Complement fragment C5a and inflammatory cytokines in neutrophil recruitment during intramammary infection with Escherichia coli

Abstract

Generation of inflammatory mediators and leukocyte recruitment to infection at an epithelial surface were studied during Escherichia coli-induced mastitis. One uninfected gland of each of eight midlactation cows was challenged with only 30 CFU of E. coli McDonald strain 487, a serum-resistant isolate from a cow with mastitis. Bacteria grew logarithmically during the first 10 to 12 h after challenge, reaching concentrations of more than 10 5 CFU/ml with no detectable host response during this time. An intense inflammatory reaction began approximately 12 h after the challenge and was characterized by a breakdown in the blood-milk permeability barrier followed by pyrexia and a pronounced leukocytic influx. Coincident with the onset of mammary inflammation was the appearance of neutrophil chemotactic activity in the milk from infected glands. Factors able to upregulate CD18 expression on peripheral blood neutrophils also appeared in milk at this time. The lack of appearance of chemotactic and CD18-upregulating activities until 12 h after challenge indicated that delays in neutrophil recruitment resulted from an initial lack of bacterial recognition and inflammatory mediator production. Production of complement fragment C5a, tumor necrosis factor, and interleukin-1 (IL-1) occurred earlier than production of IL-6 or IL-8. The early and intense production of C5a indicates that this chemoattractant may be more important than IL-8 during the initial recruitment and activation of neutrophils to a developing E. coli infection.

Modulatory Effects of Bovine Seminal Plasma on Uterine Inflammatory Processes

“…2000), whereas Shuster et al. observed cytokine and complement secretion following an Escherichia coli ‐induced mastitis (Shuster et al. 1997).…”

“…For the analysis of IL‐8 concentrations in cell culture supernatants with and without PMN, a commercial ELISA‐kit (Quantikine ® , IL‐8 ELISA; R&D Systems, Wiesbaden, Germany) approved in detecting bovine IL‐8 (Shuster et al. 1997; Boudjellab et al.…”

The Cytokine Markers inStaphylococcus aureusMastitis of Bovine Mammary Gland

“…Although S. aureus and coliform mastitis are associated with high influx of neutrophils, their activation state and CD markers expression are different. Shuster et al (1997) showed that recruited neutrophils expressed strong chemotactic activity with the ability to upregulate CD18. Whereas, neutrophils form quarters chronically infected with S. aureus were less active and with low tendency to express CD18 and CD11b (Riollet et al, 2001).…”

Increased Levels of LPS-Binding Protein in Bovine Blood and Milk Following Bacterial Lipopolysaccharide Challenge

“…Milk IL-8 levels were determined from undiluted whey samples assayed with a commercially available human IL-8 ELISA kit (R&D Systems, Inc., Minneapolis, MN). The antibody pairs used in this kit have been previously shown to cross-react with bovine IL-8 (Shuster et al, 1996(Shuster et al, , 1997. The optical density at 450 nm and a correction wavelength of 550 nm were measured on a microplate reader.…”