Enzyme Linked Immunosorbent Assay for Quantification of Bovine Interleukin‐8 to Study Infection and Immunity in the Female Genital Tract

Abstract

PROBLEM: The chemokine IL-8 recruits neutrophils to sites of infection, including the endometrium of the bovine uterus. However, quantification of bovine IL-8 often yields lower concentrations than for other species, which may reflect impaired innate immune responses by bovine cells or inaccurate measurement of IL-8 using the current human IL-8 ELISA method. METHOD OF STUDY: An ELISA was developed and validated for detection of bovine IL-8. Utility of the assay was tested by measuring the response of bovine endometrium and cells to bacteria and pathogen-associated molecular patterns. RESULTS: The developed ELISA detected 62.5-2000 pg/mL IL-8, with minimal cross-reactivity to other inflammatory mediators. Concentrations of bovine IL-8 were measured more accurately by the bovine than human IL-8 ELISA. Bovine endometrial IL-8 responses to pathogen-associated molecules were quantitatively similar to other species. CONCLUSION: A bovine-specific IL-8 ELISA was developed, which accurately measured IL-8 secretion from endometrial cells.