Journal: PLoS Pathogens

Article Title: A Higher Activation Threshold of Memory CD8 + T Cells Has a Fitness Cost That Is Modified by TCR Affinity during Tuberculosis

doi: 10.1371/journal.ppat.1005380

Figure Lengend Snippet: (a) The TB10 tetramer + response enumerated by duel-tetramer staining in blood 1 week post-boost with TB10 4-11 or Ova 257-264 (control) vaccination. Numbers indicate the % of CD8 + T cells. (b) TB10 tetramer + responses from blood after TB10 4-11 or control (B8R 20-27 or Ova 257-264 ) vaccination at time points post-boost. (c) Representative plots showing CD62L, KLRG1, IL-7R, and CXCR3 expression by TB10 tetramer + CD8 + T cells from blood 1w after prime, and 1w or 8w after boost. (d) Ex vivo TB10 4-11 -stimulated production of IFNγ, TNF, or granzyme B from CD8 + T cells isolated from combined lungs, spleens, or LNs of TB10-vaccinated mice. (e) In vivo specific killing of targets coated with TB10 4-11 peptide. (f) In vivo specific killing of 1μM TB10 4-11 -coated targets vs. TB10-specific response. (g) TB10 tetramer responses of mice vaccinated with TB10 4-11 or the control peptide B8R 20-27 immediately prior, or 2w, 4w, or 12w after Mtb infection. **** p < 0.0001, *** p < 0.001, by two-way ANOVA with Sidak’s post test. Data are representative of 3–6 independent experiments, each with 4–6 mice per group.

Article Snippet: TB10.4 4−11 (IMYNYPAM), B8R 20-27 (TSYKFESV), “amphi-TB10” (MFVMFVQIMYNYPAM), and ovalbumin 257-264 (SIINFEKL) peptides were purchased from New England Peptides (Gardner, MA, USA) and reconstituted in DMSO (10mM).

Techniques: Staining, Expressing, Ex Vivo, Isolation, In Vivo, Infection

Journal: Virology

Article Title: HIV-1 Gag p17 presented as virus-like particles on the E2 scaffold from Geobacillus stearothermophilus induces sustained humoral and cellular immune responses in the absence of IFN? production by CD4+ T cells

doi: 10.1016/j.virol.2010.08.026

Figure Lengend Snippet: C57BL/6 recipient mice (5 per group) of CFSE-labelled OT-II T cells were immunized with E2wt particles, OVA323-339 E2 particles or soluble ovalbumin (OVA) plus poly (I:C) or PBS. 3 days after immunization, mice were sacrificed and the spleen cells analysed by flow cytometry for CFSE content (A–B), or restimulated in vitro with OVA323-339 synthetic peptide for IFNγ production analysis (C–D).

Article Snippet: The OVA 257-264 and OVA 323-339 peptides from chicken ovalbumin were purchased from Primm srl.

Techniques: Flow Cytometry, In Vitro

Journal:

Article Title: In vivo efficacy of a chitosan/IL-12 adjuvant system for protein-based vaccines

doi: 10.1016/j.biomaterials.2010.09.058

Figure Lengend Snippet: Chitosan/OVA/IL-12 stimulates IFN-γ-producing CD8+ T cells. Mice (n = 5) were vaccinated on days 0 and 14 and sacrificed on day 21. Splenocytes were pooled and cultured for 6 days with 1 μg/mL OVA257-264 (SIINFEKL) MHC class I peptide, then rested overnight in plain medium. Splenocytes (500,000 cells in 2 mL) were subsequently cultured with irradiated APCs and 1 μg/mL OVA257-264 (SIINFEKL) or control (VSV-NP) peptide for 24 h, and released IFN-γ was measured by ELISA. Results are plotted as the mean of duplicate wells.

Article Snippet: The following reagents were purchased from the suppliers indicated: ß-gal (BG13, Prozyme; Hayward, CA); FITC-, PE-, or PerCP-Cy5.5-labeled antibodies to CD8, CD3 (BD Biosciences; San Jose, CA, and eBioscience; San Diego, CA); PE-labeled MHC class I pentamers with H-2K b -restricted peptides for OVA (aa 257-264, SIINFEKL) and HSV (aa 498-505, SSIEFARL) (ProImmune; Bradenton, FL); horseradish peroxidase-conjugated goat anti-mouse IgG total, IgG1, IgG2a, and IgG2b antibodies (LifeSpan Biosciences; Seattle, WA); OVA 257-264 peptide (SIINFEKL) and VSV-NP 52-59 peptide (RGYVYQGL) (CPC Scientific; San Jose, CA).

Techniques: Cell Culture, Irradiation, Enzyme-linked Immunosorbent Assay