Evaluation of blood cell viability rate, gene expression, and O-GlcNAcylation profiles as indicative signatures for fungal stimulation of salmonid cell models. (February 2022)

• Record Type: Journal Article
• Authors: Magray, Aqib Rehman; Ribera, Joan Martorell; Isernhagen, Lisa; Galuska, Sebastian P.; Günther, Juliane; Verleih, Marieke; Viergutz, Torsten; Brunner, Ronald M.; Ganai, Bashir Ahmad; Ahmad, Fayaz; Zlatina, Kristina; Rebl, Alexander
• Is Part Of: Molecular immunology. Volume 142(2022)
• Page Start: 120
• Page End: 129
• Publication Date: 2022-02
• Subjects: Fungi -- Innate immune response -- Pattern-recognition receptors -- Teleost; Immunochemistry -- Periodicals; Molecular biology -- Periodicals; Immunochemistry -- Periodicals; Allergy and Immunology -- Periodicals; Molecular Biology -- Periodicals; Immunochimie -- Périodiques; Biologie moléculaire -- Périodiques; Immunochemistry; Molecular biology; Periodicals; Electronic journals; 571.96
• DOI: 10.1016/j.molimm.2021.12.019
• Languages: English
• ISSNs: 0161-5890

Abstract

Highlights: We analysed the response of rainbow trout to Mucor, Fusarium and fungal stimulants. Mucor treatment induced the most pronounced transcriptional changes. Mucor treatment slightly modulated the cellular O -GlcNAcylation status. Fungal stimulants induced death of blood cells to a greater extent than whole fungi. Abstract: Fungal diseases of fish are a significant economic problem in aquaculture. Using high-throughput expression analysis, we identified potential transcript markers in primary head kidney and secondary embryonic cells from salmonid fish after stimulation with the inactivated fungi Mucor hiemalis and Fusarium aveneacium and with purified fungal molecular patterns. The transcript levels of most of the 45 selected genes were altered in head-kidney cells after 24 h of stimulation with fungal antigens. Stimulation with the inactivated fungus M. hiemalis induced the most pronounced transcriptional changes, including the pathogen receptor-encoding genes CLEC18A and TLR22, the cytokine-encoding genes IL6 and TNF, and the gene encoding the antimicrobial peptide LEAP2 . In parallel, we analyzed the total GlcNAcylation status of embryonic salmonid cells with or without stimulation with inactivated fungi. O -GlcNAcylation modulates gene expression, intracellular protein, and signal activity, but we detected no significant differences after a 3-h stimulation. A pathway analysis tool identified the "apoptosis of leukocytes" based on the expression profile 24 h after fungal stimulation. Fluorescence microscopy combined with flow cytometry revealed apoptosis in 50 % of head-kidney leukocytes after 3 h stimulation with M. hiemalis, but this level decreased by > 5% after 24 h of stimulation. The number of apoptotic cells significantly increased in all blood cells after a 3-h stimulation with fungal molecular patterns compared to unstimulated controls. This in vitro approach identified transcript-based parameters that were strongly modulated by fungal infections of salmonid fish.