Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Type I IFN-producing CD4 Valpha14i NKT cells facilitate priming of IL-10-producing CD8 T cells by hepatocytes.

doi: 10.4049/jimmunol.178.4.2083

Figure Lengend Snippet: FIGURE 6. The cytokine response of OT-I CD8 T cells primed by HC with or without NKT cell help. A, OT-I CD8 T cells (105/well) were cocul- tured with GalCer- and SIINFEKL peptide-pulsed HC (104/well) with () or without () NKT cells (4 104/well). Cells were harvested after a 72-h culture, incubated for 4 h with ionomycin (750 ng/ml) and PMA (50 ng/ml), washed, and tested for IL-10 secretion in a 3-h capture assay. Cells were labeled with a PE-conjugated anti-IL-10 mAb and an anti-CD8 mAb. Data from a representative of five independent experiments are shown. B, OT-I CD8 T cells (105/well) were cocultured for 72 h with GalCer-/ SIINFEKL peptide-pulsed HC (104/well) with () or without () NKT cells (4 104/well). CD8 T blasts were harvested, purified, and restimu- lated in vitro for 48 h with peptide-pulsed DC. IFN-, IL-2, and IL-10 in 48-h supernatants of these secondary cultures were determined by ELISA. Mean values of triplicates (SEM) of a representative of four independent experiments are shown.

Article Snippet: IL-10-producing CD8 T cells were identified by a cytokine capture assay (catalog no. 130-090-489; Miltenyi Biotec) combined with surface staining with the allophycocyanin-conjugated anti-CD8 mAb 53-6.7 (catalog no. 553035; BD Biosciences).

Techniques: Incubation, Labeling, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: Life Science Alliance

Article Title: Gene-selective transcription promotes the inhibition of tissue reparative macrophages by TNF

doi: 10.26508/lsa.202101315

Figure Lengend Snippet: (A, B, C) RNA from wild-type BMDMs left untreated (Ctrl), stimulated with IL4/IL13, IL4/IL13 + TNF (A), IL4/IL13 + IL1b (B) or IL4/IL13 + IL36 (C) was used for qRT-PCR. Data shown are the mean fold-increase compared with the Ctrl group. (D) TnfR1 −/− BMDMs were stimulated with IL4/IL13 or IL4/IL13 + TNF over time. RNA was isolated and analyzed for Retnla expression. Data shown are the mean fold-increase of the Ctrl group. All values are means ± SEM; **** P < 0.0001. Statistically significant differences were determined by one-way ANOVA with Tukey correction (A, B, C) or by two-way ANOVA (D). If not indicated otherwise superscripts show statistical significance compared with the control group. n = 3 biological replicates.

Article Snippet: Where indicated, BMDMs were stimulated with 10 ng/ml IL4 (produced in insect cells), 10 ng/ml IL13 (210-13; Peprotech), 10 ng/ml TNF (315-01A; Peprotech), 10 ng/ml IL1β (401-ML/CF; R&D System), 33.3 ng/ml IL36α (7059-ML/CF; R&D Systems), 33.3 ng/ml IL36β (7060-ML/CF; R&D Systems), 33.3 ng/ml IL36γ (6996-IL/CF; R&D Systems), 5 ng/ml LPS from Escherichia coli O111:B4 (L4391; Sigma-Aldrich), 10 μg/ml Etanercept (Erelzi; Sandoz), 20 μM JNK-IN-8 (SML1246; Sigma-Aldrich), and 10 μM SP600126 (BML-EI305; Enzo).

Techniques: Quantitative RT-PCR, Isolation, Expressing, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Hydrangea serrata extract attenuates PM-exacerbated airway inflammation in the CARAS model by modulating the IL-33/ST2/NF-κB signaling pathway.

doi: 10.1016/j.biopha.2024.116596

Figure Lengend Snippet: Fig. 6. HSE regulated IL-4, IL-5, and histamine in PM-exacerbated CARAS mice. (A,B) Levels of IL-4 and IL-5 in NLF. (C,D) Levels of IL-4 and IL-5 in BALF. (E) Levels of histamine in BALF. (G) Levels of IL-33 in BALF. All levels were determined using ELISA kits. Values are presented as mean ± SEM. Significant differences at ###p<0.001 and ##p<0.01 compared with the control group; ***p<0.001, **p<0.01, and *p<0.05 compared with the CARAS/PM group. CARAS, combined allergic rhinitis and asthma syndrome; Dexa, dexamethasone; HSE, Hydrangea serrata extract; OVA, ovalbumin; PM, particulate matter.

Article Snippet: Thus, IL-33 in BALF was measured using an ELISA kit (M33000, R&D Systems Inc., Minneapolis, MN, USA) according to the manufacturer’s guidelines.

Techniques: Enzyme-linked Immunosorbent Assay, Control