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. 2011 Jul 15;2(3):207–218.

Figure 3.

Figure 3.

Percentages of CD4+LLO189-201 and CD8+LLO91-99 after vaccination with ENDO/LLO. CBA/J mice vaccinated i.p with 30 µg of LLO loaded endosomes from resting BM-DM (ENDO/ LLO) for 7 days (n = 5/vaccination type). Next, mice were inoculated with LM (5 × 103 bc/mice) for 3 days. Splenocytes obtained from homogenized spleens after the vaccination protocol were stimulated for 5 hours with LLO91-99 or LLO189-201 peptides in the presence of brefeldin A for intracellular cytokine staining. LLO peptide stimulated splenocytes surface stained for CD4 or CD8 were fixed and permeabilized using a cytofix/cytoperm kit (BD. Biosciences). Stimulated cells were surface stained for CD4+ FITC-labelled or CD8+ APC-labelled and data gated to include exclusively CD4+ or CD8+ events, R2 and R3 gates, respectively (upper right flow histogram). Lower flow histograms shows the percentages of CD4+/ LLO189-201 IFN-y producers from the R2 gate (lower left) and CD8+/LLO91-99 IFN-y producers from the R3 gate (lower right), respectively. Samples are performed in triplicates and results are expressed ad the mean ± SD (p < 0.05).