In vitro detection of cytotoxic T and NK cells in peripheral blood of patients with various drug-induced skin diseases
In vitro detection of cytotoxic T and NK cells in peripheral blood of patients with various drug-induced skin diseases
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DOI：
10.1111/j.1398-9995.2009.02180.x
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发表时间：
2010-03-01
期刊：
影响因子：
12.4
通讯作者：
Pichler, W. J.
中科院分区：
文献类型：
--
作者：
Zawodniak, A.;Lochmatter, P.;Pichler, W. J.
P>背景：细胞毒性细胞参与大多数形式的药物引起的皮肤病。到目前为止，还没有体外测试解决药物过敏反应的这一方面。我们的报告评估了是否可以在不同形式药物过敏的非急性患者的外周血中检测到药物诱导的细胞毒性细胞，以及体外检测这些细胞是否有助于药物过敏的诊断。 方法：对 12 例药物过敏患者和 16 例药物过敏患者的外周血单核细胞进行颗粒酶 B 酶联免疫斑点形成（ELISPOT）和细胞表面脱颗粒标志物 CD107a 的表达评估。 drug-exposed healthy controls.Results:In 10/12 allergic patients culprit but not irrelevant drug elicited granzymeB release after 48-72 h stimulation.在淋巴细胞转化试验中测量，在对该药物具有高增殖反应的患者中，其结果明显呈阳性。在颗粒酶 B ELISPOT 中表现出中度或低度增殖和低药物反应的患者中，与白细胞介素 (IL)-7/IL-15 过夜预孵育可增强药物特异性颗粒酶 B 释放，并允许清楚地识别出违规物质。 CD4+/CD3+、CD8+/CD3+ T 细胞以及 CD56+/CD3- 自然杀伤细胞的 CD107a 染色呈阳性。接触药物的健康捐献者均未对测试药物产生反应，而过敏患者仅对违规药物产生反应，而对耐受药物没有反应。结论：GranzymeB ELISPOT 是一种高度特异性的体外方法，可检测药物过敏患者外周血中药物反应的细胞毒性细胞，甚至在疾病出现数年后也可检测。与 IL-7/IL-15 预孵育一起，即使对于某些增殖药物反应较弱的患者，也可能有助于识别违规药物。
P>Background:Cytotoxic cells are involved in most forms of drug-induced skin diseases. Till now, no in vitro test addressed this aspect of drug-allergic responses. Our report evaluates whether drug-induced cytotoxic cells can be detected in peripheral blood of nonacute patients with different forms of drug hypersensitivity, and also whether in vitro detection of these cells could be helpful in drug-allergy diagnosis.Methods:GranzymeB enzyme-linked immunosorbent spot-forming (ELISPOT) and cell surface expression of the degranulation marker CD107a were evaluated on peripheral blood mononuclear cells from 12 drug-allergic patients in remission state and 16 drug-exposed healthy controls.Results:In 10/12 allergic patients culprit but not irrelevant drug elicited granzymeB release after 48-72 h stimulation. It was clearly positive in patients with high proliferative response to the drug, measured in lymphocyte transformation tests. In patients, who showed moderate or low proliferation and low drug-response in granzymeB ELISPOT, overnight preincubation with interleukin (IL)-7/IL-15 enhanced drug-specific granzymeB release and allowed to clearly identify the offending agent. CD107a staining was positive on CD4+/CD3+, CD8+/CD3+ T cells as well as CD56+/CD3- natural killer cells. None of the drug-exposed healthy donors reacted to the tested drugs and allergic patients reacted only to the offending, but not to tolerated drugs.Conclusion:GranzymeB ELISPOT is a highly specific in vitro method to detect drug-reacting cytotoxic cells in peripheral blood of drug-allergic patients even several years after disease manifestation. Together with IL-7/IL-15 preincubation, it may be helpful in indentifying the offending drug even in some patients with weak proliferative drug-response.