Rainbow trout (Oncorhynchus mykiss) recombinant IL-1β and derived peptides induce migration of head-kidney leucocytes in vitro

The present work provides the first information concerning the chemoattractant activity of trout recombinant IL-1β and its derived peptides, referred to as P1, P2 and P3. The predicted rainbow trout mature interleukin-1β peptide was produced as a recombinant protein in Escherichia coli. The first peptide, P1, corresponded to fragment 146–157 (YVTPVPIETEAR) of the trout sequence and had an MW of 1·37 kDa. It was equivalent to a region known to be part of the receptor binding domain from the mammalian crystal structure of IL-1β complexed to its receptor. P2 was used as control peptide, consisting of the same 12 amino acids as P1, but arranged in a random sequence (VVEEYIRAPPTT). P3 was synthesised to complex with an adjacent region of the IL-1 receptor, and corresponded to fragment 207–216 (YRRNTGVDIS) of the trout sequence, with an MW of 1·18 kDa. Migration was stimulated when leucocytes were exposed to concentrations of ≥10 ng ml−1rIL-1β. Peptide P3 also induced leucocyte migration, with an optimal dose of 0·25 mm being recorded. While P1 had no effect on cell migration when used alone, synergism was evident as a consequence of combining P1 with a suboptimal dose (0·01 mm) of P3. No synergism occurred when cells were exposed to a combination of P3 and the control peptide P2.