This clarification of the mechanisms underlying the CD8 + CD28 − expansion in HIV-induced pathogenesis may have important therapeutic implications.Dalod, MarcLaboratoire d'Immunologie des pathologies infectieuses et tumorales, Unité 445 INSERM, Institut Cochin de Génétique Moléculaire...
FMO, fluorescence minus one control. Additional file 8: Figure S3. The T-cell differentiation stage impacts the circulating CD28/PD-1 subset distribution and changes as a function of the activation interval. A Unstimulated ex vivo PBMC from NSCLC (n = 22), melanoma (n = 10), ...
Frequency, differentiation stage and functionality of peripheral CD8+T-cell subsets defined by PD1 and/or CD28 expression.ARepresentative flow cytometry plots showing PD1vsCD28 staining, in ex vivo CD8+T cells from PBMC of patients with three different tumors, as indicated.BPercentage of circulating ...
T cells from healthy donor peripheral blood mononuclear cells were purified by negative selection using a T cell isolation kit (Miltenyi) and subsequently activated with CD3/CD28 Dynabeads (Thermo Fisher Scientific) at a cell-to-bead ratio of 1:3. A lentiviral vector carrying a previously ...
The majority of immune-oncology assays use pre-stimulated effector T cells with anti-CD3 and anti-CD28 antibodies, relatively high doses of cytokines (e.g. IL-2, IFNγ) or unspecific stimulation with concanavalin A or PMA and ionomycin, which could bias the results when testing immunotherapeuti...
15 Briefly, 4 × 106 PBMCs were stimulated once with 10 μg/mL peptide and 0.5 μg/mL anti-CD28 (BD Biosciences) and fed every 3 days with complete medium containing 20 U/mL recombinant interleukin (IL)-2 (Miltenyi Biotec, Bergisch Gladbach, Germany). Peptide-specific CD8+ T-cell ...
(Supplementary Fig.1a, b). In an in vitro system mimicking different states of human CD8 T cells28, a highYthdf2mRNA level was observed during 3–48 h of anti-CD3/CD28 stimulation, but its expression decreased at later time points of activation as well as towards exhaustion-like and ...
(a) Schematic of acute (left) and chronic stimulation (right) with CD3/CD28 dynabeads. (b) Average percentage of positive cells (top panel: PD1 (p = 0.02), LAG3 (p = 0.06), TIGIT (p = 0.03), and TIM3 (ns)) and MFI (bottom panel: PD1 (p = 0.02), LAG3 ...
CD8+ T cells were activated by plating cells into a plate pre-coated with 1 μg/mL anti-CD3 (clone OKT3; BioLegend) and anti-CD28 (clone CD28.2; BioLegend) and expanded in complete RPMI medium supplemented with 100 U/mL recombinant human IL-2 (PeproTech). T cells were used ...
Gating strategy of survivin positive cells show its separation from the fluorescence minus one (FMO) staining control. G: Percentage of survivin+ cells expressing CD93 and IgM. Boxes represent interquartile range, horizontal lines indicate median, and whiskers indicate min to max range. Comparison ...