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Characterization of the Bovine Innate Immune Response to Intramammary Infection with Klebsiella pneumoniae

¹ Bovine Functional Genomics Laboratory and
² Veterinary Services, USDA-Agricultural Research Service, Beltsville, MD 20705
³ Institute for Animal Health, Compton, Near Newbury, Berkshire RG20 7NN, UK

Corresponding author: D. D. Bannerman; e-mail: dbanner{at}anri.barc.usda.gov.

Gram-negative bacteria are responsible for almost one-half of the clinical cases of mastitis that occur annually. Of those gram-negative bacteria that induce mastitis, Klebsiella pneumoniae remains one of the most prevalent. Detection of infectious pathogens and the induction of a proinflammatory response are critical components of host innate immunity. The objective of the current study was to characterize several elements of the bovine innate immune response to intramammary infection with Klebsiella pneumoniae. The inflammatory cytokine response and changes in the levels of soluble CD14 (sCD14) and lipopolysaccharide (LPS)-binding protein (LBP), 2 proteins that contribute to host recognition of gram-negative bacteria, were studied. The contralateral quarters of 7 late-lactating Holstein cows were challenged with either saline or K. pneumoniae, and milk and blood samples were collected. Initial increases in the chemoattractants C5a and IL-8, as well as TNF-, were evident in infected quarters within 16 h of challenge and were temporally coincident with increases in milk somatic cells. Augmented levels of TNF- and IL-8 were observed in infected quarters until >48 h postchallenge, respectively. Elevated levels of IL-12, IFN-, and the antiinflammatory cytokine, IL-10, which were first detected between 12 and 20 h postinfection, persisted in infected quarters throughout the study (>96 h). Initial increases in milk LBP and sCD14 were detected 16 and 20 h, respectively, after challenge. Together, these data demonstrate that intramammary infection with K. pneumoniae elicits a host response characterized by the induction of proinflammatory cytokines and elevation of accessory molecules involved in LPS recognition.

Key Words: lipopolysaccharide-binding protein • Klebsiella pneumoniae • mastitis • innate immunity

Abbreviation key: HRP = horseradish peroxidase, LBP = LPS-binding protein, mCD14 = membrane-bound CD14, OD = optical density, sCD14 = soluble CD14, TBS = Tris-buffered saline, TMB = tetramethylbenzidine, Tlr = toll-like receptor

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