Expression of Interleukin-1β protein in vitro, ex vivo and in vivo salmonid models
Interleukin-1β (IL-1β) is one of the first cytokines expressed during immune responses, and its levels are affected by many factors, including stress. To date, it has only been possible to measure IL-1β transcript (mRNA) expression quantitatively in fish using qPCR. This is because previous studies...
| Autores: | , , , , , , , , , , , , , |
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| Tipo de documento: | artigo |
| Estado: | Versão publicada |
| Data de publicação: | 2023 |
| País: | Brasil |
| Recursos: | Universidade Estadual Paulista (UNESP) |
| Repositório: | Repositório Institucional da UNESP |
| Idioma: | inglês |
| OAI Identifier: | oai:repositorio.unesp.br:11449/300974 |
| Acesso em linha: | http://dx.doi.org/10.1016/j.dci.2023.104767 https://hdl.handle.net/11449/300974 |
| Access Level: | Acceso aberto |
| Palavra-chave: | Cell culture Diagnostic assay Immunostimulation Interleukin-1β Natural infection Quantitative ELISA |
| Resumo: | Interleukin-1β (IL-1β) is one of the first cytokines expressed during immune responses, and its levels are affected by many factors, including stress. To date, it has only been possible to measure IL-1β transcript (mRNA) expression quantitatively in fish using qPCR. This is because previous studies that measured IL-1β protein concentrations in these taxa used western blotting, which only provides qualitative data. To advance our knowledge of fish IL-1β biology, and because post-translational processing plays a critical role in the activation of this molecule, we developed a quantitative enzyme-linked immunosorbent assay (ELISA) to accurately measure the concentration of IL-1β protein in several cell cultures and in vivo in salmonids. We compared changes in IL-1β protein levels to the expression of its mRNA. The developed ELISA was quite sensitive and has a detection limit of 12.5 pg/mL. The tools developed, and information generated through this research, will allow for a more accurate and complete understanding of IL-1β′s role in the immune response of salmonids.The assay described here has the potential to significantly advance our ability to assess fish health and immune status. |
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