Development and Application of a Pig IL-8 ELISA Detection System

Abstract

Interleukin 8 (IL-8) is a chemotactic and activating chemokine, especially for neutrophils, which plays an important role in inflammatory process. A pig IL-8 specific enzyme-linked immunosorbent assay (ELISA) was developed to measure IL-8 concentrations in cell culture supernatants and biological fluids. A streptavidin-biotin amplified sandwich method uses mouse capture mAb IZ8.03 and detection biotinylated mouse mAb IZ8.04 against recombinant pig IL-8. The assay specifically and reproducibly recognizes both recombinant and natural pig IL-8. A working range of the assay is 16-1000 pg/mL and takes a mere 3.5 h of incubation time. This pig IL-8 ELISA is a suitable alternative way of measurement of IL-8 concentrations to time consuming and laborious IL-8 bioassays.

Impact of the Lipopolysaccharide Chemotype of Salmonella Enterica Serovar Typhimurium on Virulence in Gnotobiotic Piglets

“…IL‑8 was detected as described elsewhere [55], while TNF-α and IL-10 were measured by commercial kits (Life Technologies, Carlsbad, CA, USA). IL-8, TNF-α, and IL-10 were measured with the sensitivities 20 pg/mL, 15 pg/mL, and 15 pg/mL, respectively.…”

Interference of Bifidobacterium choerinum or Escherichia coli Nissle 1917 with Salmonella Typhimurium in gnotobiotic piglets correlates with cytokine patterns in blood and intestine

“…All samples with added protease inhibitor cocktail (Roche) were frozen immediately and kept at −70°C until used. The sandwich IL‐8 ELISA with a sensitivity of 15 pg/ml is described elsewhere [32]. IL‐10 and TNF‐α were detected with the same sensitivity of 15 pg/ml using a swine IL‐10 CytoSet™ and swine TNF‐α CytoSet™ (Invitrogen, Carlsbad, CA, USA), according to the manufacturer's instructions.…”

Immune Response of Gnotobiotic Piglets against Mycoplasma hyopneumoniae

“…IFN-γ, IL-4, and IL-10 levels were also measured by commercial ELISA kits (Swine Cytoset kit, Biosource International Inc., Camarillo, CA, U.S.A.). The concentration of porcine IL-8 was determined as described previously [24]. The concentration of porcine IL-18 was measured as described previously [17].…”

“…These samples were stored at -20°C until use. The concentration of IL-1β, IL-4, IL-8, IL-10, IL-18, TNF-α, and IFN-γ in the BALF was measured as described above [17,24].…”