Analysis of genes isolated from lipopolysaccharide-stimulated rainbow trout (Oncorhynchus mykiss) macrophages



Goetz FW, Iliev DB, McCauley LAR, Liarte CQ, Tort L, Planas JV & MacKenzie S (2004) Analysis of genes isolated from lipopolysaccharide-stimulated rainbow trout (Oncorhynchus mykiss) macrophages. Molecular Immunology, 41 (12), pp. 1199-1210.

A primary cell culture system was used to obtain differentiated rainbow trout (Oncorhynchus mykiss) macrophages that were stimulated with Escherichia coli lipopolysaccharide (LPS-10μg/ml) for 12h in vitro. Messenger RNA from the LPS-stimulated cells was used to create two cDNA libraries from which a total of 1048 sequences were analyzed. A large number of cDNAs were obtained that could be related to immune function including structural proteins, proteases and antiproteases, regulators of transcription and translation, cell death regulators, receptors, lectins and immunoglobulins, cytokines and chemokines, cell surface antigens, signal transduction proteins, antimicrobial peptides, and enzymes involved in eicosanoid synthesis. Selected genes that were analyzed by RT-PCR and real time PCR and found to be upregulated by LPS, included vascular cell adhesion molecule, the CCAAT/enhancer binding protein β, the inhibitor of NF-kB α, CD209, a major histocompatibility class II-invariant chain protein, cyclin L1, acute phase serum amyloid A, and prostaglandin endoperoxide synthase 2.

Rainbow trout; Macrophage; Lipopolysaccharide; Head kidney; Transcriptomics; cDNA libraries

Molecular Immunology: Volume 41, Issue 12

Publication date30/11/2004

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Professor Simon MacKenzie

Professor Simon MacKenzie

Professor & Head of Inst of Aquaculture, Institute of Aquaculture