitative competitive RT-PCR showed that DOR transcripts in splenic T cells increased from < 1 copy/cell to 22 and 42 copies/cell, respectively, after stimulation with anti-CD3-epsilon for 24 and 48 h compared to the level in freshly obtained T cells. In the presence of actinomycin D, anti-CD3-epsilon did not affect the rate of degradation of DOR mRNA, suggesting that its stability is not altered by anti-CD3-epsilon. After incubation with phorbol myristate acetate (PMA) and ionomycin, the expression of DOR mRNA in splenocytes and T cells was significantly reduced compared with unstimulated cells in culture. In addition, the inhibitory effect of PMA prevented anti-CD3-epsilon-stimulated DOR expression. These data suggest that signaling through the T cell receptor complex by anti-CD3-epsilon regulates DOR expression differently than PMA and ionomycin.
先前的研究表明,通过逆转录聚合酶链反应(RT-PCR)可在未受刺激的BALB/c雌性小鼠脾细胞中检测到低水平的δ阿片受体(DOR)mRNA。本研究表明,在新获得的CD1雌性小鼠脾细胞中也可检测到DOR转录本。使用定量竞争性RT-PCR的研究结果显示,与新获得的T细胞相比,用抗CD3-ε刺激24小时和48小时后,脾T细胞中的DOR转录本分别从<1拷贝/细胞增加到22拷贝/细胞和42拷贝/细胞。在放线菌素D存在的情况下,抗CD3-ε不影响DOR mRNA的降解速率,表明其稳定性不受抗CD3-ε的影响。在用佛波酯(PMA)和离子霉素孵育后,与培养中未受刺激的细胞相比,脾细胞和T细胞中DOR mRNA的表达显著降低。此外,PMA的抑制作用阻止了抗CD3-ε刺激的DOR表达。这些数据表明,抗CD3-ε通过T细胞受体复合物发出的信号对DOR表达的调节与PMA和离子霉素不同。