Journal: PLOS One

Article Title: Rabies vaccination induces a CD4 + T EM and CD4 + CD8 + T EMRA T H 1 phenotype in dogs

doi: 10.1371/journal.pone.0323823

Figure Lengend Snippet: (A) Total IgG titers against RABV-G and RABV-N on the day of a rabies vaccine booster (Day 0) or 7-14 days after vaccine administration. The dashed line was determined by the antibody titer from an unvaccinated puppy (Dog 11, ) and was used as a normalization control for each experiment ( n = 10, paired two-tailed Wilcoxon test; ns, not significant, ** p < 0.01; two independent experiments). (B) Representative IFNγ ELISpot images of mock-, RABV-N-, and RABV-G- stimulated PBMCs performed in triplicate. (C) Quantification of the data represented in (B) ( n = 6; one-way ANOVA and post hoc Tukey’s test; ns, not significant, * p < 0.05, ** p < 0.01; three independent experiments).

Article Snippet: A canine IFNγ ELISpot Development Module (SEL781, R&D Systems) was used per the manufacturer’s protocol with slight modifications.

Techniques: Control, Two Tailed Test, Enzyme-linked Immunospot

Journal: PLOS One

Article Title: Rabies vaccination induces a CD4 + T EM and CD4 + CD8 + T EMRA T H 1 phenotype in dogs

doi: 10.1371/journal.pone.0323823

Figure Lengend Snippet: (A) Representative histogram plots compare the mock control (black outline) to RABV-G-stimulated T cell subsets: CD4 + (top row, blue), DP (middle row, orange), and CD8 + (bottom row, black), with % proliferated (CTFR LO ) shown within the plot. (B) Quantification of (A) (mean ± SEM; n = 6/experiment; two-way ANOVA with multiple comparisons **** p < 0.0001; 6 independent experiments). Data was normalized to controls. (C) Representative histograms of IFNγ (left) and IL-17A (right) expression by CD8 + , CD4 + , or DP T cells stimulated with RABV-G after 5 days. (D) Mean fluorescence intensity (MFI) of IFNγ (left) and IL-17A (right) for CD4 + T cells, DP T cells, and CD8 + T cells (mean ± SEM; n = 6/experiment; two-way ANOVA with multiple comparisons; ns, not significant, * p < 0.05, *** p < 0.001, **** p < 0.0001; 6 independent experiments). MFI was normalized to controls.

Article Snippet: A canine IFNγ ELISpot Development Module (SEL781, R&D Systems) was used per the manufacturer’s protocol with slight modifications.

Techniques: Control, Expressing, Fluorescence

Journal: PLOS One

Article Title: Rabies vaccination induces a CD4 + T EM and CD4 + CD8 + T EMRA T H 1 phenotype in dogs

doi: 10.1371/journal.pone.0323823

Figure Lengend Snippet: (A) Gating strategy and representative contour plots for lineage-defining subsets between CTFR LO and CTFR HI T cells to differentiate T H 1 (T-bet + IFNγ + ), T H 17 (IL-17A + ), T H 1/T H 17 (T-bet + IL-17A + ), pathogenic T H 1/T H 17 (T-bet + IL-17A + IFNγ + ), and Tregs (FOXP3 + ). (B) Quantification of (A) (mean ± SEM, n = 22 samples, paired two-tailed Wilcoxon test; * p < 0.05, *** p < 0.001, **** p < 0.0001; 4 independent experiments).

Article Snippet: A canine IFNγ ELISpot Development Module (SEL781, R&D Systems) was used per the manufacturer’s protocol with slight modifications.

Techniques: Two Tailed Test

Journal: Scientific Reports

Article Title: Immunological responses following administration of a genotype 1a/1b/2/3a quadrivalent HCV VLP vaccine

doi: 10.1038/s41598-018-24762-9

Figure Lengend Snippet: B cell responses and total IgG secretion following immunization with quadrivalent vaccines. BALB/c mice (n = 5/group) were immunized subcutaneously at the base of the tail with PBS alone (control) or 80 μg of quadrivalent VLP. Splenocytes were harvested on day 21 and frequencies of total HCV specific antibody secreting cells (ASCs) in each group (PBS, or vaccine combined with Alum, CFA or Montanide) ( A ) and total IgG ( B ) were determined by B cell ELISpot assay. The y-axis shows the antibody secreting cell (ASC) number/million splenocytes. Individual animals are presented for each group, with the mean value being represented by the horizontal bar.

Article Snippet: For the detection of Granzyme B-secreting cells, a Mouse Granzyme B ELISpot Development Module together with reagents from an ELISpot Blue Color Module (R&D Systems, Minneapolis, USA) including a Granzyme B positive control were used according to the manufacturer’s instructions.

Techniques: Vaccines, Control, Enzyme-linked Immunospot

Journal: Scientific Reports

Article Title: Immunological responses following administration of a genotype 1a/1b/2/3a quadrivalent HCV VLP vaccine

doi: 10.1038/s41598-018-24762-9

Figure Lengend Snippet: IFNγ T cell responses following immunization with quadrivalent vaccines. ( A ) BALB/c mice (n = 5/group) were immunized subcutaneously at the base of the tail with the quadrivalent VLP combinations as shown. Frequencies of IFNγ secreting cells before and after restimulation were determined by ELISpot assay. ( B ) HHD mice were vaccinated with OT2 Pam2Cys GIL, HCV VLPs or PBS and frequencies of IFNγ secreting cells were determined by ELISpot after stimulation with GILGFVFTL, HCV VLP, HCVcore 132_−140 DLMGYIPLV (DLM), HCVcore 35–44 YLLPRRGPRL (YLL) or the irrelevant HCV NS5B 2594–2602 peptide. ( C ) T cells from BALB/c mice vaccinated with quadrivalent VLP alone were further purified from spleen using Miltenyi Biotec columns and CD4 (L3T4) and CD8a (Ly-2) microbeads and analysed by ELISpot assay. The mean value and standard deviation is shown for each treatment group.

Article Snippet: For the detection of Granzyme B-secreting cells, a Mouse Granzyme B ELISpot Development Module together with reagents from an ELISpot Blue Color Module (R&D Systems, Minneapolis, USA) including a Granzyme B positive control were used according to the manufacturer’s instructions.

Techniques: Vaccines, Enzyme-linked Immunospot, Purification, Standard Deviation

Journal: Scientific Reports

Article Title: Immunological responses following administration of a genotype 1a/1b/2/3a quadrivalent HCV VLP vaccine

doi: 10.1038/s41598-018-24762-9

Figure Lengend Snippet: Granzyme B responses following immunization with quadrivalent vaccines. BALB/c mice (n = 5/group) were immunized subcutaneously at the base of the tail with the quadrivalent VLP combinations as shown. Splenocytes were harvested on day 21 and frequencies of Granzyme ( B) responses before and after restimulation were determined by ELISpot assay. The mean value and standard deviation is shown for each treatment group.

Article Snippet: For the detection of Granzyme B-secreting cells, a Mouse Granzyme B ELISpot Development Module together with reagents from an ELISpot Blue Color Module (R&D Systems, Minneapolis, USA) including a Granzyme B positive control were used according to the manufacturer’s instructions.

Techniques: Vaccines, Enzyme-linked Immunospot, Standard Deviation