tlr 7 8 agonist  (InvivoGen)

Journal: PLOS Pathogens

Article Title: Peculiar transcriptional reprogramming with functional impairment of dendritic cells upon exposure to transformed HTLV-1-infected cells

doi: 10.1371/journal.ppat.1012555

Figure Lengend Snippet: a. Schematic representation of the experimental design. MDDCs were exposed to uninfected (Jurkat, green) or HTLV-1-infected T cells (C91-PL, blue) for 24h, before restimulation with LPS or R848 for an additional 24h. b. Flow cytometry analysis after CD11c and CD86 staining, in LPS- ( left ) or R848-restimulated MDDCs ( right ) pre-exposed to Jurkat (red) or C91-PL (dark blue) cells. Data are represented as the normalized MFI of CD86, with the MFI in restimulated Jurkat-pre-exposed MDDCs set to 100 (n = 30 or 10 independent experiments, respectively). Presented data are a subset of , respectively and were analysed with Kruskal-Wallis test or ordinary one-way ANOVA, respectively as described in and Tables c. Supernatant from the indicated cocultures was collected after LPS restimulation, and TNF-α ( left ) and IFN-I ( right ) concentrations were quantified for n = 3 or 10 independent experiments, respectively. Presented data are a subset of and were analysed with RM one-way ANOVA or ordinary one-way ANOVA, respectively as described in and Tables. d. Schematic drawing summarizing the results from Figs and . The drawing was created using BioRender.com .

Article Snippet: Toll-like receptor (TLR)-4 agonist (LPS, tlrl-3pelps; 1μg/mL) and TLR-7/8 agonist (R848, tlrl-r848; 3μg/mL) were purchased from Invivogen.

Techniques: Infection, Flow Cytometry, Staining

Journal: iScience

Article Title: Functional diversification of innate and inflammatory immune responses mediated by antibody fragment crystallizable activities against SARS-CoV-2

doi: 10.1016/j.isci.2024.109703

Figure Lengend Snippet: Rate of cell mortality in SARS-CoV-2-treated PBMC is not affected by adding of J08 and F05 nAbs

Article Snippet: Cells were also treated, where indicated, with the TLR-7/8 agonist Resiquimod (R848, 5 μM, Invivogen) and the TLR-9 ligand type C CpG (3 μg/ml, Invivogen) in presence or absence of increasing doses of J08 or F05 nAbs.

Techniques: Cell Culture

Journal: iScience

Article Title: Functional diversification of innate and inflammatory immune responses mediated by antibody fragment crystallizable activities against SARS-CoV-2

doi: 10.1016/j.isci.2024.109703

Figure Lengend Snippet: FcγR crosslinking with J08 nAb immune complexes and Fc-mediated viral entry in PBMC (A) Schematic representation of the in vitro experimental setting used to mimc FcγR crosslinking. Biotynilated recombinant trimeric Spike (B-Spike) protein was coupled to streptavidin (SA)-labeled magnetic beads. Immune complexes (IC) were formed between anti-SARS-CoV-2 Spike neutralizing IgG1 antibody J08 in the wild-type (WT) version at the neutralizing dose and trimeric Spike coupled to SA-magnetic beads. IC were added to peripheral blood mononuclear cells (PBMC) to crosslink Fcγ Receptors (FcγR) on different FcγR-expressing PBMC subsets. (B‒E) PBMC isolated from healthy volunteers ( n = 3) were left untreated (not stimulated, ns) or stimulated with J08 WT neutralizing antibody (nAb) and biotinylated recombinant (rec) trimeric Spike protein alone or coupled to SA-magnetic beads. As positive control biotynilated goat anti-human CD16, CD32 and CD64 antibodies (anti-FcγR Abs) were also coupled to SA-magnetic beads and then mixed together in equal quantity to crosslink all the FcγR expressed by PBMC subsets. Biotynilated goat anti-human immunoglobulin G (Isotype Ab) coupled to SA-magnetic beads was used as a negative control. Cells were also stimulated with the toll-like receptor (TLR)-7/8 agonist R848 and the TLR-9 agonist class C CpG. Production of interferon-alphas (IFN-αs) (B), interleukin (IL)-6 (C), tumor necrosis factor (TNF)-α (D) and IL-8 (E) was measured in culture supernatants. (F and G) PBMC were adsorbed for 1 h with virus-like particles (VLP) expressing green-fluorescent protein (GFP) pseudotyped with membrane-tethered SARS-CoV-2 (D614G) Spike (S) protein alone or immunocomplexed with J08 nAbs in the WT or MUT versions. (F) Representative dot plots of the different experimental conditions derived from 1 experiment, out of 3 independently performed, are shown. (G) Results shown are the mean values of the percentage (%) of GFP + cells in total single live PBMC derived from the 3 experiments independently performed. p -values were calculated by two-way ANOVA and assigned as follows: ∗ ≤0.05; ∗∗∗∗ ≤0.0001.

Article Snippet: Cells were also treated, where indicated, with the TLR-7/8 agonist Resiquimod (R848, 5 μM, Invivogen) and the TLR-9 ligand type C CpG (3 μg/ml, Invivogen) in presence or absence of increasing doses of J08 or F05 nAbs.

Techniques: In Vitro, Recombinant, Labeling, Magnetic Beads, Expressing, Isolation, Positive Control, Negative Control, Virus, Membrane, Derivative Assay

Journal: iScience

Article Title: Functional diversification of innate and inflammatory immune responses mediated by antibody fragment crystallizable activities against SARS-CoV-2

doi: 10.1016/j.isci.2024.109703

Figure Lengend Snippet:

Article Snippet: Cells were also treated, where indicated, with the TLR-7/8 agonist Resiquimod (R848, 5 μM, Invivogen) and the TLR-9 ligand type C CpG (3 μg/ml, Invivogen) in presence or absence of increasing doses of J08 or F05 nAbs.

Techniques: Affinity Purification, Virus, Variant Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Transfection, Software, FCAP Assay, Cytometry