Fig 1: LTC treatment of porcine PBMC stimulates the significant release of IFNα, IFNβ, NO, and ROS. Cultures of PBMC were seeded at 4.5 × 106 cells per well in 24-well plates and treated for 36 h with either nothing (untreated) or 1, 5, 10, or 25 µg/mL of LTC, or 300 ng/mL LPS. Supernatants were assayed by ELISA for the release of porcine IFNα (A) or IFNβ (B). Culture supernatants were assayed for NO release by the Griess reagent assay (C) and for the production of ROS by treatment with 10 µM H2DCFDA (D). Medium controls were included to rule out any detection of cytokines in the growth medium. Significant differences in Type I IFN, NO, and ROS were determined using a one-way ANOVA with p-values of **** ≤ 0.001, ** ≤ 0.01, and * ≤ 0.05.
Fig 2: Correlation analysis of NET markers and other markers detected in the BALF with the growth progression of A.pp in the BALF. The correlation analysis was performed with Spearman r for IAV-negative BALF (A, C, E, G, I, K, M, O, Q) and for IAV-positive BALF (B, D, F, H, J, L, N, P, R) and detected several significant positive correlations. The growth progression of A.pp correlates positively with cell-free DNA (A, B), sialic acid (M, N), and IFN-γ (Q, R) in the case of IAV-negative and IAV-positive BALF. The growth progression of A.pp correlates with the nucleosome (D) only in the case of IAV-positive BALF. The values for r, p, and n are given in each graph, and each square or triangle represents one BALF sample. Several markers in the BALF correlate positively with the growth progression of A.pp, but this was not significant. The dotted line (O–R) represents the detection limit of the ELISA for IFN-α (36 pg/ml) and IFN-γ (2 pg/ml), as all the samples where no signal was detected were set to the detection limit.
Fig 3: Different markers in the BALF of respiratory diseased pigs tested negative (IAV-negative) and positive (IAV-positive) for IAV (A–D) and the correlation to the IAV Ct value detected in BALF and/or lung tissue (E–H). The amounts of nicotinamide adenine dinucleotide (NAD) (A), sialic acid (B), IFN-α (C), and IFN-γ (D) were analyzed in the BALF of 21 IAV-positive, 24 IAV-negative, and five (four in NAD) healthy control animals. For each marker, a box and whisker plot with the minimum and maximum values and individual points is presented. A Mann–Whitney test was calculated (*p < 0.05). The correlation of the analyzed markers and the Ct value of IAV was performed with Spearman r resulting in no significant correlation, except for a significant negative correlation between IAV Ct value and IFN-α (G). A simple linear regression is depicted in the graph and the values for r, p, and n are given (E–H). The dotted line (C, D, G, H) represents the detection limit of the ELISA for IFN-α (36 pg/ml) and IFN-γ (2 pg/ml), as all the samples where no signal was detected were set to the detection limit.
Supplier Page from Thermo Fisher Scientific for Porcine IFN-alpha ELISA Kit