Figure 6

T cell responses to pentamer-deficient TR3 vectors in RM. (A) In vitro growth curve of TR3 and TR3ΔUL128-131gag. MRC-5 cells were inoculated with TR3 or TR3ΔUL128-131gag at a MOI = 0.01 and cell culture supernatants were collected at the indicated times. Virus was titered by plaque assay on MRC-5 cells. The average titer of triplicate experiments (+/− SD) is shown. (B) UL128-131-deleted TR3 does not elicit T cell responses in RM. Two RM were inoculated with 5 × 10⁶ FFU TR3ΔUL128-131gag, one RM was inoculated with 5 × 10⁶ FFU TR3ΔUL78gag and the T cell response to HCMV-lysate or HIVgag was measured at the indicated days. (C) In vitro growth curve of PC-deficient TR3. MRC-5 cells (top) or ARPE cells (bottom) were inoculated with the indicated viruses at MOI = 0.01 (MRC-5) or MOI = 5 (ARPE) and cell culture supernatants were collected at the indicated times. Virus was titered as above and the average titer of triplicate experiments (+/− SD) is shown. (D) HIVgag-specific T cell response elicited by PC-deficient TR3. RM (n = 2 per recombinant) were inoculated with 5 × 10⁶ FFU of the indicated PC-deficient vectors on day 0 and the HIVgag-specific or HCMV-lysate CD4+ and CD8+ memory T cell response frequencies were determined in PBMC by intracellular cytokine staining for TNFα and/or IFNγ using overlapping HIVgag peptide pools at the indicated days. (E) Frequency of memory populations within the HIVgag-specific CD8⁺ memory T cells in peripheral blood of the six RM in (D) inoculated with the indicated viruses. Memory differentiation state was based on CD28 vs. CCR7 expression, delineating central memory (+/+T_CM), transitional effector memory (+/− T_TrEM), and effector memory (−/− T_EM), as designated.