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Enzyme Linked Immunosorbent Assay for Quantification of Bovine Interleukin‐8 to Study Infection and Immunity in the Female Genital Tract
Author(s) -
Cronin James G.,
Hodges Rebecca,
Pedersen Sara,
Sheldon Iain M.
Publication year - 2015
Publication title -
american journal of reproductive immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.071
H-Index - 97
eISSN - 1600-0897
pISSN - 1046-7408
DOI - 10.1111/aji.12344
Subject(s) - pathogen , biology , immune system , immunology , chemokine , immunity , innate immune system , interleukin , microbiology and biotechnology , parasite hosting , endometrium , cytokine , endocrinology , world wide web , computer science
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.