Highlights
- •Primary perilesional keratinocytes (PLK) obtained from vitiligo patients were used.
- •PLK were treated with low-dose 10 fg/ml IL-4, IL-10, bFGF and β-endorphin for 48 h.
- •IL-4, IL-10, bFGF and β-endorphin were prepared by sequential-kinetic-activation.
- •Treatment reduced extra- and intracellular ROS production, increased viability in PLK.
- •Low-dose IL-4, IL-10, bFGF and β-endorphin represent a therapeutic tool for vitiligo.
Abstract
Background
Vitiligo is a systemic dermatological disorder characterized by the loss of skin pigmentation
due to melanocyte injury or aberrant functioning. Recent data underline its multifactorial
etiology with significant involvement of autoimmune and redox alterations. The major
role in vitiligo cellular immunity is displayed by augmented Th1 and Th17 and suppressed
TREGs and Th2 lymphocyte populations. Our previous studies indicate a marked redox
imbalance in perilesional (“PL”, i.e. obtained from visibly unaffected skin surrounding
the depigmented area in vitiligo patients) keratinocytes where the massive infiltration
of inflammatory cells takes place. No defined therapy exists for vitiligo. Although
a number of approaches have been used for the induction of TREGs and Th2 cells, they
may be associated with significant off-target effects.
Objective
In order to identify a targeted approach for vitiligo treatment we, first, aimed to
investigate the possible source of ROS overproduction in PL keratinocytes. Second,
we tested the effect of low-dose selected cytokines, on intra- and extracellular ROS
production, cell viability and cell cycle of PL keratinocytes.
Methods
The in vitro study was conducted on primary PL keratinocytes obtained from the skin
of vitiligo patients in our previous studies. The activity of NADPH oxidase was measured
on intact PL and control keratinocytes, treated or not with cytokines, by luminometric
assay. The following cytokines were selected for PL keratinocytes treatment: IL-10
and IL-4 (produced by TREGs and Th2, respectively), basic fibroblasts growth factor
(bFGF) and neuropeptide β-endorphin (modulating the cellular resistance to oxidative
stress and the immune response, respectively). All cytokines were used at concentration
of 10 fg/ml and were prepared by sequential-kinetic-activation (SKA). Intracellular ROS
production and cell cycle were analyzed by flow cytometry using H2DCFDA and propidium
iodide dyes, respectively. Cell viability was measured by fluorometric resazurin reduction
method.
Results
Our results suggest that NADPH oxidase represents one of the main sources of ROS overproduction
by PL keratinocytes. Further, SKA low-dose IL-10, β-endorphin and, particularly, IL-4
and bFGF display a positive effect on redox dyshomeostasis and viability and, in our
experimental conditions, don’t affect the cell cycle of PL keratinocytes.
Conclusion
Our preliminary data suggest that low-dose IL-10, IL-4, β-endorphin and bFGF can be
proposed as a new therapeutic tool for vitiligo treatment.
Keywords
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Article info
Publication history
Published online: May 18, 2015
Accepted:
May 11,
2015
Received:
April 19,
2015
Identification
Copyright
© 2015 Japanese Society for Investigative Dermatology. Published by Elsevier Inc. All rights reserved.
