Production and characterization of polyclonal anti-canine interleukin-8 antibodies.
Polyclonal anti-canine interleukin-8 (cIL-8) antibodies were raised in rabbits immunized with cIL-8 expressed by E. coli. Polyclonal antibodies were purified by affinity chromatography. In the enzyme linked immunosorbent assay (ELISA), the resulting anti- cIL-8 antibodies showed relatively high reactivities with cIL-8 in the fusion proteins of glutathione-S-transferase/cIL-8 (GST/cIL-8) and maltose binding protein/cIL-8 (MBP/cIL-8), but negligible ones with MBP. Furthermore, Western blotting analysis using these polyclonal antibodies showed distinct bands for cIL-8, GST/cIL-8, and MBP/cIL-8. These antibodies also bound to recombinant human IL-8 (rhIL-8) in ELISA but not in Western immunoblotting. The rHIL-8 (50-800 ng/ml) was chemoattractant for canine neutrophils in a dose dependent manner, but the anti-cIL-8 antibodies did not show the inhibitory effect on the chemotactic activity of rhIL-8 of canine neutrophils, when tested by the chemotaxis assay using Boyden chambers. In addition, GST/cIL-8 and rhIL-8 induced strong and rapid shape change responses of canine neutrophils. However, the anti-cIL-8 antibodies inhibited shape change responses induced by GST/cIL-8 but not by rhIL-8.
Mohamed
A
A
Department of Molecular Immunology, Faculty of Agriculture, University of Tokyo, Japan.
Matsumoto
Y
Y
Furusawa
S
S
Yoshihara
K
K
Matsumoto
Y
Y
Suzuki
K
K
Onodera
T
T
Hirota
Y
Y
D018528:ATP-Binding Cassette Transporters; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000917:Antibody Formation; D000918:Antibody Specificity; D015153:Blotting, Western; D002352:Carrier Proteins; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D006801:Humans; D016209:Interleukin-8 / Q000032:analysis* / Q000276:immunology*; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology
D018528:ATP-Binding Cassette Transporters; D000906:Antibodies; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D016209:Interleukin-8; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C461599:maltose transport system, E coli