A human 3D immune competent full-thickness skin model mimicking dermal dendritic cell activation

Johanna Maria Hölken; Katja Friedrich; Marion Merkel; Nelli Blasius; Ursula Engels; Timo Buhl; Karsten Rüdiger Mewes; Lars Vierkotten; Nicole Elisabeth Teusch

doi:10.3389/fimmu.2023.1276151

A human 3D immune competent full-thickness skin model mimicking dermal dendritic cell activation

Front Immunol. 2023 Nov 6:14:1276151. doi: 10.3389/fimmu.2023.1276151. eCollection 2023.

Authors

Johanna Maria Hölken¹, Katja Friedrich¹, Marion Merkel², Nelli Blasius², Ursula Engels², Timo Buhl³, Karsten Rüdiger Mewes², Lars Vierkotten², Nicole Elisabeth Teusch¹

Affiliations

¹ Institute of Pharmaceutical Biology and Biotechnology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany.
² Alternative Methods and Tissue Engineering, Henkel AG & Co. KGaA, Düsseldorf, Germany.
³ Department of Dermatology, Venereology and Allergology, University Medical Center Göttingen, Göttingen, Germany.

Abstract

We have integrated dermal dendritic cell surrogates originally generated from the cell line THP-1 as central mediators of the immune reaction in a human full-thickness skin model. Accordingly, sensitizer treatment of THP-1-derived CD14^-, CD11c⁺ immature dendritic cells (iDCs) resulted in the phosphorylation of p38 MAPK in the presence of 1-chloro-2,4-dinitrobenzene (DNCB) (2.6-fold) as well as in degradation of the inhibitor protein kappa B alpha (IκBα) upon incubation with NiSO₄ (1.6-fold). Furthermore, NiSO₄ led to an increase in mRNA levels of IL-6 (2.4-fold), TNF-α (2-fold) and of IL-8 (15-fold). These results were confirmed on the protein level, with even stronger effects on cytokine release in the presence of NiSO₄: Cytokine secretion was significantly increased for IL-8 (147-fold), IL-6 (11.8-fold) and IL-1β (28.8-fold). Notably, DNCB treatment revealed an increase for IL-8 (28.6-fold) and IL-1β (5.6-fold). Importantly, NiSO₄ treatment of isolated iDCs as well as of iDCs integrated as dermal dendritic cell surrogates into our full-thickness skin model (SM) induced the upregulation of the adhesion molecule clusters of differentiation (CD)54 (iDCs: 1.2-fold; SM: 1.3-fold) and the co-stimulatory molecule and DC maturation marker CD86 (iDCs ~1.4-fold; SM:~1.5-fold) surface marker expression. Noteworthy, the expression of CD54 and CD86 could be suppressed by dexamethasone treatment on isolated iDCs (CD54: 1.3-fold; CD86: 2.1-fold) as well as on the tissue-integrated iDCs (CD54: 1.4-fold; CD86: 1.6-fold). In conclusion, we were able to integrate THP-1-derived iDCs as functional dermal dendritic cell surrogates allowing the qualitative identification of potential sensitizers on the one hand, and drug candidates that potentially suppress sensitization on the other hand in a 3D human skin model corresponding to the 3R principles ("replace", "reduce" and "refine").

Keywords: CD86; DNCB; NF-κB; dermal dendritic cell; full-thickness skin model; nickel; p38 MAPK.

Publication types

Research Support, Non-U.S. Gov't
Comment

MeSH terms

Cytokines / metabolism
Dendritic Cells
Dinitrochlorobenzene* / pharmacology
Humans
Interleukin-6 / metabolism
Interleukin-8* / metabolism
Langerhans Cells

Substances

Dinitrochlorobenzene
Interleukin-8
Interleukin-6
Cytokines

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by a grant to NT, TB and KM by the Bundesministerium für Ernährung und Landwirtschaft (BMEL), funding number: 281A308C18.