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. 2011 Jul 15;2(3):207–218.

Table 2.

Specific LLO immune response in spleens of vaccinated mice.

Peptide LLO91-99 CD4+ T cellsb CD8+ T cells CD4+c-IFNintracell CD8+-IFNintracell
NVa 25 ± 0.02 18 ± 0.01 0.03 ± 0.001 0.1 ± 0.001
ENDO/LLO 33 ± 0.02 22 ± 0.03 0.02 ± 0.002 0.47 ± 0.002
ENDO-IFN/LLO 29 ± 0.01 23 ± 0.01 0.05 ± 0.002 0.71 ± 0.004
BM-DM/LM 35 ± 0.03 25 ± 0.02 0.04 ± 0.001 0.99 ± 0.007
BM-DM-IFN/LM 30 ± 0.02 26 ± 0.03 0.02 ± 0.002 1.4 ± 0.02
Peptide LLO189-201
NV 28 ± 0.01 19 ± 0.02 0.19 ± 0.005 0.01 ± 0.0
ENDO/LLO 31 ± 0.03 21 ± 0.03 0.40 ± 0.02 0.03 ± 0.001
ENDO-IFN/LLO 29 ± 0.02 23 ± 0.03 0.56 ± 0.04 0.03 ± 0.002
BM-DM/LM 34 ± 0.01 27 ± 0.02 0.68 ± 0.06 0.01 ± 0.001
BM-DM-IFN/LM 32 ± 0.03 28 ± 0.03 0.78 ± 0.05 0.0 ± 0.0
a

CBA/J mice were vaccinated i.p or not (NV) with 30 µg of LLO loaded endosomes from resting BM-DM (ENDO/LLO), LLO loaded endosomes from IFN-γ activated BM-DM (ENDO-IFN/LLO), LM infected resting BM-DM (1 × 106 cells) or LM infected and IFN-γ activated BM-DM (1 × 106 cells) for 7 days (n = 5/vaccination type). Next, mice were inoculated with LM (5 × 103 bc/mice) for 3 days. Splenocytes were obtained from homogenized spleens after the vaccination protocol. Samples were performed in triplicates and results expressed the mean ± SD (p < 0.01).

b

Splenocytes from vaccinated mice were stimulated for 5 hours with LLO91-99 or LLO189-201 peptides in the presence of brefeldin A for intracellular cytokine staining. Stimulated cells were surface stained for CD4 FITC-labelled or CD8 FITC-labelled. Percentages of CD4+ or CD8+ T cells are expressed for each vaccination protocol. Samples were done in triplicates and results expressed the mean ± SD (p < 0.05).

c

LLO peptide stimulated splenocytes surface stained for CD4 or CD8 were fixed and permeabilized using a cytofix/cytoperm kit (BD. Biosciences). Cells were stained for IFN-γ using an anti-IFN-γ antibody PE-labelled. Samples were acquired using a FACSCanto flow cytometer. Data were gated to include exclusively CD4+ or CD8+ events, and the percentage of these cells expressing IFN-γ was determined. Samples were done in triplicates and results expressed the mean ± SD (p < 0.05).