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Volatile Phases Derived from Serum, DC, or MLC Culture Supernatants to Deduce a VOC-Based Diagnostic Profiling Strategy for Leukemic Diseases

Affiliation
Medical Department III, Hospital Großhadern, Ludwig-Maximilians-University, 81377 Munich, Germany
Baudrexler, Tobias;
Affiliation
Department of Pulmonary Rehabilitation, German Center for Lung Research (DZL), Phillipps-University of Marburg, 35043 Marburg, Germany
Boeselt, Tobias;
ORCID
0000-0003-2557-6404
Affiliation
Medical Department III, Hospital Großhadern, Ludwig-Maximilians-University, 81377 Munich, Germany
Li, Lin;
Affiliation
Medical Department III, Hospital Großhadern, Ludwig-Maximilians-University, 81377 Munich, Germany
Bohlscheid, Sophia;
Affiliation
Department of Pulmonary Rehabilitation, German Center for Lung Research (DZL), Phillipps-University of Marburg, 35043 Marburg, Germany
Boas, Ursel;
Affiliation
Department of Hematology and Oncology, University Hospital of Augsburg, 86156 Augsburg, Germany
Schmid, Christoph;
ORCID
0000-0002-5636-0043
Affiliation
Department of Hematology and Oncology, University Hospital of Augsburg, 86156 Augsburg, Germany
Rank, Andreas;
Affiliation
Department of Hematology and Oncology, Diaconia Hospital Stuttgart, 70176 Stuttgart, Germany
Schmohl, Jörg;
Affiliation
Department of Pulmonary Rehabilitation, German Center for Lung Research (DZL), Phillipps-University of Marburg, 35043 Marburg, Germany
Koczulla, Rembert;
Affiliation
Medical Department III, Hospital Großhadern, Ludwig-Maximilians-University, 81377 Munich, Germany
Schmetzer, Helga Maria

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 leu ) 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 leu 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 1 (PGE 1 )) 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.

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