Glässner Andreas, Steffens Michael, Fatangare Amol, Wurpts Gerda, Hoffmann Per, Deck Philipp N, Krämer Christine, Röseler Stefani, Sickmann Albert, Nöthen Markus M, Yazdi Amir S, Sachs Bernhardt
Research Division, Federal Institute for Drugs and Medical Devices (BfArM), Bonn, Germany.
Research Division, Federal Institute for Drugs and Medical Devices (BfArM), Bonn, Germany.
Allergol Int. 2025 Jul;74(3):414-423. doi: 10.1016/j.alit.2024.11.006. Epub 2025 Jan 11.
The detection of drug-specific activation of T cells in the lymphocyte transformation test (LTT) is mainly based on cell proliferation or cytokine secretion. However, the LTT presents with a varying sensitivity and specificity. The aim of our study was to analyse the genome wide gene expression of PBMC to identify drug allergy-specific gene regulation patterns. Additionally, gene expression differences related to the allergy-inducing drug or the type of allergy (immediate/delayed) were investigated.
Blood samples from 40 patients with allergy to different drugs and from 40 non-drug-allergic controls were recruited. PBMC were isolated and stimulated with the culprit drug ("LTT-platform"). Transcriptome analyses of PBMC were conducted after 3.5 days. The concentration of IFN-γ and IL-5 in the culture supernatants was measured by ELISA after 6 days.
The transcriptome analyses revealed an allergy type and drug-specific gene regulation in PBMC from patients. Importantly, in the corresponding control groups these genes were barely or even opposingly regulated. It was also shown that in particular cefuroxime exerted a strong effect on the gene regulation in PBMC. Quantitative RT-PCR of selected genes identified by the transcriptome analyses revealed that CCL17, CISH and CD25 were specifically upregulated in patients. Notably, a strong upregulation of CCL17, CISH or CD25 did not necessarily correlate with the ELISA outcome of the patients and controls.
Our results could be helpful for the identification of one or a panel of regulated genes considering patient specific parameters like the type of the allergy-inducing drug.
淋巴细胞转化试验（LTT）中T细胞药物特异性激活的检测主要基于细胞增殖或细胞因子分泌。然而，LTT的敏感性和特异性各不相同。我们研究的目的是分析外周血单核细胞（PBMC）的全基因组基因表达，以确定药物过敏特异性的基因调控模式。此外，还研究了与诱发过敏的药物或过敏类型（速发型/迟发型）相关的基因表达差异。
招募了40名对不同药物过敏的患者和40名非药物过敏对照的血样。分离PBMC并用致病药物进行刺激（“LTT平台”）。3.5天后对外周血单核细胞进行转录组分析。6天后通过酶联免疫吸附测定法（ELISA）测量培养上清液中γ干扰素（IFN-γ）和白细胞介素-5（IL-5）的浓度。
转录组分析揭示了患者外周血单核细胞中存在过敏类型和药物特异性的基因调控。重要的是，在相应的对照组中，这些基因几乎没有调控或甚至呈现相反的调控。还表明，特别是头孢呋辛对外周血单核细胞的基因调控有很强的作用。对转录组分析确定的选定基因进行定量逆转录聚合酶链反应（qRT-PCR）显示，CC趋化因子配体17（CCL17）、细胞因子诱导的含SH2结构域蛋白（CISH）和CD25在患者中特异性上调。值得注意的是，CCL17、CISH或CD25的强烈上调不一定与患者和对照的ELISA结果相关。
考虑到患者的特定参数，如诱发过敏的药物类型,我们的结果可能有助于识别一个或一组受调控的基因。
