Journal: iScience

Article Title: Highly immunogenic DNA/LION nanocarrier vaccine potently activates lymph nodes inducing long-lasting immunity in macaques

doi: 10.1016/j.isci.2025.112232

Figure Lengend Snippet: DNA/LION vaccine induces durable T cell responses in rhesus macaques (A and B) Spike-specific T cell responses were measured in PBMCs over time by flow cytometry in (A) study 1, as IFN-γ + T cells, and (B) study 2, as cytokine + (IFN-γ, TNF-α single- and/or double-positive) T cells. Scatterplots with median line and minimum and maximum values are shown. The pie chart depicts the frequency of spike-specific IFN-γ, TNF-α single- and/or double-positive T cells. (C) Comparison of CD4 + and CD8 + IFN-γ + spike-specific T cell responses induced by DNA/LION (study 1, group 1) and by DNA(IM) prime followed by DNA/LION boost (study 1, group 2) and a historical study examining spike DNA vaccination by EP alone ( n = 4) or co-immunized with adjuvanted spike protein ( n = 4). (D) Frequency of spike-specific cytokine + T cells CD4 + and CD8 + T cells at the day of V4 and 2 and 3 weeks later (study 1). Bars indicate mean values. p values from generalized estimating equation (GEE) and are defined as ≤0.05, ∗; ≤0.01, ∗∗; ≤0.001, ∗∗∗; ≤0.0001,∗∗∗∗. (E) Frequency of cytokine + spike-specific CD8 + T CM (CD28 + CD95 + ) and T EM (CD28 − CD95 + CCR7 − ) after V4. Scatterplots with mean values are shown. (F) Proportion of cytokine + CD8 + T cells expressing EOMES and/or T-bet (mean of 8 animals). (G) Manually gated spike-specific cytokine + CD107a + CD8 + T CM and T EM cells (V4 [blue], V4w2 [red], V4w3 [green]) were overlaid in histograms depicting expression of EOMES, T-bet, GzmB, and Ki67 (mean fluorescent intensity [MFI] as mean of 8 animals at each time point). Gray dotted line marks the border between the negative and positive population.

Article Snippet: Mouse anti-human CD107a (LAMP-1) Monoclonal Antibody (eBioH4A3), PE, eBioscience , Thermo Fisher Scientific , Cat#12-1079-42; RRID: AB_10853326.

Techniques: Flow Cytometry, Comparison, Expressing

Journal: iScience

Article Title: Highly immunogenic DNA/LION nanocarrier vaccine potently activates lymph nodes inducing long-lasting immunity in macaques

doi: 10.1016/j.isci.2025.112232

Figure Lengend Snippet:

Article Snippet: Mouse anti-human CD107a (LAMP-1) Monoclonal Antibody (eBioH4A3), PE, eBioscience , Thermo Fisher Scientific , Cat#12-1079-42; RRID: AB_10853326.

Techniques: Staining, Clinical Proteomics, Recombinant, Binding Assay, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Software

Journal: bioRxiv

Article Title: Fc-dependent protective efficacy of non-inhibitory antibodies targeting influenza A virus neuraminidase is limited by epitope availability

doi: 10.1101/2024.09.03.611041

Figure Lengend Snippet: (a-d) The ability of the mAbs to induce activation of NK92 cells was determined following the incubation of the cells in plates coated with N1 CA09 or N2 VI75 in the absence or presence of the mAbs. The amount of activated cells was measured as CD107a + cells in flow cytometry and graphed as a percentage of the total NK cells (a and c). Binding of the mAbs to the NAs was measured in ELISA (b and d). (e to g) The induction of phagocytosis and complement deposition by the mAbs was determined in flow cytometry-based assays using microspheres. (e) Phagocytosis of microsphere-based immune complexes containing N1 WI13 and the mAbs by THP-1 monocytes was measured by flow cytometry. The phagoscores were calculated based on the percentage of THP-1 cells positive for the uptake of microspheres and the gMFI of that population as detailed in the Materials and Methods section. (f) Deposition of complement protein C3 on the microsphere-based immune complexes was measured by flow cytometry. (g) mAb binding to the N1-coated microspheres measured by flow cytometry. Data shown represent the mean ±SD of in total four replicates from two independent experiments combined.

Article Snippet: NK92 cells were diluted to 10 cells/ml in OptiMEM reduced serum medium (Gibco) supplemented with 1x Protein Transport Inhibitor Cocktail (Invitrogen) and V450 mouse anti-human CD107a (BD Biosciences; 1:400).

Techniques: Activation Assay, Incubation, Flow Cytometry, Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Fc-dependent protective efficacy of non-inhibitory antibodies targeting influenza A virus neuraminidase is limited by epitope availability

doi: 10.1101/2024.09.03.611041

Figure Lengend Snippet: 21H8 produced with reduced fucosylation (21H8ΔF) for enhanced FcγR engagement or mutated to abolish FcγR engagement (21H8 LALA-PG). Binding of the 21H8 variants against recombinant soluble N1 WI13 (a) and H1N1pdm09 virus (c) was quantified in ELISA. (b and d) The ability of the modified antibodies to activate NK cells upon binding to recombinant soluble N1 WI13 (b) or H1N1pmd09 virus (d) was quantified in flow cytometry as the percentage of CD107a + NK cells. Representative data of two independent experiments are shown (mean ±SD of two replicate measurements).

Article Snippet: NK92 cells were diluted to 10 cells/ml in OptiMEM reduced serum medium (Gibco) supplemented with 1x Protein Transport Inhibitor Cocktail (Invitrogen) and V450 mouse anti-human CD107a (BD Biosciences; 1:400).

Techniques: Produced, Binding Assay, Recombinant, Virus, Enzyme-linked Immunosorbent Assay, Modification, Flow Cytometry