Comparison of response to <i>Staphylococcus</i> enterotoxin B (SEB) in samples with (Treg+) and without Treg (Treg−).

<p>Permutation comparison test showed no difference in the proportion of polyfunctional CD8⁺ T cells after Treg depletion. (P = 0.762; N = 7)</p

Gating of Treg in representative subjects.

<p>Shown above is the gating used to determined the Treg frequency in a representative normal control (A) and an HIV-1-infected subject, pre-vaccine (B). The Treg population was defined as CD3⁺CD4⁺CD25^HIFOXP3⁺.</p

Polyfunctionality of the total vaccine response with (Treg+) and without Treg (Treg−).

<p>The total vaccine response was obtained by subtracting the pre-vaccine response from the post-vaccine response and represents the vaccine-induced change in CD8⁺ T cell secretion of immune mediators from baseline. Analysis by permutation comparison showed a significant increase in polyfunctional response to Gag peptide in the samples depleted of Treg. (P = 0.029; N = 7)</p

Percentages of CD8⁺ T cells with poly vs monofunctional responses before and after vaccination.

<p>Bars are the mean percentages (+SE) of CD8⁺ T cells in paired pre- and post-vaccine samples secreting one (monofxn) or more than one (polyfxn) immune mediators (IFNγ, TNFα, IL-2, MIP-1β, and CD107a) in response to Gag peptide. (P = NS; N = 7)</p

Polyfunctional response of CD8⁺ T cells pre- and post-vaccine after Treg depletion.

<p>The response patterns are color-coded by the number of positive functions. The x-axis contains the combinations of positive immune mediators whereas the y-axis is the percentage of CD8⁺ T cells secreting each combination. The pie charts show the proportion of CD8⁺ T cells that are polyfunctional, i.e., secreting more than 1 immune mediator in response to stimulation with Gag peptide. There was increased polyfunctionality after the Treg were removed. Despite the apparent increase in polyfunctionality, the permutation comparison test showed no statistical difference between the two pie charts. (P = 0.276; N = 7)</p

Comparison of CD45RO, FOXP3, GITR, and CTLA4 expression.

<p>The mean percentage expression in CD4⁺CD25^HI T cells was compared among normal control (NC; N = 10) and the HIV-1-infected subjects pre- and post-vaccine (N = 17). CD4⁺CD25^HI expression of GITR and CTLA4 in HIV-1-infected subjects (pre- and post-vaccine) was higher than in NC. (P<0.01)</p

Treg frequency in paired pre- and post-vaccine of the 17 subjects.

<p>Percentages of CD4⁺CD25^HIFOXP3⁺ T cells before (pre-vaccine) and 2 weeks after therapeutic vaccination in each of the 17 HIV-1-infected subjects are shown. Asterisks indicate subjects who had a positive vaccine response by ELISPOT analyses. There was a trend of increased Treg frequency post-vaccine. (P = 0.06)</p