Journal: PLoS ONE

Article Title: MyD88/CD40 Genetic Adjuvant Function in Cutaneous Atypical Antigen-Presenting Cells Contributes to DNA Vaccine Immunogenicity

doi: 10.1371/journal.pone.0164547

Figure Lengend Snippet: (A) NIH3T3 or (B) MPEK stable cells lines expressing either GFP alone (neg. control), OVA or MC.OVA were incubated for 60 hours. MC.OVA +rim had 10 nM rim present in the culture. After 60 hours, cells were harvested and MHC-I expression was analyzed by flow cytometry. (C-E) MPEK stable cells surface expression of CD80, CD86, and CD40. MFI values of live cells are reported relative to the mean MFI of negative control cells, transduced with an empty-vector, in each respective sample. (F) CD3 + CD8 + OT-1 or WT T cells were purified by MACs and labeled with CellTrace violet (CTV) dye. Then 5 x 10 4 CD3 + CD8 + CTV dye-labeled OT-1 or WT T cells were incubated with 5 x 10 4 B6-MPEK cells for 60 hours. By flow cytometry, live CTV + cells were analyzed for CTV dye dilution as a measure of cell proliferation. (G) Representative histograms of live CTV + cells. Percentages represent the percent of cells proliferating. n = 3, *p<0.05, One-way ANOVA, Tukey correction for multiple comparisons.

Article Snippet: Cells were then labeled with CellTrace Violet (CTV) cell proliferation dye (ThermoFisher Scientific, Waltham, MA) at either a high (Hi) concentration (5 μM) or low (Lo) concentration (0.5 μM) for 25 minutes at 37°C, per the manufacturer’s recommendations.

Techniques: Expressing, Control, Incubation, Flow Cytometry, Negative Control, Transduction, Plasmid Preparation, Purification, Labeling

Journal: Microorganisms

Article Title: A Weakened Immune Response to Synthetic Exo-Peptides Predicts a Potential Biosecurity Risk in the Retrieval of Exo-Microorganisms

doi: 10.3390/microorganisms8071066

Figure Lengend Snippet: Iva- or Aib promote proliferation of OT-I cells. Cell proliferation was assessed by measuring dilution of CellTrace Violet dye over 72 h following stimulation with the test peptides. ( A – L ) Representative images show a histogram shift ( A – F ) of unstimulated control cells (purple histogram) against cells stimulated with peptides (green). ( G – L ): Representative images show dot plots of activated (CD25 expressing) cells proliferating (decrease in CellTrace) cells when unstimulated (purple) or stimulated with peptides (green). The native ova-control peptides ( A , G ): short Ova 257-264 (SIINFEKL) or ( B , H ): longer Ova 248-269 ). Ova peptides substituted with exo-amino acids ( C , I ): Ova 248-269 (Iva) and ( E , K ): Ova 248-269 (Aib) or peptides substituted natural amino acids ( D , J ): Ova 248-269 (Val) and ( F , L ): Ova 248-269 (Ala). ( M ): Geometric Mean (CellTrace) of proliferating CD8 T cells. Proliferation with peptide Ova 248-269 (Iva) or Ova 248-269 (Aib) are statistical significant ( p -value < 0.01) compared to the control peptides Ova 248-269 ( p -value 0.0032 and p -value 0.0015) and control Ova 248-269 (Val) and Ova 248-269 (Ala), p -value 0.004 and p -value 0.0021, respectively. ( N ): Mean fluorescent Intensity (MFI) of proliferating CD8 T cells expressing CD25. Peptide Ova 248-269 (Iva) or Ova 248-269 (Aib) are statistical significant compared to the control peptides Ova 248-269 (**: p -value < 0.01; *: p -value < 0.1). Three technical replicas were performed for each of the three biological replicas.

Article Snippet: Activated lymphocytes were analysed for cell proliferation by measuring dilution of CellTrace Violet dye (450nm EX /450nm EM , Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Control, Expressing