Recombinant mouse granulocyte chemotactic protein-2: production in bacteria, characterization, and systemic effects on leukocytes.
Granulocyte chemotactic protein-2 (GCP-2) is an important neutrophil chemotactic factor in the mouse that belongs to the CXC chemokine family. Although the local tissular effects of chemokines are well known, only recently has the systemic regulation of leukocytes become accepted. To study the pharmacokinetics of mouse GCP-2 and the systemic effects on leukocytes, we expressed a potent natural isoform of mouse GCP-2, GCP-2(9-78), in Escherichia coli and produced electrophoretically pure material. GCP-2(9-78) was 10-fold more potent to chemoattract neutrophils than recombinant GCP-2(5-78). After intravenous (i.v.) injection in mice, GCP-2(9-78) persisted in the circulation with an average half-life of 42 min. When a bolus of 1 mg/kg recombinant mouse GCP-2(9-78) was injected systemically, a significant effect on circulating leukocytes was observed. After a neutropenic phase, at its height at 1 h after injection, neutrophil numbers increased to a maximum at 4 h postinjection, and a concomitant decrease in lymphocyte numbers was observed. In control mice injected with isotonic saline, changes in leukocyte numbers were less pronounced and followed a different kinetic. Whereas tissular neutrophil chemotaxis to GCP-2 is influenced by gelatinase B, the systemic effects on neutrophilia and lymphopenia were not different in gelatinase B-deficient and wild-type mice. These data reinforce the idea that chemokines, including GCP-2, influence the homeostasis of circulating leukocyte numbers.
Starckx
Sofie
S
Rega Institute for Medical Research, Laboratory for Molecular Immunology, University of Leuven, Leuven, Belgium.
Wuyts
Anja
A
Opsomer
Inge
I
Van Coillie
Els
E
Proost
Paul
P
Arnold
Bernd
B
Van Damme
Jo
J
Opdenakker
Ghislain
G
D000818:Animals; D054427:Chemokine CXCL6; D019743:Chemokines, CXC / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology / Q000633:toxicity; D002634:Chemotaxis, Leukocyte / Q000187:drug effects* / Q000502:physiology; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism*; D006207:Half-Life; D006706:Homeostasis; D007275:Injections, Intravenous; D007958:Leukocyte Count; D007964:Leukocytosis / Q000139:chemically induced; D008231:Lymphopenia / Q000139:chemically induced; D020780:Matrix Metalloproteinase 9 / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009504:Neutrophils / Q000187:drug effects; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology / Q000633:toxicity; D011993:Recombinant Fusion Proteins / Q000493:pharmacokinetics / Q000494:pharmacology / Q000633:toxicity; D013047:Specific Pathogen-Free Organisms
D054427:Chemokine CXCL6; D019743:Chemokines, CXC; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D020780:Matrix Metalloproteinase 9