Science and Technology Production
Congress
Authorship
Campo V
;
PAZ, MARIELA LAURA
;
Leoni J
;
González Maglio DH
;
Cela EM
Date
2018
Publishing House and Editing Place
Estudio Sigma SRL
Summary
Information provided by the agent in
SIGEVA
Cutaneous immune system comprises a complex network of cells, including keratinocytes (in the epidermis) and macrophages (in the dermis). Skin UVB irradiation affects mainly the epidermis, leading keratinocytes to release a wide variety of cytokines and soluble mediators that are significantly involved in immunoregulation. The aim of the present work was to evaluate the influence of UV-induced keratinocytes? soluble mediators on macrophages response to a bacterial stimulus.Human keratinocytes (...
Cutaneous immune system comprises a complex network of cells, including keratinocytes (in the epidermis) and macrophages (in the dermis). Skin UVB irradiation affects mainly the epidermis, leading keratinocytes to release a wide variety of cytokines and soluble mediators that are significantly involved in immunoregulation. The aim of the present work was to evaluate the influence of UV-induced keratinocytes? soluble mediators on macrophages response to a bacterial stimulus.Human keratinocytes (HaCaT cells) were exposed to UVB (0, 12.5, 25 and 50 mJ/cm2) and cultured for 24h. HaCaT supernatants (HSN) were collected, TNF-α, IL-6, IL-1β and IL-10 production was measured (ELISA) and, finally, HSN were used to condition macrophage response. Human monocytic cell line (THP-1) was differentiated to macrophages (PMA), challenged with 0, 50, 100 and 200 µg/ml of heat-killed BCG, and cultured for 24h to measured cytokines production. Subsequently, THP-1 macrophages were challenged in the presence of HSN, and cytokine and ROS production (DCF-DA) were evaluated 24 and 2h after the challenge, respectively.UVB-irradiated keratinocytes increased the production of the evaluated cytokines, reaching the maximum levels with a 25mJ/cm2 exposure: TNF-α, IL-6, IL-1β and IL-10 increased 2.5, 20, 2.5 and 2 times vs. non-irradiated controls, respectively. Cells exposed to 50mJ/cm2 produced lower cytokines levels (2, 1.5, 1.5 and 2 fold decrease, respectively). BCG stimulation of THP-1 cells did not change IL-6 and IL-10 production, but increased IL-1β (50, 61 and 80 vs. 29pg/ml) and TNF-α production (162 vs. 65pg/ml). HSN did not affect TNF-α, IL-6 and IL-10 production by stimulated THP-1 cells, but significantly increased IL-1β production (50 and 100µg/ml BCG: p<0.001, p<0.01, respectively). Moreover, both BCG concentration significantly increased THP-1 oxidative metabolism in irradiated and non-irradiated cells (p<0.001, p<0.01 respectively).These results suggest that UVB radiation can modulate keratinocytes? cytokine production, conditioning the macrophages response (ROS and cytokine production) to microbial challenges.
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Key Words
INNATE RESPONSEMACROPHAGESKERATINOCYTESUV RADIATION