Functional Characterization of <i>TNFα</i> in the Starry Flounder (<i>Platichthys stellatus</i>) and Its Potential as an Immunostimulant

<i>Tumor necrosis factor alpha</i> (<i>TNFα</i>) is a central pro-inflammatory cytokine that mediates host immune responses during infection. In this study, we identified and characterized the <i>TNFα</i> gene in the starry flounder (<i>Platichthys stellatus...

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Main Authors: Min-Young Sohn, Gyoungsik Kang, Kyung-Ho Kim, Ha-Jeong Son, Chan-Il Park
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Animals
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Online Access:https://www.mdpi.com/2076-2615/15/14/2119
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author Min-Young Sohn
Gyoungsik Kang
Kyung-Ho Kim
Ha-Jeong Son
Chan-Il Park
author_facet Min-Young Sohn
Gyoungsik Kang
Kyung-Ho Kim
Ha-Jeong Son
Chan-Il Park
author_sort Min-Young Sohn
collection DOAJ
description <i>Tumor necrosis factor alpha</i> (<i>TNFα</i>) is a central pro-inflammatory cytokine that mediates host immune responses during infection. In this study, we identified and characterized the <i>TNFα</i> gene in the starry flounder (<i>Platichthys stellatus</i>) through transcriptomic analysis. The deduced protein contained a conserved TNF domain and transmembrane region, and phylogenetic analysis confirmed its homology with other teleost TNFα proteins. Tissue-specific expression profiling revealed high baseline expression in immune-related peripheral organs and a distinct temporal modulation in response to <i>Streptococcus parauberis</i> infection. Recombinant TNFα (rTNFα), produced using a cell-free expression system, significantly enhanced phagocytic activity in peripheral and kidney-derived leukocytes in a dose-dependent manner. Peak activity was observed at 150–200 μg/mL, while a decline at higher concentrations suggested a threshold for immune stimulation. Importantly, hemolysis assays confirmed the safety of rTNFα even at the highest tested concentrations. These results demonstrate the immunomodulatory potential of TNFα as a molecular adjuvant in aquaculture vaccines and underscore its potential utility in immune-enhancing strategies for sustainable aquaculture.
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spelling doaj-art-ac4af63c759f46299145bccef87d0dab2025-08-20T03:13:42ZengMDPI AGAnimals2076-26152025-07-011514211910.3390/ani15142119Functional Characterization of <i>TNFα</i> in the Starry Flounder (<i>Platichthys stellatus</i>) and Its Potential as an ImmunostimulantMin-Young Sohn0Gyoungsik Kang1Kyung-Ho Kim2Ha-Jeong Son3Chan-Il Park4Department of Marine Biology & Aquaculture, College of Marine Science, Gyeongsang National University, Tongyeong 53064, Republic of KoreaDepartment of Aquatic Life Medicine, College of Marine Science, Gyeongsang National University, Tongyeong 53064, Republic of KoreaDepartment of Aquatic Life Medicine, College of Marine Science, Gyeongsang National University, Tongyeong 53064, Republic of KoreaDepartment of Marine Biology & Aquaculture, College of Marine Science, Gyeongsang National University, Tongyeong 53064, Republic of KoreaDepartment of Marine Biology & Aquaculture, College of Marine Science, Gyeongsang National University, Tongyeong 53064, Republic of Korea<i>Tumor necrosis factor alpha</i> (<i>TNFα</i>) is a central pro-inflammatory cytokine that mediates host immune responses during infection. In this study, we identified and characterized the <i>TNFα</i> gene in the starry flounder (<i>Platichthys stellatus</i>) through transcriptomic analysis. The deduced protein contained a conserved TNF domain and transmembrane region, and phylogenetic analysis confirmed its homology with other teleost TNFα proteins. Tissue-specific expression profiling revealed high baseline expression in immune-related peripheral organs and a distinct temporal modulation in response to <i>Streptococcus parauberis</i> infection. Recombinant TNFα (rTNFα), produced using a cell-free expression system, significantly enhanced phagocytic activity in peripheral and kidney-derived leukocytes in a dose-dependent manner. Peak activity was observed at 150–200 μg/mL, while a decline at higher concentrations suggested a threshold for immune stimulation. Importantly, hemolysis assays confirmed the safety of rTNFα even at the highest tested concentrations. These results demonstrate the immunomodulatory potential of TNFα as a molecular adjuvant in aquaculture vaccines and underscore its potential utility in immune-enhancing strategies for sustainable aquaculture.https://www.mdpi.com/2076-2615/15/14/2119<i>TNFα</i>molecular adjuvantstarry flounder
spellingShingle Min-Young Sohn
Gyoungsik Kang
Kyung-Ho Kim
Ha-Jeong Son
Chan-Il Park
Functional Characterization of <i>TNFα</i> in the Starry Flounder (<i>Platichthys stellatus</i>) and Its Potential as an Immunostimulant
Animals
<i>TNFα</i>
molecular adjuvant
starry flounder
title Functional Characterization of <i>TNFα</i> in the Starry Flounder (<i>Platichthys stellatus</i>) and Its Potential as an Immunostimulant
title_full Functional Characterization of <i>TNFα</i> in the Starry Flounder (<i>Platichthys stellatus</i>) and Its Potential as an Immunostimulant
title_fullStr Functional Characterization of <i>TNFα</i> in the Starry Flounder (<i>Platichthys stellatus</i>) and Its Potential as an Immunostimulant
title_full_unstemmed Functional Characterization of <i>TNFα</i> in the Starry Flounder (<i>Platichthys stellatus</i>) and Its Potential as an Immunostimulant
title_short Functional Characterization of <i>TNFα</i> in the Starry Flounder (<i>Platichthys stellatus</i>) and Its Potential as an Immunostimulant
title_sort functional characterization of i tnfα i in the starry flounder i platichthys stellatus i and its potential as an immunostimulant
topic <i>TNFα</i>
molecular adjuvant
starry flounder
url https://www.mdpi.com/2076-2615/15/14/2119
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