RECOMBINANT PRODUCTION AND BIOLOGICAL PROPERTIES OF RAT CYTOKINE-INDUCED NEUTROPHIL CHEMOATTRACTANTS, GRO CINC-2-ALPHA, CINC-2-BETA AND CINC-3/

Recently we found four cytokine-induced neutrophil chemoattractants, C INC-1, CINC-2 alpha, CINC-2 beta and CINC-3/macrophage inflammatory pr otein 2 (MIP-2), in conditioned medium of granulation tissue obtained from carrageenin-induced inflammation in rats [Nakagawa, H., Komorita, N., Shibata, F. Ikesue, A., Konishi, K., Fujioka, M. and Kato, H. (19 94) Biochem. J. 301, 545-550]. In the present report, we describe reco mbinant production of CINC-2 alpha, CINC-2 beta and CINC-3 in Escheric hia coli, and biological properties of these chemokines. Neutrophil ch emotactic activities of CINC-2 alpha and 2 beta in vibro were the same as the activity of CINC-1. CINC-3 had an activity comparable to other CINCs, but showed a decrease at high concentrations. Stimulation of n eutrophils with CINCs induced an increase in intracellular [Ca2+] dose -dependently. CINC-3 was more potent than the other CINCs and still in duced an increase in intracellular [Ca2+] in rat neutrophils stimulate d first with other CINCs. CINC-2 alpha, CINC-2 beta and CINC-3 induced a comparable response to CINC-1 in the release of cathepsin G from ra t neutrophils. Injection of CINC-2 alpha, 2 beta and 3 into preformed air-pouch on the back of rat induced infiltration of neutrophils to an extent similar to that caused by the injection of CINC-1. These data indicate CINC-2 alpha, 2 beta and 3 as well as CINC-1 are chemoattract ants specific for neutrophil in vivo.