Figures & data
Figure 1. IL-10R blockade enhanced CD8+ T cell responses to ChAdV63.HIVconsv in mice. (A) Gating strategy for the identification of IFN-γ+ CD8+ T cells. (B) IFN-γ and IL-2 production by HIVconsv-specific CD4+ T cells 21 d post-immunization. Mice were immunized with ChAdV63.HIVconsv in combination with anti-IL-10R (gray bars; n = 10) or isotype control antibody (white bars; n = 10). Statistical significance calculated using an unpaired t test (IFN-γ responses) or Mann-Whitney test (IL-2 responses). (C) Frequency of H-, P- and G1-specific IFN-γ+ CD8+ T cells 21 d post-immunization in mice immunized with ChAdV63.HIVconsv in combination with anti-IL-10R (gray bars; n = 10) or isotype control antibody (white bars; n = 10). (D) Total frequency of antigen-specific IFN-γ+ CD8+ T cells 21 d post-immunization in mice immunized with ChAdV63.HIVconsv in combination with anti-IL-10R (gray bar; n = 10) or isotype control antibody (white bar; n = 10). Data are representative of 2 independent experiments; statistical significance calculated using an unpaired t test. (E) Gating strategy for the identification of IFN-γ+ CD107a+ CD8+ T cells. (F) Total frequency of antigen-specific IFN-γ+ CD107a+ CD8+ T cells in mice immunized with ChAdV63.HIVconsv in combination with anti-IL-10R (gray bar; n = 10) or isotype control antibody (white bar; n = 10). Statistical significance calculated using an unpaired t test.
Figure 2. IL-10R blockade enhanced lysis of targets pulsed with a subdominant CTL epitope. (A) IFN-γ ELISPOT responses 21 d post immunization to 80 pools containing peptides spanning the HIVconsv immunogen in mice immunized with ChAdV63.HIVconsv in combination with anti-IL-10R (gray bars; n = 2) or isotype control antibody (white bars; n = 2). (B) IFN-γ ELISPOT responses to peptide 42 (left panel) and peptide 43 (right panel) in mice immunized with ChAdV63.HIVconsv in combination with anti-IL-10R (gray bars; n = 5) or isotype control antibody (white bars; n = 5). (C) Representative plot showing peptide 42-pulsed and unpulsed target cells isolated 16 hours after injection into a mouse immunized 21 d previously with ChAdV63.HIVconsv. (D) Proportion of peptide 42-pulsed targets killed in vivo in mice immunized 21 d previously with ChAdV63.HIVconsv in combination with anti-IL-10R (gray bar; n = 10) or isotype control antibody (white bar; n = 10). Statistical significance calculated using an unpaired t test.
Figure 3. The effect of IL-10R blockade on innate responses to ChAdV63.HIVconsv. (A) Representative histogram showing CD86 expression on CD11c+ cells 24 hours post-immunization in mice immunized with ChAdV63.HIVconsv in combination with either anti-IL-10R antibody (solid line) or an irrelevant isotype control antibody (dashed line). (B) Gating strategy for the identification of CD11c+ cells and representative plot showing the percentage of CD11c+ cells that are CD3- (far right plot). (C) CD86 expression on CD11c+ cells 24 hours post-immunization in mice immunized with ChAdV63.HIVconsv in combination with either anti-IL-10R antibody (gray bar; n = 5) or isotype control antibody (white bar; n = 5). Statistical significance was determined using the Mann Whitney test. (D) The proportion of splenocytes expressing CD11c 24 hours post-immunization in mice immunized with ChAdV63.HIVconsv after receiving anti-IL-10R (gray box; n = 5) or isotype control antibody (white box; n = 5). (E) Concentration of IP-10, MCP-1 and TNF-α in serum collected 24 hours post-immunization from mice immunized with ChAdV63.HIVconsv in combination with anti-IL-10R (gray bars; n = 10) or isotype control antibody (white bars; n = 10). Data are representative of 2 independent experiments.
