Medical Department III, Hospital Großhadern, Ludwig-Maximilians-University, 81377 Munich, Germany.
Department of Pulmonary Rehabilitation, German Center for Lung Research (DZL), Phillipps-University of Marburg, 35043 Marburg, Germany.
Biomolecules. 2023 Jun 14;13(6):989. doi: 10.3390/biom13060989.
Volatile organic compounds (VOCs) reflect the metabolism in healthy and pathological conditions, and can be collected easily in a noninvasive manner. They are directly measured using electronical nose (eNose), and may qualify as a systemic tool to monitor biomarkers related to disease. Myeloid leukemic blasts can be transformed into leukemia-derived dendritic cells (DC) able to improve (anti-leukemic) immune responses. To profile immunological changes in healthy and acute myeloid leukemic (AML) patients' ex vivo cell cultures, we correlated the cell biological data with the profiles of cell culture supernatant-derived VOCs. DC/DC from leukemic or healthy whole blood (WB) were generated without (Control) or with immunomodulatory Kit M (Granulocyte macrophage-colony-stimulating-factor (GM-CSF) + prostaglandin E (PGE)) in dendritic cell cultures (DC culture). Kit-pretreated/not pretreated WB was used to stimulate T cell-enriched immunoreactive cells in mixed lymphocyte cultures (MLC culture). Leukemia-specific adaptive and innate immune cells were detected with a degranulation assay (Deg) and an intracellular cytokine assay (InCyt). Anti-leukemic cytotoxicity was explored with a cytotoxicity fluorolysis assay (CTX). VOCs collected from serum or DC- and MLC culture supernatants (with vs. without Kit M pretreatment and before vs. after culture) were measured using eNose. Compared to the Control (without treatment), Kit M-pretreated leukemic and healthy WB gave rise to higher frequencies of mature (leukemia-derived) DC subtypes of activated and (memory) T cells after MLC. Moreover, antigen (leukemia)-specific cells of several lines (innate and adaptive immunity cells) were induced, giving rise to blast-lysing cells. The eNose could significantly distinguish between healthy and leukemic patients' serum, DC and MLC culture supernatant-derived volatile phases and could significantly separate several supernatant (with vs. without Kit M treatment, cultured vs. uncultured)-derived VOCs within subgroups (healthy DC or leukemic DC, or healthy MLC or leukemic MLC supernatants). Interestingly, the eNose could indicate a Kit M- and culture-associated effect. The eNose may be a prospective option for the deduction of a VOC-based profiling strategy using serum or cell culture supernatants and could be a useful diagnostic tool to recognize or qualify AML disease.
挥发性有机化合物（VOCs）反映了健康和病理条件下的代谢情况，并且可以以非侵入性的方式轻松收集。它们使用电子鼻（eNose）直接测量，可以作为监测与疾病相关生物标志物的系统工具。髓样白血病blasts 可以转化为具有改善（抗白血病）免疫反应能力的白血病衍生树突状细胞（DC）。为了分析健康和急性髓细胞性白血病（AML）患者体外细胞培养物中的免疫学变化，我们将细胞生物学数据与细胞培养上清液衍生 VOCs 的图谱相关联。从白血病或健康全血（WB）生成无（对照）或有免疫调节 Kit M（粒细胞巨噬细胞集落刺激因子（GM-CSF）+前列腺素 E（PGE））的 DC/DC 在树突状细胞培养物（DC 培养物）中。用 Kit 预处理/未预处理的 WB 刺激混合淋巴细胞培养物（MLC 培养物）中的 T 细胞富集免疫反应性细胞。使用脱颗粒测定（Deg）和细胞内细胞因子测定（InCyt）检测白血病特异性适应性和先天免疫细胞。使用细胞毒性荧光溶解测定（CTX）探索抗白血病细胞毒性。使用 eNose 测量来自血清或 DC 和 MLC 培养物上清液（有 vs. 无 Kit M 预处理以及培养前后）的 VOCs。与对照（无处理）相比，Kit M 预处理的白血病和健康 WB 在 MLC 后产生更高频率的成熟（白血病衍生）DC 亚型和激活的（记忆）T 细胞。此外，诱导了几种（先天和适应性免疫细胞）抗原（白血病）特异性细胞，导致blasts 裂解细胞。eNose 可以显著区分健康和白血病患者的血清、DC 和 MLC 培养上清液衍生的挥发性相，并可以在亚组（健康 DC 或白血病 DC，或健康 MLC 或白血病 MLC 上清液）内显著分离几个上清液（有 vs. 无 Kit M 处理、培养 vs. 未培养）衍生的 VOCs。有趣的是，eNose 可以指示 Kit M 和培养相关的影响。eNose 可能是使用血清或细胞培养上清液进行基于 VOC 的分析策略的有前途的选择，并且可以成为识别或定性 AML 疾病的有用诊断工具。
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