Journal: Journal of Virology

Article Title: Pathobiology of an NS1-Truncated H3N2 Swine Influenza Virus Strain in Pigs

doi: 10.1128/jvi.00519-22

Figure Lengend Snippet: wtTX98 and lvTX98 replication and IFN-α/β-inducing capacity in PK-15 cells. PK-15 cell monolayers were infected with a MOI of 0.001 or a MOI of 2 of wtTX98 (blue circles) or lvTX98 (orange squares), and supernatant samples were evaluated for virus titers in a CPE assay on MDCK cells (A) and for bioactive IFN-α/β titers in a CPE reduction assay with VSV on MDBK cells (B) at different time points postinoculation. Means with standard deviations of log 10 -transformed virus titers, log 2 -transformed IFN-α/β titers, and areas under the curve of 5 replicates are shown. Black dotted lines represent the detection limit. Results were compared between groups using 2-sided Mann-Whitney U tests. *, P < 0.05.

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) kits used for detection of IFN-α and IFN-β in 20-times-concentrated BAL fluids were the Verikine-HS pig interferon alpha ELISA kit from PBL Assay Science (catalog no. 47100-1, lot no. 7031, detection range of 2.34 to 150 pg/mL) and the porcine IFN-β ELISA kit from Emelca Bioscience (catalog no. MBS760799-2, lot no. P0074E040, detection range of 15.625 to 1,000 pg/mL).

Techniques: Infection, Transformation Assay, MANN-WHITNEY

Journal: Journal of Virology

Article Title: Pathobiology of an NS1-Truncated H3N2 Swine Influenza Virus Strain in Pigs

doi: 10.1128/jvi.00519-22

Figure Lengend Snippet: Proinflammatory cytokines in BAL fluid of pigs intratracheally inoculated with wtTX98 or lvTX98. Conventional 6-week-old influenza-naive pigs were intratracheally inoculated with 7.5 log 10 TCID 50 of wtTX98 (●, n = 15) or lvTX98 (■, n = 15) in 3 mL of PBS. Three pigs per group were euthanized at different time points, and BAL fluid was collected. After removal of BAL cells, cell-free BAL fluid was concentrated 20 times for cytokine analysis. (A) Bioactive IFN-α/β titers as determined in a CPE reduction assay with VSV on MDBK cells. (B) IFN-α ELISA titers. (C) IFN-β ELISA titers. (D) Bioactive IL-6 titers as determined in a proliferation assay with B9 cells. Dots represent log 2 -transformed cytokine titers for individual pigs; blue and orange bars represent mean values for wtTX98- and lvTX98-inoculated pigs, respectively, at each time point. Black dotted lines represent the detection limit. Results were compared between groups using 2-sided Mann-Whitney U tests. # , P < 0.1; #, P = 0.1.

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) kits used for detection of IFN-α and IFN-β in 20-times-concentrated BAL fluids were the Verikine-HS pig interferon alpha ELISA kit from PBL Assay Science (catalog no. 47100-1, lot no. 7031, detection range of 2.34 to 150 pg/mL) and the porcine IFN-β ELISA kit from Emelca Bioscience (catalog no. MBS760799-2, lot no. P0074E040, detection range of 15.625 to 1,000 pg/mL).

Techniques: Enzyme-linked Immunosorbent Assay, Proliferation Assay, Transformation Assay, MANN-WHITNEY

Journal: Frontiers in Veterinary Science

Article Title: Expression Dynamics of Innate Immunity in Influenza Virus-Infected Swine

doi: 10.3389/fvets.2017.00048

Figure Lengend Snippet: (A) Interferon (IFN)-α was investigated in bronchoalveolar fluids (BALF) samples of Swine IV-infected pigs at day 3 p.i. by sandwich ELISA with monoclonal antibodies F17 and K9 to porcine IFN-α1. Results are shown in terms of Units/ml (mean + 1 SD) on the basis of a standard curve created with porcine recombinant IFN-α1. IFN-α titers <1 U/ml were the baseline in our assays. The BALF samples of mock and all other IV groups were always IFN α-negative at all the time points. (B) Interleukin (IL)-6 was measured in BALF samples by a bioassay on IL 6-dependent 7TD1 hybridoma cells. On the basis of both final number and viability of 7TD1 cells, IL-6 concentration was determined from a standard curve created with a reference preparation of human recombinant IL-6 (Pierce Endogen, Rockford, IL, USA). Results are shown in terms of group average +1 SEM. The IL-6 concentration in the swine group at day 3 p.i. is significantly different (one-way ANOVA, P < 0.05). Please notice that the result of the Seal group at day 3 p.i. was accounted for by an outlier and three non-responder pigs. Samples from day 3 p.i. are shown as gray bars, from day 6 p.i. as striped bars, and from day 21 p.i. as black bars.

Article Snippet: Identification of the cytokine was performed by a neutralization assay on MDBK cells of IFN α-positive BALF samples , using monoclonal antibody (mAb) G16 to porcine IFN-α1 (Serotec, cat. MCA1935Z).

Techniques: Infection, Sandwich ELISA, Bioprocessing, Recombinant, Bioassay, Concentration Assay

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Decreased PPAR-γ expression after internal carotid artery stenting is associated with vascular lesions induced by smooth muscle cell proliferation and systemic inflammation in a minipig model

doi:

Figure Lengend Snippet: Immunohistochemical staining for peroxisome proliferator-activated receptor gamma (PPAR-γ) expression in the vascular smooth muscle cells (VSMCs) in each group. The immunohistochemical staining was conducted 90 days after stenting. A: Normal control group, B: Stenting group, C: Stenting plus rosiglitazone (ROSI) treatment group. PPAR-γ-positive VSMCs are shown by their nuclear brown staining (×100).

Article Snippet: Immunohistochemistry and western blotting After deparaffinization and hydration, the specimens were blocked with 0.3% H 2 O 2 at room temperature for 15 min. A rabbit polyclonal antibody against pig-PPAR-γ (2.5 μg/ml, ABD Serotec, USA) was used for immunohistochemical staining.

Techniques: Immunohistochemical staining, Staining, Expressing, Control

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Decreased PPAR-γ expression after internal carotid artery stenting is associated with vascular lesions induced by smooth muscle cell proliferation and systemic inflammation in a minipig model

doi:

Figure Lengend Snippet: Western blot assay for the expression of minipig carotid artery peroxisome proliferator-activated receptor gamma (PPAR-γ) and smooth muscle 22-alpha (SM22α). The carotid artery tissues were harvested and the total protein was extracted for the western blot analysis by using anti-PPAR-γ, anti-SM22α, and anti-β-actin primary antibodies. The immunocomplexes were detected using horseradish peroxidase-conjugated secondary antibodies, followed by enhanced chemiluminescence. The relative molecular masses of the target proteins are indicated.

Article Snippet: Immunohistochemistry and western blotting After deparaffinization and hydration, the specimens were blocked with 0.3% H 2 O 2 at room temperature for 15 min. A rabbit polyclonal antibody against pig-PPAR-γ (2.5 μg/ml, ABD Serotec, USA) was used for immunohistochemical staining.

Techniques: Western Blot, Expressing