Andreas Glässner, Michael Steffens, Amol Fatangare, Gerda Wurpts, Per Hoffmann, Philipp N Deck, Christine Krämer, Stefani Röseler, Albert Sickmann, Markus M Nöthen, Amir S Yazdi, Bernhardt Sachs
{"title":"用于体外检测药物致敏性的 PBMC 差异基因表达分析。","authors":"Andreas Glässner, Michael Steffens, Amol Fatangare, Gerda Wurpts, Per Hoffmann, Philipp N Deck, Christine Krämer, Stefani Röseler, Albert Sickmann, Markus M Nöthen, Amir S Yazdi, Bernhardt Sachs","doi":"10.1016/j.alit.2024.11.006","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusions: </strong>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.</p>","PeriodicalId":48861,"journal":{"name":"Allergology International","volume":" ","pages":""},"PeriodicalIF":6.2000,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Analysis of differential gene expression of PBMC for the in vitro detection of drug sensitization.\",\"authors\":\"Andreas Glässner, Michael Steffens, Amol Fatangare, Gerda Wurpts, Per Hoffmann, Philipp N Deck, Christine Krämer, Stefani Röseler, Albert Sickmann, Markus M Nöthen, Amir S Yazdi, Bernhardt Sachs\",\"doi\":\"10.1016/j.alit.2024.11.006\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>The detection of drug-specific activation of T cells in the lymphocyte transformation test (LTT) is mainly based on cell proliferation or cytokine secretion. 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Analysis of differential gene expression of PBMC for the in vitro detection of drug sensitization.
Background: 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.
Methods: 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.
Results: 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.
Conclusions: 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.
期刊介绍:
Allergology International is the official journal of the Japanese Society of Allergology and publishes original papers dealing with the etiology, diagnosis and treatment of allergic and related diseases. Papers may include the study of methods of controlling allergic reactions, human and animal models of hypersensitivity and other aspects of basic and applied clinical allergy in its broadest sense.
The Journal aims to encourage the international exchange of results and encourages authors from all countries to submit papers in the following three categories: Original Articles, Review Articles, and Letters to the Editor.