Universität zu Köln

Wolf, Julian, Zhuang, Xinyu, Hildebrand, Antonia, Boneva, Stefaniya, Schwaemmle, Melanie, Betancor, Paola Kammrath, Fan, Jiaqi, Boehringer, Daniel, Maier, Philip, Lange, Clemens, Reinhard, Thomas, Schlunck, Guenther and Lapp, Thabo ORCID: 0000-0001-7097-7587 (2020). Corneal tissue induces transcription of metallothioneins in monocyte-derived human macrophages. Mol. Immunol., 128. S. 188 - 195. OXFORD: PERGAMON-ELSEVIER SCIENCE LTD. ISSN 0161-5890

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Abstract

Purpose: Immune reactions following corneal transplantation are the most common cause of transplant failure. However, the underlying mechanisms of corneal graft rejection are not yet fully understood but increasing evidence points to a crucial role of the innate immune system in this context. Using a human in vitro model, we aimed to assess the response of human macrophages to stimulation with human corneal tissue and whether corneal endothelial cells (CEC) have immune-modulating properties. Methods: Human monocytes were isolated from peripheral blood mononuclear cells and differentiated into monocyte-derived macrophages (MDM). A standardized protocol was used for disaggregation of human corneas into fragments of defined sizes. MDMs were stimulated using processed corneal material with or without CEC. Lipopolysaccharide (LPS) or interferon-gamma (IFN gamma) served as controls. RNA sequencing was applied to analyze the impact of differential stimulation of MDMs on their transcriptional profile. RNA sequencing results were validated using digital PCR. Results: The transcriptional profile of MDMs was significantly modulated by the type of stimulus used for MDM activation as well as by the individual MDM donor. LPS- or IFN gamma-stimulation resulted in distinct transcriptional alterations compared to unstimulated MDMs including an upregulation of various cytokines such as CCL3, 4, 5, 19 or CXCL9. Corneal tissue induced the differential expression of 45 genes when compared to unstimulated MDMs, with several metallothioneins (MTs) among the upregulated factors (MT1A, MT1E, MT1F, MT1G, MT1H, MT1L, MT1M, MT1X, MT2A). This effect was independent of the presence or absence of CEC. PCR validation confirmed induction of 3 different metallothioneins (MT1G, MT1H and MT2A) in MDMs stimulated by corneal tissue. Conclusions: The MDM in vitro model proved to be a robust tool to study the effects of LPS, IFN. and corneal tissue homogenates on the transcriptional activity of MDM. Human macrophages showed a distinct upregulation of various MTs when challenged with human corneal allogen with or without corneal endothelium, which might have an immune-modulatory effect. As a general observation, it appears that in MDM-based studies a significant donor-dependent effect on the transcriptional profile of MDMs needs to be considered and adjusted before downstream analysis.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Wolf, Julian UNSPECIFIED UNSPECIFIED UNSPECIFIED Zhuang, Xinyu UNSPECIFIED UNSPECIFIED UNSPECIFIED Hildebrand, Antonia UNSPECIFIED UNSPECIFIED UNSPECIFIED Boneva, Stefaniya UNSPECIFIED UNSPECIFIED UNSPECIFIED Schwaemmle, Melanie UNSPECIFIED UNSPECIFIED UNSPECIFIED Betancor, Paola Kammrath UNSPECIFIED UNSPECIFIED UNSPECIFIED Fan, Jiaqi UNSPECIFIED UNSPECIFIED UNSPECIFIED Boehringer, Daniel UNSPECIFIED UNSPECIFIED UNSPECIFIED Maier, Philip UNSPECIFIED UNSPECIFIED UNSPECIFIED Lange, Clemens UNSPECIFIED UNSPECIFIED UNSPECIFIED Reinhard, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED Schlunck, Guenther UNSPECIFIED UNSPECIFIED UNSPECIFIED Lapp, Thabo UNSPECIFIED orcid.org/0000-0001-7097-7587 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-309463
DOI:	10.1016/j.molimm.2020.10.016
Journal or Publication Title:	Mol. Immunol.
Volume:	128
Page Range:	S. 188 - 195
Date:	2020
Publisher:	PERGAMON-ELSEVIER SCIENCE LTD
Place of Publication:	OXFORD
ISSN:	0161-5890
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language TRANSPLANTATION; SUPPRESSION; EXPRESSION Multiple languages Biochemistry & Molecular Biology; Immunology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/30946