Table 1.
Primers used for qRT-PCR.
Table 2.
Absolute cytokine level after LPS induction.
Fig 1.
Comparison of serum chemistry and histopathology.
(A) Experimental schematic: For chronic inflammation, 5 μg/kg lipopolysaccharide (LPS) was administrated, intramuscularly (IM), seven times, daily. For acute inflammation, 25 μg/kg LPS was administrated, IM, once at day7. Two CMS minipigs were used for each group (control, chronic, acute). Blood and tissue were sampled for further study. (B) Serum chemistry was measured using a TBA 120 FR chemistry analyzer (Toshiba Co., Japan). Absolute values are indicated. #P<0.05. (C) H&E staining. Glomeruli in kidney and central vein in liver are indicated with a black dot line. Abnormal lesions are indicated with black arrow head. Scale bar = 100 μm.
Fig 2.
Immune cell distribution in spleen.
(A–D) Immunohistochemistry. Population and distribution of (A) CD11b+ (macrophage), (B) MPO+ (neutrophil), (C) CD4+, and (D) CD8+ (lymphocyte) cells were examined in spleen. Representative images are shown; N≥3. Quantification of the immune cell number is shown as mean ± SD; #P<0.01 relative to control, *P<0.05 relative to control. Scale bar = 20 μm.
Fig 3.
Immune cell distribution in lung.
(A–D) Immunohistochemistry. Population and distribution of (A) CD11b+ (macrophage), (B) MPO+ (neutrophil), (C) CD4+, and (D) CD8+ (lymphocyte) cells were examined in lung. Representative images are shown; N≥3. Quantification of the immune cell number is shown as mean ± SD; aP<0.01 relative to trachea control, bP<0.05 relative to trachea control, cP<0.01 relative to alveoli control, dP<0.05 relative to alveoli control. Scale bar = 20 μm.
Fig 4.
Inflammation-related gene expression in tissues and PBMC.
(A) qRT-PCR and Heatmap in tissues. IL-6, CRP, COX-2, SOD1 mRNA expression were analyzed in heart, lung, liver, kidney, and duodenum under chronic and acute inflammation with two biological and three experimental replicates. Red color indicates gene upregulation and green color indicates gene downregulation. (B) qRT-PCR in PBMC. Pro-inflammatory (IL-1β, IL-6, TNFα, COX-2) and anti-inflammatory (TGFβ, IL-10) mRNA expression were analyzed in PBMC under chronic and acute inflammation with two biological and three experimental replicates.
Fig 5.
Serum cytokine analysis using ELISA.
Blood was collected at day 0, 1, 2, 3, 4, 5, 6, 7, and 8 and then serum was isolated. Absolute cytokine levels (pg/ml) were measured by ELISA with two biological and three experimental replicates. (A) IL-1β, (B) IL-6, (C) TNFα, (D) IL-8, (E) IFNγ. U.D. = undetected.