Fig 1: Neutrophil recruitment and proinflammatory cytokine production after challenge determined by flow cytometry and multiplex immunoassay analysis. (A) Blood neutrophil count of Sprague-Dawley rats infected with Bordetella pertussis. At days 1, 3, 6, 9, and 12 postinfection, rats were euthanized, and blood was collected via cardiac puncture. Flow cytometry was used to quantify the amount of neutrophils (CD45+ CD161− B220– CD43+ His48hi) in the blood. Quantification of the percentage of single, CD45+ cells was performed. The dotted line represents the average frequency of CD45+ neutrophils in the mock challenge animals over the course of the study. (B and C) Analysis of cytokine IL-6 from the supernatant of lung homogenate and serum. (D and E) Analysis of cytokine IL-17 in serum and lung homogenate supernatant. Cytokines were analyzed using a ProcartaPlex multiplex immunoassay kit. Results are shown as means ± the SEM (n = 4). The dotted line represents the average mock-challenged cytokine levels measured. P values were determined by one-way ANOVA, followed by a Tukey comparison test (#, P < 0.05; ##, P < 0.01; ####, P < 0.0001 [compared to mock challenge]).