Journal: PLoS ONE

Article Title: IL-12 and GM-CSF in DNA/MVA Immunizations against HIV-1 CRF12_BF Nef Induced T-Cell Responses With an Enhanced Magnitude, Breadth and Quality

doi: 10.1371/journal.pone.0037801

Figure Lengend Snippet: A ) Functional avidity of the BF33- and B84-specific T-cells: Groups of 4 mice received the immunization schemes depicted in for the group 1 (NefBF alone) and group 2 (plus IL-12 during the priming phase). Nine days after the MVA boost, T- cell avidity against BF33 and B84 peptides was evaluated using serial dilutions of the peptides. Peptide concentrations used ranged from 20 to 0.002 µg/ml. SD 50 : sensitizing dose of peptides required to yield 50% maximal T-cell triggering of IFN-γ production. B ) Spleen cells from the indicated group were evaluated for their proliferative capacity against the BF33 peptide, and the identification of the specific T-cell subset was performed. For this, cells were dyed with CFSE and stimulated with BF33 peptide during 5 days and then staining with surface antibodies was performed (CD3, CD4 and CD8). i- Representative dot plots. Numbers inside the graphs represent the average values for duplicated samples. ii- Bars represent the average of duplicated samples plus SD. Background values obtained in cell-cultures stimulated with RPMIc, were subtracted in each case. T test was applied, *p<0.05, * * p<0.005.

Article Snippet: In these experiments, the responses found in the control vs the IL-12 group were compared, analyzing the avidity of the T-cells against the BF33 (same sequence as the immunogen vectors) or the B84 peptide (with 3 aa changes, see ), in order to define if the recognition pattern differed between homologous or heterologous responses.

Techniques: Functional Assay, Staining

Journal: PLoS ONE

Article Title: IL-12 and GM-CSF in DNA/MVA Immunizations against HIV-1 CRF12_BF Nef Induced T-Cell Responses With an Enhanced Magnitude, Breadth and Quality

doi: 10.1371/journal.pone.0037801

Figure Lengend Snippet: Results summary.

Article Snippet: In these experiments, the responses found in the control vs the IL-12 group were compared, analyzing the avidity of the T-cells against the BF33 (same sequence as the immunogen vectors) or the B84 peptide (with 3 aa changes, see ), in order to define if the recognition pattern differed between homologous or heterologous responses.

Techniques: Control

Journal: Nature Communications

Article Title: Tissue-intrinsic beta-catenin signals antagonize Nodal-driven anterior visceral endoderm differentiation

doi: 10.1038/s41467-024-49380-0

Figure Lengend Snippet: a Experimental approach to determine clonal composition of AVE nests. b Expression of clonal labels (red, cyan) and Cer1:H2B-Venus reporter (yellow) in cultures differentiated as in a . Insets on the right show examples of Cer1:H2B-Venus-expressing nests with a single clonal label (top, 13/30 nests), or with multiple labels (bottom, 17/30 nests). One out of n = 2 independent experiments shown. c Immunostaining for OTX2 (magenta) and H2B-Venus (yellow) of Cer1:H2B-Venus reporter cells differentiated for 3 days after an extended doxycycline pulse with 50 ng/ml ActivinA (AA), together with 3 µM Chir99021 (Chi), 20 µM XAV939 (XAV), or 2 µM IWP2 as indicated. One out of n = 3 independent experiments shown. d Flow cytometry of cells differentiated and stained as in c . e Mean percentage of Cer1:H2B-Venus; OTX2 double-positive cells differentiated as in c . n = 4 independent experiments, error bars indicate SD. ** p < 0.001 for AA vs. AA + Chi and AA vs. AA + XAV, and p = 0.0045 for AA vs. AA + IWP2 (two-tailed, unpaired t -test). f Same as e but showing percentage of OTX2-positive cells. ** p < 0.001 for AA vs. AA + Chi and p = 0.0001 for AA vs. AA + XAV, * p = 0.0434 for AA vs. AA + IWP2 (two-tailed, unpaired t -test). Scale bars: 200 µm (( b ) overview); 20 µm (( b ) inset); 500 µm (( c ) overview); 50 µm (( c ) inset). Source data for e , f are provided in the Source Data file.

Article Snippet: To test the functionality of the putative AVE, we adopted a protocol to trigger mesoderm differentiation in ESC aggregates , consisting of a 24 hour treatment with 3 μM Chir99021 (Chi) on day 2, followed by analysis 24 hours after the end of the Chi pulse (Supplementary Fig. ).

Techniques: Expressing, Immunostaining, Flow Cytometry, Staining, Two Tailed Test