The production and bioactivity of rainbow trout (Oncorhynchus mykiss) recombinant IL-1β

Identification and expression analysis of an atypical chemokine receptor-2 (ACKR2)/CC chemokine binding protein-2 (CCBP2) in rainbow trout (Oncorhynchus mykiss)

“…Four day old primary HK macrophages were prepared as described previously [40] and stimulated with a variety of PAMPS and cytokines, including polyinosinic: polycytidylic acid (poly I:C, 50 mg/ml, Sigma), peptidoglycan (PGN, 5 mg/ml, Invivogen), rIL-6 (100 ng/ml) [40], rIFN-g (20 ng/ml) [41], rTNF-a (10 ng/ml) [42], rIL-1b (20 ng/ml) [43], and two rIL-12 isoforms that differ in the p40 chain [44] for 4 h, 8 h and 24 h. As above, the concentration chosen for each stimulant was deemed optimal for immune gene expression experiments based on our previous studies [40e44]. Real-time PCR quantification of expression was as described above and the results were expressed as a fold change relative to the time-matched unstimulated controls.…”

Cloning of two rainbow trout nucleotide-binding oligomerization domain containing 2 (NOD2) splice variants and functional characterization of the NOD2 effector domains

“…The cells were passaged into fresh culture flasks 2 days before being stimulated for 6 h with either 20 ng/ml recombinant trout IL-1b (rIL-1b) protein [22], or 20 ng/ml recombinant trout interferon-g protein (rIFN-g) [23] or left untreated as control. The RNA preparation and real-time PCR analysis were as described above.…”

Recombinant interleukin-1 beta activates the hypothalamic-pituitary-interrenal axis in rainbow trout, Oncorhynchus mykiss

“…Trout rIL-1 was purified using a modified protocol from the method described by Hong et al (2001). The six histidine-tagged cDNA sequence encoding the predicted mature IL-1 peptide was cloned into the vector pQE30 (Invitrogen) and transformed into E. coli cells (JM109; Promega).…”