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: Human Gene Therapy

Article Title: Evaluation of Adeno-Associated Viral Vectors for Liver-Directed Gene Transfer in Dogs

doi: 10.1089/hum.2010.194

Figure Lengend Snippet: Time course of T cell responses to GFP and capsid in peripheral blood mononuclear cells (PBMCs) and lymphocytes isolated from spleen and liver, determined by IFN-γ ELISpot assay. T cells were stimulated with GFP or peptide pools (A, B, C) corresponding to AAV2 and AAV8 capsid, respectively, and the number of IFN-γ-secreting cells in response to antigen stimulation was determined. T cell responses to GFP can be observed in the two animals that received AAV8 on day 27. SFC, spot-forming cells.

Article Snippet: Enzyme-linked immunospot (ELISpot) assays to detect canine interferon (IFN)-γ were performed with the canine IFN-γ ELISpot development module and ELISpot blue color module (R&D Systems, Minneapolis, MN), in accordance with the manufacturer's instructions.

Techniques: Isolation, Enzyme-linked Immunospot

Journal: Human Gene Therapy

Article Title: Evaluation of Adeno-Associated Viral Vectors for Liver-Directed Gene Transfer in Dogs

doi: 10.1089/hum.2010.194

Figure Lengend Snippet: Development of T cells against transgene and capsid in animal S468 determined by IFN-γ ELISpot. Top: Time course of T cell frequencies in PBMCs against GFP and AAV8 capsid (peptide pools 8A, 8B, 8C; control, C). Bottom: Frequencies of GFP and capsid-specific T cells isolated from spleen and liver compared with PBMCs on day 35 after vector administration.

Article Snippet: Enzyme-linked immunospot (ELISpot) assays to detect canine interferon (IFN)-γ were performed with the canine IFN-γ ELISpot development module and ELISpot blue color module (R&D Systems, Minneapolis, MN), in accordance with the manufacturer's instructions.

Techniques: Enzyme-linked Immunospot, Control, Isolation, Plasmid Preparation