Figure 6.

Vector Expressing CD40L and PD-1 Microbody Provides an Accelerated and More Functional T Response

(A) BLT mice were injected with empty vector HIV.PD1-mtCD40L, HIV.CD40L, or HIV.PD1-CD40L-transduced HSC-DCs twice over a 2-week period. One week later, the mice were challenged with HIV-1. Results represent aggregate data from three independent experiments in which BLT humanized mice were generated from three different non-HLA-A*0201-restricted donors (n = 6–26/group). (B) Peripheral HIV-1 loads were determined every 2 weeks by qRT-PCR. Pooled data are shown. Line graphs (left) represent mean ± SEM. Bar graphs showing individual mice (right) represent mean ± SD. *p < 0.05, **p < 0.01 by Mann-Whitney U tests. (C) At week 8 post-HIV-1 challenge, mouse splenocytes were pulsed with a pool of clade B HIV-1 peptides, and the percentage of CD107a⁺ and IFNγ⁺ T cells was determined by intracellular staining and flow cytometry. Representative plots from one mouse per group for CD107a expression are shown (left), and pooled analyses of the mice for CD107a and IFNγ expression are shown (right) (n = 5–8/group). Data represent mean ± SD. *p < 0.05, **p < 0.01 by Mann-Whitney U tests. (D) BLT humanized mice were injected with untransduced HSC-DCs or HIV.GFP, HIV.CD40L, HIV.PD1-mtCD40L, or HIV.PD1-CD40L-transduced HSC-DCs twice over a 2-week period and 1 week later challenged with HIV-1 (n = 4–5/group). At week 4 (left) and week 6 (right) post-HIV-1 challenge, PBMCs from each group were pooled and analyzed as in (C).