Identification and characterization of rhesus macaque interleukin-8.
To establish a direct link between IL-8 and inflammation in vivo, we first isolated the gene encoding rhesus macaque IL-8. The open reading frame directs the translation of a 101 amino acid (aa) precursor, which is 94% identical to human IL-8. Rhesus IL-8 was expressed in bacteria and purified to homogeneity with ion-exchange chromatography. Pure rhesus IL-8 was biologically active as measured by its ability to bind specifically to either rhesus (Kd = 0.5 nM) or human (Kd = 2 nM) IL-8 receptors and to promote in vitro chemotaxis of rhesus (EC50 = 2 nM) or human neutrophils (EC50 = 4 nM). Moreover, a mouse monoclonal antibody, DM/C7, which neutralizes human IL-8 activity, also recognized and neutralized (IC50 = 0.5-3.0 microgram/ml) rhesus IL-8 in vitro. Systemic administration of DM/C7 completely inhibited the dermal inflammation of rhesus ears induced by the external application of phorbol myristoyl acetate. These observations reveal that rhesus IL-8 is structurally and functionally similar to human IL-8 and suggests that IL-8 plays a prominent role in a primate model of inflammation.
Minnerly
J C
JC
Searle Research & Development, St. Louis, Missouri 63198, USA.
Baganoff
M P
MP
Deppeler
C L
CL
Keller
B T
BT
Rapp
S R
SR
Widomski
D L
DL
Fretland
D J
DJ
Bolanowski
M A
MA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004487:Edema / Q000139:chemically induced / Q000503:physiopathology / Q000517:prevention & control; D005260:Female; D005796:Genes; D006801:Humans; D007249:Inflammation; D016209:Interleukin-8 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D008253:Macaca mulatta / Q000235:genetics / Q000378:metabolism*; D008322:Mammals / Q000235:genetics; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000187:drug effects; D016366:Open Reading Frames; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013755:Tetradecanoylphorbol Acetate / Q000633:toxicity
D000911:Antibodies, Monoclonal; D016209:Interleukin-8; D013755:Tetradecanoylphorbol Acetate