Identification of mouse granulocyte chemotactic protein-2 from fibroblasts and epithelial cells. Functional comparison with natural KC and macrophage inflammatory protein-2.
Neutrophil and monocyte chemotactic factors were isolated from conditioned media of mouse fibroblasts and epithelial cells. Neutrophil chemotactic activities were purified to homogeneity using a four-step chromatographic procedure, and the corresponding proteins were identified by amino acid sequence analysis. Natural forms of the murine chemokines KC and macrophage inflammatory protein-2 were isolated from virus-infected fibroblasts. However, the major neutrophil chemotactic activity from fibroblasts stimulated with endotoxin plus double-stranded RNA and from PMA-treated epithelial cells resided in other 7- and 8-kDa proteins. Amino acid sequence analysis revealed a novel Cys-Xaa-Cys chemokine structure, characterized by the conservation of four cysteines and the Glu-Leu-Arg motif. Based on the completely identified primary structure of this natural protein, this chemokine must be considered to be the murine homologue of human and bovine granulocyte chemotactic protein-2 (GCP-2; 61 and 64% identical residues, respectively). Due to NH2-terminal cleavage, 11 different forms of mouse GCP-2 were discovered. In contrast to human and bovine GCP-2, functional comparison of long and short NH2-terminal forms of mouse GCP-2 demonstrated that truncated mouse GCP-2 (short form) has a higher specific activity in neutrophil activation (gelatinase B release) and chemotaxis assays. Furthermore, mouse GCP-2 was more potent than human GCP-2 on human neutrophils, and more active than murine KC and macrophage inflammatory protein-2 on mouse neutrophils. In view of the absence of a murine homologue for IL-8, NH2-terminally processed GCP-2 can be considered a major neutrophil chemoattractant in the mouse during the inflammatory response.
Wuyts
A
A
Rega Institute for Medical Research, Laboratory of Molecular Immunology, University of Leuven, Belgium.
Haelens
A
A
Proost
P
P
Lenaerts
J P
JP
Conings
R
R
Opdenakker
G
G
Van Damme
J
J
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D054360:Chemokine CXCL1; D054426:Chemokine CXCL2; D054427:Chemokine CXCL6; D018925:Chemokines / Q000302:isolation & purification* / Q000494:pharmacology / Q000502:physiology; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000494:pharmacology; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000737:chemistry; D016207:Cytokines / Q000302:isolation & purification / Q000502:physiology; D004847:Epithelial Cells; D004848:Epithelium / Q000737:chemistry* / Q000187:drug effects; D005347:Fibroblasts / Q000737:chemistry* / Q000187:drug effects / Q000821:virology; D006801:Humans; D015850:Interleukin-6 / Q000032:analysis; D051379:Mice; D008969:Molecular Sequence Data; D015846:Monokines / Q000302:isolation & purification / Q000502:physiology; D010222:Parainfluenza Virus 1, Human / Q000502:physiology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
C515503:CXCL6 protein, human; D054360:Chemokine CXCL1; D054426:Chemokine CXCL2; D054427:Chemokine CXCL6; D018925:Chemokines; D019743:Chemokines, CXC; D002630:Chemotactic Factors; D017077:Culture Media, Conditioned; C475895:Cxcl1 protein, mouse; D016207:Cytokines; D015850:Interleukin-6; D015846:Monokines; C074302:keratinocyte-derived chemokines; D013755:Tetradecanoylphorbol Acetate