IFN-γ/TNF-α Synergism Induces Pro-Inflammatory Cytokine and Chemokine Production by In Vitro Canine Keratinocytes
Activated keratinocytes play a crucial role in skin inflammation through the production of multiple inflammatory mediators; however, little is known about cytokine secretion by activated keratinocytes in dogs. This study aimed to investigate the effects of the Th1 and Th2 types of cytokines on the p...
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2025-01-01
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| Series: | Veterinary Sciences |
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| author | Kyungsook Jung Ji-Yeong Ku Je-Seong Kwon Gayeon Won Hakyoung Yoon Sang-Ik Oh Mi Hye Kim Chongchan Kim Ji-Seon Yoon |
| author_facet | Kyungsook Jung Ji-Yeong Ku Je-Seong Kwon Gayeon Won Hakyoung Yoon Sang-Ik Oh Mi Hye Kim Chongchan Kim Ji-Seon Yoon |
| author_sort | Kyungsook Jung |
| collection | DOAJ |
| description | Activated keratinocytes play a crucial role in skin inflammation through the production of multiple inflammatory mediators; however, little is known about cytokine secretion by activated keratinocytes in dogs. This study aimed to investigate the effects of the Th1 and Th2 types of cytokines on the production of keratinocyte-derived inflammatory mediators. Canine progenitor epidermal keratinocytes (CPEKs) were incubated with canine recombinant IL-4, IL-13, an IL4/IL13 mixture, IFN-γ, TNF-α, or an IFN-γ/TNF-α mixture for 24 h following 100% confluency. Culture supernatants were analyzed for cytokine concentration, including chemokine ligand (CXCL) 8, IL-10, IL-6, IL-1ß, IL-12, and chemokine ligand 2 (CCL2) by enzyme-linked immunosorbent assay. CPEKs incubated with the IFN-γ/TNF-α mixture showed significantly increased IL-6 concentration. In addition, significantly increased concentrations of CXCL8 were detected in CPEKs incubated with TNF-α and with the IFN-γ/TNF-α mixture. CCL2 concentrations increased in cells incubated with IFN-γ, TNF-α, and the IFN-γ/TNF-α mixture. The IFN-γ/TNF-α mixture synergistically enhanced CCL2 production. Dose-dependent elevations were also observed in IL-6 in response to the IFN-γ/TNF-α mixture, and in CCL2 in response to IFN-γ, TNF-α, and the IFN-γ/TNF-α mixture. These findings indicate that IFN-γ and TNF-α synergistically increase pro-inflammatory cytokines and chemokines secreted by canine keratinocytes. This in vitro culture system could be useful to investigate cytokine-mediated crosstalk between keratinocytes and immune cells and new therapeutic strategies for keratinocyte-mediated inflammatory skin diseases. |
| format | Article |
| id | doaj-art-fa497d960c2d46318a14bbcdbdf07490 |
| institution | Kabale University |
| issn | 2306-7381 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj-art-fa497d960c2d46318a14bbcdbdf074902025-01-24T13:52:08ZengMDPI AGVeterinary Sciences2306-73812025-01-011215510.3390/vetsci12010055IFN-γ/TNF-α Synergism Induces Pro-Inflammatory Cytokine and Chemokine Production by In Vitro Canine KeratinocytesKyungsook Jung0Ji-Yeong Ku1Je-Seong Kwon2Gayeon Won3Hakyoung Yoon4Sang-Ik Oh5Mi Hye Kim6Chongchan Kim7Ji-Seon Yoon8Functional Biomaterials Research Center, Korea Research Institute of Bioscience and Biotechnology, Jeongeup 56212, Republic of KoreaBiosafety Research Institute and College of Veterinary Medicine, Jeonbuk National University, Iksan 54896, Republic of KoreaBiosafety Research Institute and College of Veterinary Medicine, Jeonbuk National University, Iksan 54896, Republic of KoreaBiosafety Research Institute and College of Veterinary Medicine, Jeonbuk National University, Iksan 54896, Republic of KoreaBiosafety Research Institute and College of Veterinary Medicine, Jeonbuk National University, Iksan 54896, Republic of KoreaBiosafety Research Institute and College of Veterinary Medicine, Jeonbuk National University, Iksan 54896, Republic of KoreaCollege of Korean Medicine, Woosuk University, Wanju 55338, Republic of KoreaKorea Thumb Vet Co., Ltd., 470-15 Seonhwa-ro, Iksan 54631, Republic of KoreaBiosafety Research Institute and College of Veterinary Medicine, Jeonbuk National University, Iksan 54896, Republic of KoreaActivated keratinocytes play a crucial role in skin inflammation through the production of multiple inflammatory mediators; however, little is known about cytokine secretion by activated keratinocytes in dogs. This study aimed to investigate the effects of the Th1 and Th2 types of cytokines on the production of keratinocyte-derived inflammatory mediators. Canine progenitor epidermal keratinocytes (CPEKs) were incubated with canine recombinant IL-4, IL-13, an IL4/IL13 mixture, IFN-γ, TNF-α, or an IFN-γ/TNF-α mixture for 24 h following 100% confluency. Culture supernatants were analyzed for cytokine concentration, including chemokine ligand (CXCL) 8, IL-10, IL-6, IL-1ß, IL-12, and chemokine ligand 2 (CCL2) by enzyme-linked immunosorbent assay. CPEKs incubated with the IFN-γ/TNF-α mixture showed significantly increased IL-6 concentration. In addition, significantly increased concentrations of CXCL8 were detected in CPEKs incubated with TNF-α and with the IFN-γ/TNF-α mixture. CCL2 concentrations increased in cells incubated with IFN-γ, TNF-α, and the IFN-γ/TNF-α mixture. The IFN-γ/TNF-α mixture synergistically enhanced CCL2 production. Dose-dependent elevations were also observed in IL-6 in response to the IFN-γ/TNF-α mixture, and in CCL2 in response to IFN-γ, TNF-α, and the IFN-γ/TNF-α mixture. These findings indicate that IFN-γ and TNF-α synergistically increase pro-inflammatory cytokines and chemokines secreted by canine keratinocytes. This in vitro culture system could be useful to investigate cytokine-mediated crosstalk between keratinocytes and immune cells and new therapeutic strategies for keratinocyte-mediated inflammatory skin diseases.https://www.mdpi.com/2306-7381/12/1/55caninekeratinocyteinterferon γtumor necrosis factor αsynergistic effectspro-inflammatory cytokine |
| spellingShingle | Kyungsook Jung Ji-Yeong Ku Je-Seong Kwon Gayeon Won Hakyoung Yoon Sang-Ik Oh Mi Hye Kim Chongchan Kim Ji-Seon Yoon IFN-γ/TNF-α Synergism Induces Pro-Inflammatory Cytokine and Chemokine Production by In Vitro Canine Keratinocytes Veterinary Sciences canine keratinocyte interferon γ tumor necrosis factor α synergistic effects pro-inflammatory cytokine |
| title | IFN-γ/TNF-α Synergism Induces Pro-Inflammatory Cytokine and Chemokine Production by In Vitro Canine Keratinocytes |
| title_full | IFN-γ/TNF-α Synergism Induces Pro-Inflammatory Cytokine and Chemokine Production by In Vitro Canine Keratinocytes |
| title_fullStr | IFN-γ/TNF-α Synergism Induces Pro-Inflammatory Cytokine and Chemokine Production by In Vitro Canine Keratinocytes |
| title_full_unstemmed | IFN-γ/TNF-α Synergism Induces Pro-Inflammatory Cytokine and Chemokine Production by In Vitro Canine Keratinocytes |
| title_short | IFN-γ/TNF-α Synergism Induces Pro-Inflammatory Cytokine and Chemokine Production by In Vitro Canine Keratinocytes |
| title_sort | ifn γ tnf α synergism induces pro inflammatory cytokine and chemokine production by in vitro canine keratinocytes |
| topic | canine keratinocyte interferon γ tumor necrosis factor α synergistic effects pro-inflammatory cytokine |
| url | https://www.mdpi.com/2306-7381/12/1/55 |
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