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Natural NADH and FAD Autofluorescence as Label-Free Biomarkers for Discriminating Subtypes and Functional States of Immune Cells

Immune cell activity is a major factor for disease progression in inflammatory bowel diseases (IBD). Classifying the type and functional state of immune cells is therefore crucial in clinical diagnostics of IBD. Label-free optical technologies exploiting NADH and FAD autofluorescence, such as multip...

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Autores principales: Lemire, Sarah, Thoma, Oana-Maria, Kreiss, Lucas, Völkl, Simon, Friedrich, Oliver, Neurath, Markus F., Schürmann, Sebastian, Waldner, Maximilian J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8880312/
https://www.ncbi.nlm.nih.gov/pubmed/35216453
http://dx.doi.org/10.3390/ijms23042338
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author Lemire, Sarah
Thoma, Oana-Maria
Kreiss, Lucas
Völkl, Simon
Friedrich, Oliver
Neurath, Markus F.
Schürmann, Sebastian
Waldner, Maximilian J.
author_facet Lemire, Sarah
Thoma, Oana-Maria
Kreiss, Lucas
Völkl, Simon
Friedrich, Oliver
Neurath, Markus F.
Schürmann, Sebastian
Waldner, Maximilian J.
author_sort Lemire, Sarah
collection PubMed
description Immune cell activity is a major factor for disease progression in inflammatory bowel diseases (IBD). Classifying the type and functional state of immune cells is therefore crucial in clinical diagnostics of IBD. Label-free optical technologies exploiting NADH and FAD autofluorescence, such as multiphoton microscopy, have been used to describe tissue morphology in healthy and inflamed colon samples. Nevertheless, a strategy for the identification of single immune cell subtypes within the tissue is yet to be developed. This work aims to initiate an understanding of autofluorescence changes depending on immune cell type and activation state. For this, NADH and FAD autofluorescence signals of different murine immune cell subtypes under native conditions, as well as upon in vitro stimulation and cell death, have been evaluated. Autofluorescence was assessed using flow cytometry and multiphoton microscopy. Our results reveal significantly increased NADH and FAD signals in innate immune cells compared to adaptive immune cells. This allowed identification of relative amounts of neutrophils and CD4+ T cells in mixed cell suspensions, by using NADH signals as a differentiation marker. Furthermore, in vitro stimulation significantly increased NADH and FAD autofluorescence in adaptive immune cells and macrophages. Cell death induced a significant drop in NADH autofluorescence, while FAD signals were hardly affected. Taken together, these results demonstrate the value of autofluorescence as a tool to characterize immune cells in different functional states, paving the way to the label-free clinical classification of IBD in the future.
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spelling pubmed-88803122022-02-26 Natural NADH and FAD Autofluorescence as Label-Free Biomarkers for Discriminating Subtypes and Functional States of Immune Cells Lemire, Sarah Thoma, Oana-Maria Kreiss, Lucas Völkl, Simon Friedrich, Oliver Neurath, Markus F. Schürmann, Sebastian Waldner, Maximilian J. Int J Mol Sci Article Immune cell activity is a major factor for disease progression in inflammatory bowel diseases (IBD). Classifying the type and functional state of immune cells is therefore crucial in clinical diagnostics of IBD. Label-free optical technologies exploiting NADH and FAD autofluorescence, such as multiphoton microscopy, have been used to describe tissue morphology in healthy and inflamed colon samples. Nevertheless, a strategy for the identification of single immune cell subtypes within the tissue is yet to be developed. This work aims to initiate an understanding of autofluorescence changes depending on immune cell type and activation state. For this, NADH and FAD autofluorescence signals of different murine immune cell subtypes under native conditions, as well as upon in vitro stimulation and cell death, have been evaluated. Autofluorescence was assessed using flow cytometry and multiphoton microscopy. Our results reveal significantly increased NADH and FAD signals in innate immune cells compared to adaptive immune cells. This allowed identification of relative amounts of neutrophils and CD4+ T cells in mixed cell suspensions, by using NADH signals as a differentiation marker. Furthermore, in vitro stimulation significantly increased NADH and FAD autofluorescence in adaptive immune cells and macrophages. Cell death induced a significant drop in NADH autofluorescence, while FAD signals were hardly affected. Taken together, these results demonstrate the value of autofluorescence as a tool to characterize immune cells in different functional states, paving the way to the label-free clinical classification of IBD in the future. MDPI 2022-02-20 /pmc/articles/PMC8880312/ /pubmed/35216453 http://dx.doi.org/10.3390/ijms23042338 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lemire, Sarah
Thoma, Oana-Maria
Kreiss, Lucas
Völkl, Simon
Friedrich, Oliver
Neurath, Markus F.
Schürmann, Sebastian
Waldner, Maximilian J.
Natural NADH and FAD Autofluorescence as Label-Free Biomarkers for Discriminating Subtypes and Functional States of Immune Cells
title Natural NADH and FAD Autofluorescence as Label-Free Biomarkers for Discriminating Subtypes and Functional States of Immune Cells
title_full Natural NADH and FAD Autofluorescence as Label-Free Biomarkers for Discriminating Subtypes and Functional States of Immune Cells
title_fullStr Natural NADH and FAD Autofluorescence as Label-Free Biomarkers for Discriminating Subtypes and Functional States of Immune Cells
title_full_unstemmed Natural NADH and FAD Autofluorescence as Label-Free Biomarkers for Discriminating Subtypes and Functional States of Immune Cells
title_short Natural NADH and FAD Autofluorescence as Label-Free Biomarkers for Discriminating Subtypes and Functional States of Immune Cells
title_sort natural nadh and fad autofluorescence as label-free biomarkers for discriminating subtypes and functional states of immune cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8880312/
https://www.ncbi.nlm.nih.gov/pubmed/35216453
http://dx.doi.org/10.3390/ijms23042338
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