Recombinant Production and Biological Properties of Rat Cytokine‐Induced Neutrophil Chemoattractants, GRO/CINC‐2α, CINC‐2β and CINC‐3

• Published in: European journal of biochemistry Vol. 231; no. 2; pp. 306 - 311
• Main Authors: Shibata, Futoshi, Konishi, Kiyoshi, Kato, Hideko, Komorita, Naruyasu, Al‐Mokdad, Maher, Fujioka, Motoji, Nakagawa, Hideo
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 15-07-1995
• Subjects: Animals; Base Sequence; Calcium - metabolism; Cathepsin G; Cathepsins - metabolism; Chemokine CXCL1; Chemokine CXCL2; Chemokines, CXC; Chemotactic factor; Chemotactic Factors - biosynthesis; Chemotactic Factors - genetics; Chemotactic Factors - pharmacology; Chemotaxis, Leukocyte; Cloning, Molecular; Cytokines - biosynthesis; Cytokines - genetics; Cytokines - pharmacology; cytokine‐induced neutrophil chemoattractant; DNA, Complementary - genetics; Escherichia coli - genetics; Gene Expression; GRO chemoattractant; Growth Substances - biosynthesis; Growth Substances - genetics; Growth Substances - pharmacology; Intercellular Signaling Peptides and Proteins; Lipopolysaccharides - pharmacology; macrophage inflammatory protein 2; Macrophages, Peritoneal - chemistry; Male; Molecular Sequence Data; Monokines - biosynthesis; Monokines - genetics; Monokines - pharmacology; Neutrophils - drug effects; Neutrophils - physiology; Polymerase Chain Reaction; Rats; Rats, Wistar; recombinant expression; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Recombinant Proteins - pharmacology; Sequence Analysis; Serine Endopeptidases
• DOI: 10.1111/j.1432-1033.1995.0306e.x
• EISSN: 1432-1033
• ISSN: 0014-2956

Recently we found four cytokine‐induced neutrophil chemoattractants, CINC‐1, CINC‐2α, CINC‐2β and CINC‐3/macrophage inflammatory protein 2 (MIP‐2), in conditioned medium of granulation tissue obtained from carrageenin‐induced inflammation in rats [Nakagawa, H., Komorita, N., Shibata, E, Ikesue, A., Konishi, K., Fujioka, M. & Kato, H. (1994) Biochem. J. 301, 545–550]. In the present report, we describe recombinant production of CINC‐2α, CINC‐2β and CINC‐3 in Escherichia coli, and biological properties of these chemokines. Neutrophil chemotactic activities of CINC‐2α and 2βin vitro 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 neutrophils with CINCs induced an increase in intracellular [Ca2+] dose‐dependently. CINC‐3 was more potent than the other CINCs and still induced an increase in intracellular [Ca2+] in rat neutrophils stimulated first with other CINCs. CINC‐2α, CINC‐2β and CINC‐3 induced a comparable response to CINC‐1 in the release of cathepsin G from rat neutrophils. Injection of CINC‐2α, 2β 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‐2α, 2β and 3 as well as CINC‐1 are chemoattractants specific for neutrophil in vivo.