Journal: Biomolecules

Article Title: Machine Learning-Guided Prediction of Antigen-Reactive In Silico Clonotypes Based on Changes in Clonal Abundance through Bio-Panning

doi: 10.3390/biom10030421

Figure Lengend Snippet: Reactivity of phage-displayed scFv clones in phage ELISA. Binding reactivity of 15 unique clones identified from the AR library and 16 unique clones from the NR library are shown. Wells in microtiter plates were either coated with recombinant mouse c-Met or just blocked with 3% BSA in PBS. Phage clones, HRP-conjugated anti-M13 antibody, and HRP substrate solution were added sequentially with intermittent washing.

Article Snippet: After a round of bio-panning using recombinant mouse c-Met (50622-M08H; Sino Biological)-conjugated magnetic beads (Dynabeads 14302D; Invitrogen) and washing once with 0.05% tween in PBS, 96 phage clones were randomly rescued from each AR and NR library and subjected to phage ELISA, as described above.

Techniques: Clone Assay, Enzyme-linked Immunosorbent Assay, Binding Assay, Recombinant

Journal: Journal of Innate Immunity

Article Title: Impairment in Natural Killer Cells Editing of Immature Dendritic Cells by Infection with a Virulent Trypanosoma cruzi Population

doi: 10.1159/000350242

Figure Lengend Snippet: Frequency, activation and cytokine production by NK cells following T. cruzi infection. C3H/HeN mice were infected with the HV and LV T. cruzi populations and sham infected. PBMC (blood) and spleen cell suspensions (spleen) obtained at different time points after infection. Frequency of DX5+/CD3- cells and expression of CD69 by DX5+/CD3- cells were evaluated in PMBC (a) and spleen cell suspensions (b). Ex vivo intracellular production of IFN-γ and IL-10 by DX5+/CD3- cells from PBMC fractions (c) and spleen cell suspensions (d) 18 h.p.i. Dot plots show results from 1 representative experiment of 3 each using a pool of 3-5 mice per group. Bar graphs show results obtained from 3 independent experiments with 3-5 pooled mice per group (means ± SEM). ANOVA (Bonferroni's post hoc test). * p < 0.05 sham vs. LV and HV, # p < 0.05 HV vs. sham and LV.

Article Snippet: Purification of NK Cells DX5+ cells were isolated from nonadherent spleen cell fractions by magnetic sorting using biotinylated anti-mouse CD49/pan-NK cell (DX5) monoclonal antibodies bound to streptavidin-coated magnetic beads and miniMACS columns (Miltenyi Biotech GmbH, Germany) according to the manufacturer's protocol.

Techniques: Activation Assay, Infection, Expressing, Ex Vivo

Journal: Journal of Innate Immunity

Article Title: Impairment in Natural Killer Cells Editing of Immature Dendritic Cells by Infection with a Virulent Trypanosoma cruzi Population

doi: 10.1159/000350242

Figure Lengend Snippet: NK cell-mediated lysis of YAC-1 cells and iDCs upon T. cruzi infection. a Lysis of YAC-1 cells by splenic NK cells from T. cruzi HV, LV or sham-infected (18 h.p.i.) and LPS (100 µg/mouse) inoculated C3H/HeN mice was evaluated. YAC-1 cells (5 × 104) were cultured with freshly purified splenic NK cells at a NK/YAC-1 ratio of 5/1. Left panel shows CFSE-labeled YAC-1 cells and middle panel shows PI uptake by CFSE+ cells gated in R. Results from a representative experiment with 3-5 pooled mice per group. Right panel show results of 3 independent experiments with 3-5 pooled mice per group (means ± SEM). ANOVA (Bonferroni's post hoc test). * p < 0.05 sham vs. LV and HV. b iDCs (5 × 104) were cultured with freshly purified spleen NK cells from HV, LV and sham-infected (18 h.p.i.) C3H/HeN mice. Left panel shows CFSE-labeled iDCs and middle panel shows PI uptake by CFSE+ cells gated in R. Results from a representative experiment with 3-5 pooled mice per group. Right panel shows results of 3 independent experiments with 3-5 pooled mice per group (means ± SEM). ANOVA (Bonferroni's post hoc test) * p < 0.05 sham vs. LV and HV.

Article Snippet: Purification of NK Cells DX5+ cells were isolated from nonadherent spleen cell fractions by magnetic sorting using biotinylated anti-mouse CD49/pan-NK cell (DX5) monoclonal antibodies bound to streptavidin-coated magnetic beads and miniMACS columns (Miltenyi Biotech GmbH, Germany) according to the manufacturer's protocol.

Techniques: Lysis, Infection, Cell Culture, Purification, Labeling

Journal: Journal of Innate Immunity

Article Title: Impairment in Natural Killer Cells Editing of Immature Dendritic Cells by Infection with a Virulent Trypanosoma cruzi Population

doi: 10.1159/000350242

Figure Lengend Snippet: IL-10 mediates NK cell cytotoxicity on iDCs. Mice were infected with HV T. cruzi. a iDCs (5 × 104 per well) were cultured with freshly purified spleen NK cells (NK/DC ratio: 2/1) from HV (■) and sham-infected (□) C3H/HeN male mice (18 h.p.i.) in the presence of an IL-10R blocking antibody (Ab; 40 µg/ml, clone 1B1.3A) or rat IgG isotype control (HPRN). Results are expressed as percent lysis from 2 independent experiments with 4-5 pooled mice per group (means ± SEM). ANOVA (Bonferroni's post hoc test). * p< 0.05 vs. sham isotype. b iDCs (5 × 104/well) were cultured with freshly purified spleen NK cells (NK/DC ratio: 2/1) from IL-10KO mice of Balb/c background or wild-type (WT) mice (18 h.p.i.). Results are expressed as percent lysis from 2 independent experiments with 4-5 pooled mice per group (means ± SEM). ANOVA (Bonferroni's post hoc test). * p< 0.05 vs. HV WT.

Article Snippet: Purification of NK Cells DX5+ cells were isolated from nonadherent spleen cell fractions by magnetic sorting using biotinylated anti-mouse CD49/pan-NK cell (DX5) monoclonal antibodies bound to streptavidin-coated magnetic beads and miniMACS columns (Miltenyi Biotech GmbH, Germany) according to the manufacturer's protocol.

Techniques: Infection, Cell Culture, Purification, Blocking Assay, Control, Lysis