Journal: Antibodies

Article Title: Functional Activity of Cytokine-Induced Killer Cells Enhanced by CAR-CD19 Modification or by Soluble Bispecific Antibody Blinatumomab

doi: 10.3390/antib13030071

Figure Lengend Snippet: In vitro activity. ( A ) Comparison of the killing activity in vitro using the calcein release assay. CIK (blue bars), CIK in presence of 10 ng/mL blinatumomab (red bars), CARCIK-MNZ (pink bars), and CARCIK-BG2 (green bars) at the end of culture were used as cytotoxic effector cells against the REH cell line at a 10:1, 3:1, and 1:1 E:T ratio. Percentage target lysis is shown as mean and standard deviation of six experiments. ( B ) Comparison of the proliferation ability in vitro (proliferation index) measured by flow cytometry on CFSE-stained effector cells, after co-culture with the target cell line REH, at a 10:1, 1:1, and 1:10 E:T ratio, for 4 days. Proliferation indexes are shown as mean and standard deviation of four experiments. ( C , D ) IFN-γ and IL-2 production was determined by intracytoplasmic flow cytometry after 6 h co-culture at 1:1 E:T ratio with REH cell line. Percentages of positive cells are shown as mean and standard deviation of five experiments. ( E , F ) The NFAT and NF-kB upon stimulation was measured by co-transfection of Lucia luciferase reporter plasmids into HuT 78 cell line wild type alone (as CIK, blue bars), or stimulated with 10 ng/mL blinatumomab (red bars), or stably transfected with CAR-MNZ (pink bars) or with CAR-BG2 (green bars) and co-cultured with REH cell line for 24 h. Relative luminescence units (RLUs) are shown as mean and standard deviation of five experiments. Columns represent the mean and bars the standard deviation (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: To assess NF-kB and NFAT signaling, 2.5 μg NFAT or NF-kB inducible secreted luciferase reporter plasmids (pNifty3-T-Lucia and pNifty3-N-Lucia, respectively) (InvivoGen, San Diego, CA, USA) were transfected into CAR + or wild type HuT 78, using the nucleofector kit V and Amaxa IIb nucleofector device (Lonza).

Techniques: In Vitro, Activity Assay, Comparison, Release Assay, Lysis, Standard Deviation, Flow Cytometry, Staining, Co-Culture Assay, Cotransfection, Luciferase, Stable Transfection, Transfection, Cell Culture

Journal: Microbiome Research Reports

Article Title: Galacto-oligosaccharides alone and combined with lactoferrin impact the Kenyan infant gut microbiota and epithelial barrier integrity during iron supplementation in vitro

doi: 10.20517/mrr.2024.34

Figure Lengend Snippet: Overview of mammalian cell model setup using a transwell system to assess the impact of differently treated PolyFermS supernatant on the epithelial barrier, pathogen infection, and inflammatory response. TEER: Transepithelial electrical resistance; MOI: multiplicity of infection; LDH: lactate dehydrogenase; ETEC: enterotoxigenic Escherichia coli ; NFκB: nuclear factor kappa B; SEAP: secreted embryonic alkaline phosphatase.

Article Snippet: THP-1 Blue cells were transfected with a nuclear factor kappa B (NFκB)-inducible secreted embryonic alkaline phosphatase (SEAP) reporter construct (Invivogen), which enables the screening of pro-inflammatory NFκB activation.

Techniques: Infection