02886nas a2200325   4500000000100000000000100001008004100002260001500043653000900058653000900067653003000076653001100106653001100117653002600128653001300154100002600167700002000193700001800213700001800231700001800249700001400267700002700281700002000308700002800328245010100356856009300457490000700550520198900557022001402546       2023                            d  c2023-11-0610aCD8610aDNCB10aFull-thickness skin model10aNF-κB10aNickel10adermal dendritic cell10ap38 MAPK1 aJohanna Maria Hölken1 aKatja Friedrich1 aMarion Merkel1 aNelli Blasius1 aUrsula Engels1 aTimo Buhl1 aKarsten Rüdiger Mewes1 aLars Vierkotten1 aNicole Elisabeth Teusch00aA human 3D immune competent full-thickness skin model mimicking dermal dendritic cell activation  uhttps://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2023.1276151/full0 v143 a<p>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<sup>-</sup>, CD11c<sup>+</sup> 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 NiSO<sub>4</sub> (1.6-fold). Furthermore, NiSO<sub>4</sub> 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 NiSO<sub>4</sub>: 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, NiSO<sub>4</sub> 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”).</p>  a1664-3224