Modification of cell-surface thiols elicits activation of human monocytic cell line THP-1: Possible involvement in effect of haptens 2,4-dinitrochlorobenzene and nickel sulfate
Modification of cell-surface thiols elicits activation of human monocytic cell line THP-1: Possible involvement in effect of haptens 2,4-dinitrochlorobenzene and nickel sulfate
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DOI：
10.2131/jts.34.139
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发表时间：
2009-04-01
影响因子：
2
通讯作者：
Itagaki, Hiroshi
中科院分区：
文献类型：
--
作者：
Hirota, Morihiko;Suzuki, Mie;Itagaki, Hiroshi
人单核细胞系THP-1细胞作为体外皮肤致敏试验的指标。尽管p38丝裂原活化蛋白激酶（MAPKs）和细胞内氧化还原失衡在皮肤致敏剂激活THP-1中起着至关重要的作用，但细胞激活的触发因素尚未确定。因此，我们研究了半抗原是否直接或间接诱导THP-1成熟。2,4-二硝基氯苯（DNCB）诱导CD86表达，而非二硝基酚（DNP）偶联的牛血清白蛋白或DNP偶联的胎牛血清。DNCB和硫酸镍（NiSO4）也诱导了细胞表面硫醇和p38 MAPK磷酸化的相关变化。然而，DNCB具有膜渗透性，因此其直接作用可能不局限于细胞膜蛋白。接下来，我们发现CD86的表达和巨噬细胞炎症蛋白-1 β (MIP-1 β)的产生被膜不渗透硫醇阻断剂5,5'-二硫代obis（2-硝基苯甲酸）（DTNB）增强，这些变化被p38 MAPK途径的抑制剂SB203580抑制。最后，我们证实了牛血清白蛋白（BSA） Alexa Fluor 488偶联物的内吞活性不影响THP-1细胞表面的硫醇。因此，我们的数据表明，细胞表面硫醇的变化是成熟的触发因素之一，并在半抗原激活THP-1细胞中发挥关键作用。
Human monocytic cell line THP-1 cells are used as an indicator for in vitro skin sensitization testing. Although p38 mitogen-activated protein kinases (MAPKs) and intracellular redox imbalance play crucial roles in the activation of THP-1 by skin sensitizers, the trigger of cell activation has not been identified. Therefore, we examined whether haptens induce THP-1 maturation directly or indirectly. 2,4-Dinitrochlorobenzene (DNCB), but not dinitrophenol (DNP)-conjugated bovine serum albumin or DNP-conjugated fetal bovine serum, induced CD86 expression. DNCB and nickel sulfate (NiSO4) also induced related changes of cell-surface thiols and phosphorylation of p38 MAPK. However, DNCB is membrane-permeable, and so its direct effect may not be confined to cell membrane proteins. Next, we found that CD86 expression and macrophage inflammatory protein-1 beta (MIP-1 beta) production were augmented by the membrane-impermeable thiol blocker 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), and these changes were suppressed by an inhibitor of the p38 MAPK pathway, SB203580. Finally, we confirmed that endocytotic activity for bovine serum albumin (BSA) Alexa Fluor 488 conjugate did not affect cell-surface thiols on THP-1 cells. Thus, our data indicate that the changes of cell-surface thiols are one of the triggers of maturation, and play a key role in activation of THP-1 cells by haptens.