Cloning, expression, and functional characterization of rat MIP-2: a neutrophil chemoattractant and epithelial cell mitogen

• Published in: Journal of leukocyte biology Vol. 58; no. 3; pp. 359 - 364
• Main Authors: Driscoll, Kevin E., Hassenbein, Diana G., Howard, Brian W., Isfort, Robert J., Cody, David, Tindal, Michael H., Suchanek, Maureen, Carter, Janet M.
• Format: Journal Article
• Language: English
• Published: United States Society for Leukocyte Biology 01.09.1995
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cell Division - drug effects; chemokine; Chemokine CXCL2; Chemotaxis, Leukocyte - drug effects; CINC; Cloning, Molecular; Cytokines - genetics; Cytokines - pharmacology; DNA Primers - chemistry; DNA, Complementary - genetics; Epithelial Cells; Epithelium - drug effects; growth factor; Lung - cytology; Macrophages, Alveolar - cytology; MIP‐1α; MIP‐1β; MIP‐2; Mitogens; Molecular Sequence Data; Monokines - genetics; Monokines - pharmacology; Neutrophils - cytology; Neutrophils - drug effects; Rats; Rats, Inbred F344
• ISSN: 0741-5400; 1938-3673
• DOI: 10.1002/jlb.58.3.359

Macrophage inflammatory protein‐2 (MIP‐2) is a member of a family of cytokines that play roles in inflammatory, immune, and wound healing responses. To clone the cDNA for rat MIP‐2, RNA was isolated from the lungs of Fischer 344 rats after instillation of lipopolysaccharide. Reverse transcription‐polymerase chain reaction was performed by using synthetic oligonucleotide primers designed from the mouse MIP‐2 cDNA sequence. A cDNA containing the coding region of rat MIP‐2 was cloned and sequenced. Comparison to the mouse MIP‐2 cDNA demonstrated 90.3% homology at the nucleotide level and 86% homology at the amino acid level. The rat MIP‐2 cDNA was expressed in Escherichia coli and protein evaluated for bioactivity. The recombinant rat MIP‐2 was chemotactic for rat neutrophils but did not stimulate migration of rat alveolar macrophages or human peripheral blood eosinophils or lymphocytes. In addition, the recombinant rat MIP‐2 and the related rat chemokine, KC/CINC stimulated proliferation of rat alveolar epithelial cells but not fibroblasts in vitro.