Table 1.
Demographic data.
Fig 1.
Representative flow cytometry gating strategy.
Alveolar cells were surface stained for CD45, CD4, CCR6 and then intracellularly stained for IL-17A and Interferon-gamma (IFN-ɣ). We gated on single cells, CD45+ cells, live CD4+ T cells, and then for either CCR6+IL17A+ cells or total IFN-ɣ producing cells. SSH: Side Scatter Height, SSA: Side Scatter Area.
Fig 2.
Percentage of Th17 cells reduced in the setting of ventilator associated pneumonia.
Cells were isolated from bronchoalveolar lavage fluid, stained, and percentages of Th17 cells and CD4+ IFN-γ+ positive cells were obtained using flow cytometry. A: Percentage of Th17 cells (CCR6+IL-17A+) of the total CD4+ population in the BAL of patients with VAP or without VAP. B. Percentage of IFN-γ+ cells of the total CD4+ population. C. IL-17A protein concentrations measured by immunoassay in the BAL of subjects with and without VAP. D. We compared the relationship between numbers of Th17 cells (x-axis) and amount of IL-17A protein (y-axis) in subjects with VAP (triangles) and without VAP (circles).
Fig 3.
Increased levels of pro-inflammatory cytokines in bronchoalveolar lavage samples of patients with ventilator associated pneumonia.
Bronchoalveolar lavage fluid was isolated from patients with and without ventilator associated pneumonia (VAP) and run on a multi-plex immuno assay to measure cytokine protein levels. Subjects with VAP (red triangles) are compared to those without VAP (black circles). All y-values are the log of the concentration in pg/mL. p-values were measured using a 2-tailed T-test. For most pro-inflammatory cytokines, such as A: TNF-α, B: IFN-γ, C: IL-1β, and E: IL-12p 70, and for the chemokine IL-8 (D), there was increased expression in the VAP samples. For IL-6 (F) and IL-10 (G), there was no significant difference between the two groups.
Fig 4.
Increased protein in bronchoalveolar lavage correlates to levels of IL-17A protein.
Total protein level was measured in BAL using an ELISA. A. There is no significant difference in the amount of protein between the VAP and no VAP groups (p = 0.2). B. Correlation between log of IL-17A cytokine concentration (x-axis) and log of protein concentration (y = axis). C. Correlation between percentage of Th17 cells (x-axis) and log of protein concentration (y-axis).
Fig 5.
Neutrophil numbers in BAL correlate to IL-17A protein levels.
Cytospins were counted for total numbers of neutrophils present in each BAL sample. Correlation between percentage of neutrophils (x-axis) and A) IL-17A protein concentration (log, y-axis) and B) percentage of Th17 cell numbers.