ArticleMolecular biology reports2026
Production and validation of recombinant hamster cytokines as tools for immunological studies.
Article in Molecular biology reports, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
backgroundCellular immunity studies in Syrian hamsters have been hampered by the lack of species-specific immunological reagents, particularly cytokines. To overcome this limitation, we characterized the biological activity of recombinant hamster cytokines IFN-γ, TGF-β, IL-6, IL-10 and TNF-α previously produced in Escherichia coli in a mammalian cell model (CHO-K1 cells).
methodsRecombinant proteins were purified, detoxified and their activity assessed by metabolic activity (MTT) and gene expression (RT-qPCR) assays in CHO-K1 cells. The suitability of this cell model for this purpose was also determined through RT-qPCR, validating the expression of cytokine receptors and associated signaling pathway markers.
resultsAll cytokines except rIFN-γ increased metabolic activity at 24 h, whereas rIFN-γ induced a dose-dependent reduction without evidence of cytotoxicity, suggesting a potential cell cycle arrest rather than cell death. At 48 h, all cytokines induced a decrease in metabolic activity. CHO-K1 cells expressed all protein receptor genes evaluated, supporting their relevance as a model to determine recombinant cytokine activity. Transcriptional analysis revealed pathway specific effects: rIFN-γ markedly induced STAT1; rTGF-β increased SMAD2/3 expression, and rTNF-α upregulated NFκB. Gene expression analysis also revealed that rIL-6 downregulated tnfα, while rIL-10 and rTGF-β reduced tnfα and ifnγ expression. rIFN-γ also suppressed tnfα, with limited effects on other genes. These responses were consistent with the expected signaling dynamics of each cytokine.
conclusionsRecombinant hamster cytokines exhibit functional activity in CHO-K1 cells, as shown by metabolic responses, modulation of immune-related genes, and activation of expected signaling mediators. Together with the detection of cytokine receptor transcripts, these results support the use of CHO-K1 cells for evaluating recombinant cytokine bioactivity and broaden the range of validated immunological tools available for Syrian hamster research.
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