Table 1.
Primary antibodies used in study.
Table 2.
Secondary antibodies used in studya.
Fig 1.
Schematic outlining relationships and properties of polarized macrophages.
Fig 2.
Mean number (Log μm2) of macrophages (AM-3K) classified as host defense (IFN-γ, IL-1β, IL-12, IL-23, uNOS, TNF-α, and TLR4) or regulation of inflammation (IL-1Ra, IL-10m CD163, CD206, and TGF-β), present in frozen bovine mid-ileal tissue sections collected from cows naturally infected with Mycobacterium avium subsp. paratuberculosis (subclinical and clinical), compared to uninfected control cows.
Asterisks denote statistical significance: * p<0.05; ** p<0.01; *** p<0.001; **** p<0.0001.
Fig 3.
Representative IF labeling in frozen bovine mid-ileal tissue, comparing cows subclinically and clinically infected with Mycobacterium avium subsp. paratuberculosis (MAP).
Nuclear staining was performed using 4’,6- diamidino-2-phenylindole, dihydrochloride (DAPI, blue). Each merged image was selected as an example of representative IF labeling from a single cow. (A and E) CD163; (B and F) TLR4; (C and G) AM-3K; (D and H) merged image of CD163, TLR4 and AM-3K; (I and M) CD206; (J and N) uNOS; (K and O) AM-3K; (L and P) merged image of CD206, uNOS and AM-3K.
Fig 4.
Mean number (Log μm2) of total macrophages (AM-3K) classified as either host defense or regulation of inflammation macrophages.
IF staining was performed on frozen bovine mid-ileal tissue sections collected from cows naturally infected with Mycobacterium avium subsp. paratuberculosis (subclinical and clinical), compared to uninfected control cows. Asterisks denote statistical significance: ** p<0.01.
Fig 5.
Pairwise Pearson correlations for total number of macrophages (Mφ) with host defense Mφ phenotypes.
(A) correlation between total Mφ and number of IFN-γ+ Mφ for control cows; (B) correlation between total Mφ and number of IL-12+ Mφ for control cows (○ and solid line) and subclinical cows (● and dotted line); (C) correlation between total Mφ and number of IL-1β+ Mφ for subclinical cows; and (D) correlation between total Mφ and number of IL-23+ Mφ for subclinical cows).
Table 3.
Pairwise Pearson correlations between cytokines colocalized within host defense macrophages.
Fig 6.
Pairwise Pearson correlations for total number of macrophages (Mφ) with resolution and repair Mφ phenotypes.
(A) correlation between total Mφ and number of IL-1Ra+ Mφ for subclinical cows; (B) correlation between total Mφ and number of IL-10+ Mφ for subclinical cows (● and dotted line) and clinical cows (□ and dotted line); (C) correlation between total Mφ and number of TGF-β+ Mφ for subclinical cows; and (D) correlation between total Mφ and number of CD206+ Mφ for subclinical cows.
Table 4.
Pair-wise Pearson correlations between cytokines colocalized within resolution and repair macrophage phenotypes.