Involvement of p-glycoprotein in the transmembrane transport of interleukin-2 (IL-2), IL-4, and interferon-gamma in normal human T lymphocytes
Involvement of p-glycoprotein in the transmembrane transport of interleukin-2 (IL-2), IL-4, and interferon-gamma in normal human T lymphocytes
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DOI：
10.1182/blood.v88.5.1747.bloodjournal8851747
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发表时间：
1996-09-01
期刊：
影响因子：
20.3
通讯作者：
Huber, H
中科院分区：
文献类型：
--
作者：
Drach, J;Gsur, A;Huber, H
由正常人T淋巴细胞表达的多药耐药P-糖蛋白（P-gp）的生理作用在很大程度上仍是未知的。为了研究P-gp是否参与细胞因子的转运，在不存在或存在P-gp抑制剂的情况下，用植物血凝素（PHA）刺激外周血淋巴细胞，并通过酶联免疫吸附试验定量这些培养物上清液中细胞因子（白细胞介素-2 [IL-2]、IL-4、IL-6、干扰素-γ [IFN-γ]）的浓度。P-gp抑制剂包括维拉帕米（Ver）、他莫昔芬（Tmx）和P-gp特异性单克隆抗体UIC 2。释放IL-2显着抑制这些抑制剂的浓度，也有效地阻止流出的罗丹明-123正常T淋巴细胞。淋巴细胞IL-2 mRNA表达在PHA对照组和P-gp抑制剂组之间无差异。Ver和Tmx不干扰T细胞活化，如通过CD 25和CD 69表达所确定的。在非血液学模型中，表达P-gp的HCT-8腺癌细胞系中，外源性添加IL-2显示对P-gp介导的罗丹明-123外排产生抑制作用。此外，检查了IL-2通过电生理紧密和极化HCT-8单层的跨上皮转运。观察到IL-2穿过致密单层的时间依赖性通量，其被Ver部分抑制。我们还研究了P-gp抑制剂是否抑制由活化的T细胞产生的其他细胞因子（IL-4、IL-6、IFN γ）的释放。IL-4和IFN-γ的释放被Ver、Tmx和UIC 2显著抑制;然而，IL-6的释放保持不受影响。这些数据显示了T淋巴细胞和HCT-8细胞中P-gp介导的IL-2跨膜通量。我们的结论是，P-gp参与正常外周血T淋巴细胞的细胞因子（IL-2，IL-4和IFN-γ）的运输。(C)1996年，美国血液学会。
The physiological role of the multidrug resistance P-glycoprotein (P-gp), which is expressed by normal human T lymphocytes, is still largely unknown. To investigate whether or not P-gp is involved in the transport of cytokines, peripheral blood lymphocytes were stimulated with phytohemagglutinin (PHA) in the absence or presence of P-gp inhibitors, and concentrations of cytokines (interleukin-2 [IL-2], IL-4, IL-6, interferon-gamma [IFN-gamma]) in the supernatants of these cultures were quantitated by enzyme-linked immunosorbent assay. P-gp inhibitors included verapamil (Ver), tamoxifen (Tmx), and the P-gp specific monoclonal antibody UIC2. Release of IL-2 was significantly suppressed by these inhibitors at concentrations that were also effective in blocking efflux of Rhodamine-123 from normal T lymphocytes. IL-2 mRNA expression in lymphocytes was not different between PHA control and the cultures with P-gp inhibitors. Ver and Tmx did not interfere with T-cell activation as determined by CD25 and CD69 expression. In a nonhematological model, the P-gp expressing HCT-8 adenocarcinoma cell line, exogenously added IL-2 was shown to exert an inhibitory effect on P-gp mediated Rhodamine-123 efflux. In addition, transepithelial transport of IL-2 by electrophysiologically tight and polarized HCT-8 monolayers was examined. A time-dependent flux of IL-2 across dense monolayers, which was partially inhibited by Ver, was observed. We also investigated whether or not P-gp inhibitors suppressed release of other cytokines produced by activated T cells (IL-4, IL-6, IFN gamma). Release of IL-4 and lFN-gamma was significantly inhibited by Ver, Tmx, and UIC2; however, release of IL-6 remained unaffected. These data show P-gp mediated transmembrane flux of IL-2 in T lymphocytes and HCT-8 cells. We conclude that P-gp participates in the transport of cytokines (IL-2, IL-4, and lFN-gamma) in normal peripheral T lymphocytes. (C) 1996 by The American Society of Hematology.