Oral vaccination with Liporale™ BCG induces effector and central memory Ag85B-specific CD4⁺ T cells in the spleen.

<p>Lymphocytes from the spleens of naïve, Liporale™-BCG vaccinated (BCG oral) or subcutaneously vaccinated (BCG s.c.) mice were stained with an Ag85B/MHCII tetramer and enriched for tetramer positive cells by magnetic bead isolation. (A) Representative flow cytometry density plots showing CD62L and CD44 expression on total CD4⁺ T cells from spleens of naïve mice, or Ag85b-specific CD4⁺ T cells from the spleens of BCG vaccinated mice. (B) Bar graphs showing the proportion of naïve (CD62L^hi, CD44^lo), T_EFF/T_EM (CD62L^lo, CD44^hi) or T_CM (CD62L^hi, CD44^hi) CD4⁺ T lymphocytes of total CD4⁺ T cells from naïve mice or Ag85B-specific CD4⁺ T cells from the spleens of BCG vaccinated mice at 4, 8 and 30 weeks post vaccination. Results are displayed as mean + SEM of n = 5 for each group: *p<0.05, **p<0.01, ***p<0.001 (Mann-Whitney test). Eight and 30 weeks results are representative of 2 independent experiments.</p

Oral vaccination with Liporale™-BCG induces CD4⁺ T cell cytokine production in the lung.

<p>Lymphocytes from the lungs of naïve, Liporale™-BCG vaccinated (BCG oral) or subcutaneous vaccinated (BCG s.c.) mice were stimulated for 6 hours <i>in vitro</i> in the presence of Brefeldin A and monensin then analyzed by flow cytometry. (A) Representative plots show CD4⁺ T cells from the lungs of naïve or BCG vaccinated mice expressing IFNγ, TNFα or IL-2. (B) Bar graphs show the percentage of CD4⁺ T cells from the lungs of naïve or BCG vaccinated mice expressing cytokines at 4, 8 or 30 weeks post immunization. Results are displayed as mean + SEM of n = 5 for each group, significance expressed relative to naïve: *p<0.05, **p<0.01, ***p<0.001 (one way ANOVA with Tukey post test). Eight and 30 weeks results are representative of 2 independent experiments.</p

Oral vaccination with Liporale™-BCG increases the number of Ag85B-specific CD4⁺ T cells in the spleen.

<p>Lymphocytes from the spleens of naïve, Liporale™-BCG vaccinated (BCG oral) or subcutaneously vaccinated (BCG s.c.) mice were stained with an Ag85B/MHCII tetramer and enriched for tetramer positive cells by magnetic bead isolation. (A) Representative flow cytometry plots show Ag85B-specific CD4⁺ T cells in the spleens of naïve or BCG vaccinated mice at 4, 8 and 30 weeks post immunization. (B) Bar graphs show the number of Ag85B-specific CD4⁺ T cells in the spleens of naïve or BCG vaccinated mice at 4, 8 and 30 weeks post vaccination. Results are displayed as mean +SEM of n = 5 for each group, significance expressed relative to naïve: *p<0.05, **p<0.01, ***p<0.001 (one way ANOVA with Tukey post test). The 8 and 30 weeks results are representative of 2 independent experiments.</p

Oral vaccination with Liporale™-BCG induces long-lived CD4⁺ effector T cells in the lung.

<p>Lymphocytes from the lungs of naïve, Liporale™-BCG vaccinated (BCG oral) or subcutaneous vaccinated (BCG s.c.) mice were analyzed by flow cytometry. (A) Representative flow cytometry plots show the gating strategy used to identify CD4⁺ T cells. (B) Bar graphs show the proportion of naïve (CD62L^hi, CD44^lo), T_EFF/T_EM (CD62L^lo, CD44^hi) or T_CM (CD62L^hi, CD44^hi) CD4⁺ T lymphocytes of total CD4⁺ T cells from the lungs of naïve or BCG vaccinated mice at 4, 8 and 30 weeks post vaccination. Results are displayed as mean + SEM of n = 5 for each group, significance expressed relative to naïve: *p<0.05, **p<0.01, ***p<0.001 (one way ANOVA with Tukey post test). Eight and 30 weeks results are representative of 2 independent experiments.</p