Production and Characterization of Polyclonal Anti-Canine Interleukin-8 Antibodies

[摘要] References(25)Cited-By(2)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 for 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.

[学科分类] 兽医学

[关键词] anti-canine interleukin-8 antibody;chemotaxis;interleukin-8