Journal: Microorganisms

Article Title: Polyethylene Terephthalate Hydrolases in Human Gut Microbiota and Their Implications for Human Health

doi: 10.3390/microorganisms12010138

Figure Lengend Snippet: Effects of PET MPs and hydrolysates on cell viability and macrophage polarization. ( A , B ) Relative expression levels of IL-1β, IL-6, TNF-α, and Arg-1 (IL-4 stimulation) in mouse monocyte macrophages RAW 264.7 cells treated with TPA (0.01 mM) ( A ) and TPA (1.00 mM) ( B ) for 24 h followed by LPS/IL-4 stimulation for 6 h. ( C ) Relative expression levels of IL-1β, IL-6, TNF-α, and Arg-1 (IL-4 stimulation) in mouse bone-marrow-derived macrophages (BMDMs) treated with TPA (1.00 mM) for 24 h followed by LPS/IL-4 stimulation for 6 h. ( D , E ) Relative expression levels of IL-1β ( D ) and Arg-1 ( E ) in RAW 264.7 cells treated with PET MPs (<400 μm) at a gradient dose at 0.01, 0.10, 0.50, and 1.00 mg/mL for 24 h followed by LPS/IL-4 stimulation for 6 h. ( F , G ) Relative expression levels of IL-1β ( F ) and Arg-1 ( G ) in RAW 264.7 cells treated with PET MPs (<400 μm) at a gradient dose at 0.01, 0.10, 0.50, and 1.00 mg/mL for 24 h without any stimulation. *** p -value < 0.001; ** p -value < 0.01; * p -value < 0.05.

Article Snippet: Macrophage colony-stimulating factor (M-CSF) (product code: HY-P7085), lipopolysaccharides (LPS) (product code: HY-D1056), and murine interleukin-4 (IL-4) (product code: HY-P70644) were obtained from MedChemExpress, Princeton, NJ, USA.

Techniques: Expressing, Derivative Assay

Journal:

Article Title: Production of Borreliacidal Antibody to Outer Surface Protein A In Vitro and Modulation by Interleukin-4

doi:

Figure Lengend Snippet: Borreliacidal antibody response of lymph node cells obtained from 17-day-vaccinated mice cultured with macrophages and B. burgdorferi and treated with anti-murine IL-4 10 min (checkered bar) or 4 days (cross-hatched bar) after cultivation. Control cultures (solid bar) were treated with a non-isotype-specific antibody. Titers of borreliacidal antibody did not vary by more than one dilution in replicate assays. In most cases (90%) the same titer was obtained.

Article Snippet: In other experiments, recombinant IL-4 (rIL-4) or rat anti-murine IL-4 (R&D Systems, Minneapolis, Minn.) at quantities ranging from 0.01 to 1.0 μg was added to cultures of immune lymph node cells, macrophage, and B. burgdorferi at 10 min and 4 days of incubation.

Techniques: Cell Culture

Journal:

Article Title: Type 1 Immunity Provides Optimal Protection against Both Mucosal and Systemic Trypanosoma cruzi Challenges

doi: 10.1128/IAI.70.12.6715-6725.2002

Figure Lengend Snippet: RNase protection assays demonstrate that the type 1 and type 2 biased intranasal vaccination protocols induced highly polarized immune responses. Total spleen cells (4 × 106 cells/ml) were harvested and rested in medium alone, stimulated with T. cruzi lysate, or stimulated with ConA, using methods similar to those described in the legend to Fig. ​Fig.1.1. Total RNA samples were harvested from parallel cultures 24 and 48 h later using RNeasy Kits (Qiagen). These RNA samples were used in RiboQuant RNase protection assays (Pharmingen) to assess the levels of cytokine mRNA expressed. The autoradiogram shown is representative of multiple similar experiments. A progressive increase in IFN-γ mRNA expression induced by T. cruzi lysate in vitro was the most prominent finding with spleen cells from type 1 biased mice. On the other hand, parasite-specific increases in IL-4, IL-5, IL-9, and IL-13 mRNA were detected with spleen cells from type 2 biased mice, especially after 48 h of in vitro stimulation.

Article Snippet: Mice given the type 2 bias immunization protocol received 2,500 U of recombinant murine IL-4 (Pharmingen, San Diego, Calif.) and 0.5 mg of the anti-IFN-γ monoclonal antibody R46A2 instead of IL-12 and 11B11.

Techniques: Expressing, In Vitro

Journal:

Article Title: Type 1 Immunity Provides Optimal Protection against Both Mucosal and Systemic Trypanosoma cruzi Challenges

doi: 10.1128/IAI.70.12.6715-6725.2002

Figure Lengend Snippet: Cytokine secretion studies confirm that type 1 and type 2 biased immunization protocols induce highly polarized T. cruzi-specific immunity. BALB/c mice were immunized as described in Table ​Table1.1. Three days after the final intranasal antigen challenge, mice were sacrificed and spleen cell suspensions were prepared. (A) Total spleen cells (2 × 105 cells/200 μl) were stimulated with T. cruzi lysate, and 72 h later the culture supernatants were harvested and assayed for secreted IFN-γ and IL-4 levels by ELISA. (B) Similar studies were done with purified CD4+ T cells (5 × 104 cells/200 μl) cultured with irradiated syngeneic total spleen cells (106 cells/200 μl) pulsed with T. cruzi lysate. Both sets of data had background levels of cytokines spontaneously produced in medium-rested cultures subtracted from the values presented. Shown are the means and standard errors from a single experiment with pooled cells derived from two to three mice/group. These results are representative of at least five similar experiments. Similar results were seen in lymphocytes harvested from Peyer's patches and from mesenteric lymph nodes (data not shown).

Article Snippet: Mice given the type 2 bias immunization protocol received 2,500 U of recombinant murine IL-4 (Pharmingen, San Diego, Calif.) and 0.5 mg of the anti-IFN-γ monoclonal antibody R46A2 instead of IL-12 and 11B11.

Techniques: Enzyme-linked Immunosorbent Assay, Purification, Cell Culture, Irradiation, Produced, Derivative Assay