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Effect of IFN-γ on the kynurenine/tryptophan ratio in monolayer-cultured keratinocytes and a 3D reconstructed human epidermis model
Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.ORCID iD: 0000-0001-9847-5132
Malmö University, Faculty of Odontology (OD).
Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV). Malmö University, Biofilms Research Center for Biointerfaces.
Malmö University, Biofilms Research Center for Biointerfaces. Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).ORCID iD: 0000-0003-0304-7528
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2020 (English)In: Journal of dermatological science (Amsterdam), ISSN 0923-1811, E-ISSN 1873-569X, Vol. 99, no 3, p. 177-184, article id S0923-1811(20)30234-6Article in journal (Refereed) Published
Abstract [en]

BACKGROUND: Interferon-gamma (IFN-γ) represents a potent inducer for keratinocyte inflammatory and immune activation in vitro. Since tryptophan (trp) conversion to kynurenine (kyn) is involved in inflammation, the topical kyn/trp ratio may serve as a biomarker of skin inflammation. However, the trp metabolism in keratinocytes exposed to IFN-γ is not yet fully understood.

OBJECTIVE: The aim of this study was to establish a human epidermis model in order to quantify cytokine and kyn/trp secretion from IFN-γ stimulated cells and tissues. Moreover, to compare the cell response of 2D-cultured keratinocytes and the 3D epidermis model.

METHODS: Polycarbonate filters were used on which primary keratinocytes could attach and stratify in order to form the typical layers of reconstructed human epidermis (RHE). After IFN-γ treatment, secretion of kyn/trp was measured by high performance liquid chromatography. Gene and protein expression of indoleamine 2,3-dioxygenase 1 (IDO) was analyzed with real-time PCR and immunohistochemistry. The secretion of cytokines was quantified with ELISA.

RESULTS: Trp catabolism to kyn was significantly increased (P < 0.01) in the 2D culture in response to IFN-γ treatment. Before kyn secretion, IDO was strongly upregulated (P < 0.001). IFN-γ treatment also increased the secretion of IL-6 and IL-8 from the keratinocytes. In the RHE, IDO was upregulated by IFN-γ, and kyn secretion could be detected. Interestingly, IDO expression was only present in the basal cells of the RHE.

CONCLUSION: Our results suggest that IFN-γ acts as an inducer of trp degradation preferentially in undifferentiated keratinocytes, indicated by the IDO expression in the basal layer of the RHE.

Place, publisher, year, edition, pages
Japanese Society for Investigative Dermatology , 2020. Vol. 99, no 3, p. 177-184, article id S0923-1811(20)30234-6
Keywords [en]
IDO, Kynurenine, Pro-inflammatory cytokines, Reconstructed human epidermis, Tryptophan
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:mau:diva-18014DOI: 10.1016/j.jdermsci.2020.07.005ISI: 000582365800005PubMedID: 32782183Scopus ID: 2-s2.0-85089257839OAI: oai:DiVA.org:mau-18014DiVA, id: diva2:1458881
Available from: 2020-08-18 Created: 2020-08-18 Last updated: 2024-02-05Bibliographically approved
In thesis
1. Non-invasive monitoring of low molecular weight biomarkers relevant to skin inflammation and cancer
Open this publication in new window or tab >>Non-invasive monitoring of low molecular weight biomarkers relevant to skin inflammation and cancer
2021 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Development of skin inflammation and cancer in viable epidermis and dermis involve slow molecular weight (LMW) metabolites. We hypothesize that these LMW compounds can be collected on the surface of the skin and used for non-invasive diagnostics of skin disorders. Keeping in mind that substantial transdermal penetrationis achieved only for molecules of < 500 Da, we focused on topical monitoring of LMW biomarkers. In this thesis we investigated non-invasive, topical methods for monitoring LMW biomarkers by relevant in vitro and in vivo experiments. The LMW biomarkers were:

- reactive oxygen species (ROS), specifically, hydrogen peroxide, H2O2

- amino acids and their derivatives, i.e., tryptophan (Trp), kynurenine (Kyn; a Trpderivative), phenylalanine (Phe), and tyrosine (Tyr; a Phe derivative).

Initially, we have carried out in vitro experiments using dermatomed porcine skin and cell cultures. We characterized permeability of the biomarkers through skin and assessed methods of their monitoring. By using Prussian white particles, deposited on porcine skin, we demonstrated that hydrophilic biomarkers, such as H2O2, permeate the skin mainly through hair follicle pathways (Paper I). In paper II, we have showed that the enzymes transforming Trp to the inflammation and cancer biomarker Kyn, are expressed in the basal layer of epidermis. The magnitude of changes of the Trp/Kyn ratio in the cell culture model was assessed. In paper III, we have characterized Trp and Kyn permeability through skin in vitro, concluding that their permeabilities through stratum corneum are comparable. By in vivo experiments outlined in Paper IV, we have demonstrated the feasibility of topical, non-invasive sampling of Trp and Kyn, in relation to other amino acids. Kyn detection was compromised by its low abundance on the skin. In paper V, we performed a proof-of-concept study in vivo and confirmed that non-invasive sampling of Trp and amino acids of similar abundance, such as Phe and Tyr, is more robust. We concluded that Phe/Trp ratio might be equally good biomarker of skin disorders as a predicted Trp/Kyn ratio. Summarizing, the results of this thesis provide basic knowledge for deeper clinical studies of non-invasive, topical sampling of hydrophilic LMW biomarkers of skin inflammation and cancer.

Place, publisher, year, edition, pages
Malmö: Malmö universitet, 2021. p. 66
Series
Malmö University Health and Society Dissertations, ISSN 1653-5383 ; 2021:9
Keywords
Tryptophan, Kynurenine, Phenylalanine, Tyrosine, Low molecular Weight Biomarkers, Non-Invasive, Skin Surface Sampling, Skin Permeability, Hydrogels, Prussian White
National Category
Health Sciences
Research subject
Health and society
Identifiers
urn:nbn:se:mau:diva-46859 (URN)10.24834/isbn.9789178772230 (DOI)978-91-7877-222-3 (ISBN)978-91-7877-223-0 (ISBN)
Public defence
2021-11-29, Live stream and on location at HS assembly hall, Jan Waldenströms gata 25, ö, 09:00
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Supervisors
Note

Paper III published in dissertation as manuscript with title Non-invasive, Topical Sampling of Potential Skin Cancer Biomarkers,Kynurenine and Tryptophan: Study on Healthy Volunteers

Available from: 2021-11-15 Created: 2021-11-15 Last updated: 2022-11-09Bibliographically approved

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Gustafsson, AnnaPrgomet, ZdenkaJankovskaja, SkaidreRuzgas, TautgirdasEngblom, JohanOhlsson, LarsGjörloff Wingren, Anette

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Gustafsson, AnnaPrgomet, ZdenkaJankovskaja, SkaidreRuzgas, TautgirdasEngblom, JohanOhlsson, LarsGjörloff Wingren, Anette
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