Isolation and characterization of an interleukin-8-like peptide in the bovine species.
Human interleukin-8 (IL-8) is a biologically active peptide which displays chemo-attractive activity for neutrophils and T-cells. The molecule is produced by a variety of cell types upon exposure to lipopolysaccharide, interleukin-1 and tumor necrosis factor. Recombinant human IL-8 also stimulates chemotaxis of bovine cells in a dose dependent manner. The purpose of this series of studies was to investigate the ability of bovine cells to produce an active IL-8-like molecule and to determine if bovine cells respond to human recombinant IL-8. Stimulation of purified peripheral blood mononuclear cells results in the time dependent production of an IL-8-like molecule as determined using an anti-human IL-8 ELISA assay and a bovine neutrophil chemotactic assay. Physical characterization indicates that the biological activity of the molecule was significantly reduced by heat inactivation at 56 degrees C for 30 min or exposure to extreme acidic or basic conditions. The peptide was affinity purified using an anti-human IL-8 antibody produced from ATCC hybridoma HB9647. SDS-PAGE analysis yields a distinct band at 7.8 kDa. The isoelectric point of the purified protein was determined to be 8.65. Biological activity of the purified protein was confirmed and the anti-human IL-8 antibody was capable of partially neutralizing the chemotactic activity.
Central Research Division, Pfizer, Inc., Terre Haute, IN 47808.
R L
RL
Hassfurther
P C
PC
Canning
R W
RW
Geib
D000818:Animals; D002417:Cattle / Q000276:immunology*; D002634:Chemotaxis, Leukocyte; D002846:Chromatography, Affinity / Q000662:veterinary; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D006801:Humans; D016209:Interleukin-8 / Q000096:biosynthesis / Q000302:isolation & purification*; D007526:Isoelectric Point; D009000:Monocytes / Q000276:immunology; D009504:Neutrophils / Q000276:immunology; D010455:Peptides; D011994:Recombinant Proteins
D016209:Interleukin-8; D010455:Peptides; D011994:Recombinant Proteins