Development of a co-culture of keratinocytes and immune cells for in vitro investigation of cutaneous sulfur mustard toxicity.
Identifieur interne : 000382 ( Main/Exploration ); précédent : 000381; suivant : 000383Development of a co-culture of keratinocytes and immune cells for in vitro investigation of cutaneous sulfur mustard toxicity.
Auteurs : Frank Balszuweit [Allemagne] ; Georg Menacher [Allemagne] ; Brunhilde Bloemeke [Allemagne] ; Annette Schmidt [Allemagne] ; Franz Worek [Allemagne] ; Horst Thiermann [Allemagne] ; Dirk Steinritz [Allemagne]Source :
- Chemico-biological interactions [ 1872-7786 ] ; 2014.
English descriptors
- KwdEn :
- Apoptosis (drug effects), Cell Line, Chemical Warfare Agents (toxicity), Coculture Techniques, Humans, Inflammation (chemically induced), Inflammation (immunology), Inflammation (pathology), Interleukin-6 (biosynthesis), Keratinocytes (drug effects), Keratinocytes (immunology), Keratinocytes (pathology), Langerhans Cells (drug effects), Langerhans Cells (immunology), Langerhans Cells (pathology), Monocytes (drug effects), Monocytes (immunology), Monocytes (pathology), Mustard Gas (toxicity), Necrosis.
- MESH :
- chemical , biosynthesis : Interleukin-6.
- chemical , toxicity : Chemical Warfare Agents, Mustard Gas.
- chemically induced : Inflammation.
- drug effects : Apoptosis, Keratinocytes, Langerhans Cells, Monocytes.
- immunology : Inflammation, Keratinocytes, Langerhans Cells, Monocytes.
- pathology : Inflammation, Keratinocytes, Langerhans Cells, Monocytes.
- Cell Line, Coculture Techniques, Humans, Necrosis.
Abstract
Sulfur mustard (SM) is a chemical warfare agent causing skin blistering, ulceration and delayed wound healing. Inflammation and extrinsic apoptosis are known to have an important role in SM-induced cytotoxicity. As immune cells are involved in those processes, they may significantly modulate SM toxicity, but the extent of those effects is unknown. We adapted a co-culture model of immortalized keratinocytes (HaCaT) and immune cells (THP-1) and exposed this model to SM. Changes in necrosis, apoptosis and inflammation, depending on SM challenge, absence or presence and number of THP-1 cells were investigated. THP-1 were co-cultured for 24h prior to SM exposure in order to model SM effects on immune cells continuously present in the skin. Our results indicate that the presence of THP-1 strongly increased necrosis, apoptosis and inflammation. This effect was already significant when the ratio of THP-1 and HaCaT cells was similar to the ratio of Langerhans immune cells and keratinocytes in vivo. Any further increases in the number of THP-1 had only slight additional effects on SM-induced cytotoxicity. In order to assess the effects of immune cells migrating into skin areas damaged by SM, we added non-exposed THP-1 to SM-exposed HaCaT. Those THP-1 had only slight effects on SM-induced cytotoxicity. Notably, in HaCaT exposed to 300μM SM, necrosis and inflammation were slightly reduced by adding intact THP-1. This effect was dependent on the number of immune cells, steadily increasing with the number of unexposed THP-1 added. In summary, we have demonstrated that (a) the presented co-culture is a robust model to assess SM toxicity and can be used to test the efficacy of potential antidotes in vitro; (b) immune cells, damaged by SM strongly amplified cytotoxicity, (c) in contrast, unexposed THP-1 (simulating migration of immune cells into affected areas after exposure in vivo) had no pronounced adverse, but exhibited some protective effects. Thus, protecting immune cells from SM toxicity may help to reduce overall injury.
DOI: 10.1016/j.cbi.2014.09.002
PubMed: 25256379
Affiliations:
- Allemagne
- Bavière, District de Cologne, District de Haute-Bavière, Rhénanie-Palatinat, Rhénanie-du-Nord-Westphalie
- Cologne, Munich, Trèves (Allemagne)
- Université Louis-et-Maximilien de Munich, Université de Trèves
Links toward previous steps (curation, corpus...)
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Le document en format XML
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<term>Cell Line</term>
<term>Chemical Warfare Agents (toxicity)</term>
<term>Coculture Techniques</term>
<term>Humans</term>
<term>Inflammation (chemically induced)</term>
<term>Inflammation (immunology)</term>
<term>Inflammation (pathology)</term>
<term>Interleukin-6 (biosynthesis)</term>
<term>Keratinocytes (drug effects)</term>
<term>Keratinocytes (immunology)</term>
<term>Keratinocytes (pathology)</term>
<term>Langerhans Cells (drug effects)</term>
<term>Langerhans Cells (immunology)</term>
<term>Langerhans Cells (pathology)</term>
<term>Monocytes (drug effects)</term>
<term>Monocytes (immunology)</term>
<term>Monocytes (pathology)</term>
<term>Mustard Gas (toxicity)</term>
<term>Necrosis</term>
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<keywords scheme="MESH" type="chemical" qualifier="toxicity" xml:lang="en"><term>Chemical Warfare Agents</term>
<term>Mustard Gas</term>
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<term>Monocytes</term>
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<front><div type="abstract" xml:lang="en">Sulfur mustard (SM) is a chemical warfare agent causing skin blistering, ulceration and delayed wound healing. Inflammation and extrinsic apoptosis are known to have an important role in SM-induced cytotoxicity. As immune cells are involved in those processes, they may significantly modulate SM toxicity, but the extent of those effects is unknown. We adapted a co-culture model of immortalized keratinocytes (HaCaT) and immune cells (THP-1) and exposed this model to SM. Changes in necrosis, apoptosis and inflammation, depending on SM challenge, absence or presence and number of THP-1 cells were investigated. THP-1 were co-cultured for 24h prior to SM exposure in order to model SM effects on immune cells continuously present in the skin. Our results indicate that the presence of THP-1 strongly increased necrosis, apoptosis and inflammation. This effect was already significant when the ratio of THP-1 and HaCaT cells was similar to the ratio of Langerhans immune cells and keratinocytes in vivo. Any further increases in the number of THP-1 had only slight additional effects on SM-induced cytotoxicity. In order to assess the effects of immune cells migrating into skin areas damaged by SM, we added non-exposed THP-1 to SM-exposed HaCaT. Those THP-1 had only slight effects on SM-induced cytotoxicity. Notably, in HaCaT exposed to 300μM SM, necrosis and inflammation were slightly reduced by adding intact THP-1. This effect was dependent on the number of immune cells, steadily increasing with the number of unexposed THP-1 added. In summary, we have demonstrated that (a) the presented co-culture is a robust model to assess SM toxicity and can be used to test the efficacy of potential antidotes in vitro; (b) immune cells, damaged by SM strongly amplified cytotoxicity, (c) in contrast, unexposed THP-1 (simulating migration of immune cells into affected areas after exposure in vivo) had no pronounced adverse, but exhibited some protective effects. Thus, protecting immune cells from SM toxicity may help to reduce overall injury.</div>
</front>
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