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Cloning, expression, and functional characterization of rat MIP‐2: a neutrophil chemoattractant and epithelial cell mitogen
Author(s) -
Driscoll Kevin E.,
Hassenbein Diana G.,
Howard Brian W.,
Isfort Robert J.,
Cody David,
Tindal Michael H.,
Suchanek Maureen,
Carter Janet M.
Publication year - 1995
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.58.3.359
Subject(s) - complementary dna , biology , microbiology and biotechnology , chemokine , macrophage inflammatory protein , recombinant dna , lipopolysaccharide , chemotaxis , molecular cloning , gene , inflammation , immunology , biochemistry , receptor
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.