Use of the human monocytic leukemia THP-1 cell line and co-incubation with microsomes to identify and differentiate hapten and prohapten sensitizers
Use of the human monocytic leukemia THP-1 cell line and co-incubation with microsomes to identify and differentiate hapten and prohapten sensitizers
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DOI：
10.1016/j.tox.2010.12.004
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发表时间：
2011-02-27
期刊：
影响因子：
4.5
通讯作者：
Siegel, Paul D.
中科院分区：
文献类型：
--
作者：
Chipinda, Itai;Ruwona, Tinashe B.;Siegel, Paul D.
消费品和医疗产品可能含有可沥滤的化学过敏原，可导致皮肤过敏。最近的努力已经针对开发非基于动物的测试，例如用于鉴定皮肤致敏物的体外细胞活化测定。由于缺乏前半抗原的生物活化，通过体外测定的前半抗原鉴定仍然存在问题。本研究评价了半抗原和前半抗原暴露对人单核细胞白血病THP-1细胞系中细胞表面标志物（CD 86、CD 54和CD 40）表达的影响。激活和共刺激标志物的上调是过敏性致敏过程中的关键事件，并已被报道用作皮肤致敏的指标。将细胞暴露于浓度范围为1至10 μ M的前半抗原苯并（a）芘（BaP）、7，12-二甲基苯并（a）蒽（DMBA）、香芹酮肟（考克斯）、肉桂醇（CA）和异丁香酚（IEG）24和48小时。使用直接结合半抗原二硝基氯苯（DNCB）、苯醌（BQ）、丙烯酸羟乙酯（HEA）和溴化苄（BB）作为阳性对照。细胞也暴露于刺激物十二烷基硫酸钠（SOS）和磺胺（SFA）。通过向细胞培养物中加入阿罗氯诱导的大鼠肝微粒体（S9）实现前半抗原的生物活化。在用直接作用半抗原（+/- S9）或前半抗原（+S9）处理的THP-1细胞中观察到CD 86、CD 54（ICAM-1）和CD 40的表面表达的一致上调。在不存在外源性添加S9的情况下，暴露于皮肤刺激物或前半抗原后未观察到这些标志物的上调。总之，修改体外细胞培养试验，包括与微粒体共孵育，增强了前半抗原的鉴定，并使其能够与直接结合的半抗原明确区分。出版社：Elsevier爱尔兰Ltd.
Consumer and medical products can contain leachable chemical allergens which can cause skin sensitization. Recent efforts have been directed at the development of non-animal based tests such as in vitro cell activation assays for the identification of skin sensitizers. Prohapten identification by in vitro assays is still problematic due to the lack of prohapten bioactivation. The present study evaluated the effect of hapten and prohapten exposure on cell surface markers expression (CD86, CD54 and CD40) in the human monocytic leukemia, THP-1, cell line. Upregulation of activation and costimulatory markers are key events in the allergic sensitization process and have been reported to serve as indicators of skin sensitization. Cells were exposed to the prohaptens benzo(a)pyrene (BaP), 7,12-dimethylbenz(a)anthracene (DMBA), carvone oxime (COx), cinnamic alcohol (CA) and isoeugenol (IEG) at concentrations ranging from 1 to 10 mu M for 24 and 48 h. The direct-binding haptens dinitrochlorobenzene (DNCB), benzoquinone (BQ), hydroxylethyl acrylate (HEA) and benzylbromide (BB) were used as positive controls. Cells were also exposed to the irritants sodium dodecyl sulfate (SOS) and sulfanilamide (SFA). Bioactivation of prohaptens was achieved by adding aroclor-induced rat liver microsomes (S9) to the cell cultures. Consistent upregulation of surface expressions of CD86, CD54 (ICAM-1) and CD40 was observed in THP-1 cells treated with direct-acting haptens (+/- S9) or prohapten (+S9). Upregulation of these markers was not observed after exposure to skin irritants or prohaptens in the absence of exogenously added S9. In conclusion, modification of in vitro cell culture assays to include co-incubation with microsomes enhances identification of prohaptens and allows them to be clearly distinguished from direct-binding haptens. Published by Elsevier Ireland Ltd.