Article Figures & Data
Figures
- Fig 1
Clinical and virological outcome of vaccinations. (A) Schematic representation of the immunization regimens. The arrows indicate times of immunizations and of challenge exposure to SIVmac251, performed by the intrarectal route at week 74 in all animals and at week 52 in the 1 × MVA group. (B, C, D, E) Plasma virus levels are presented for each animal. *, identified Mamu-A*01+ macaques. (F, G, H) Comparison of mean virus load in plasma in macaques from the different groups.
- Fig 2
Antibody titers and kinetics of CD8+CM9+ T-cell response in the vaccinated macaques. Endpoint antibody titers to p27 Gag (A) and gp120 (B) in the sera of the immunized monkeys. (C) Correlative analysis between the titers of the gp120-binding antibodies in all the vaccinated animals and plasma viral loads at week 6 postinfection. (D to G) Frequency and kinetics of CM9+CD8+ T cells in blood of the macaques during immunization and following challenge exposure to SIVmac251. (H to K) Relative frequency of CD95+CD28− (effector memory [EM] in solid black bars) and CD95+CD28+ (central memory [CM] in white bars) CM9+CD8+ T cells in blood of vaccinated and control macaques following challenge exposure to SIVmac251. The data are shown as means for each group, calculated using the individual frequency of EM or CM on the total Gag-CM9+ memory population.
- Fig 3
PD-1 expression on CD8+CM9+ T-cell response in the vaccinated macaques. (A) CFSE gating strategy and mean fluorescence intensity (MFI) of PD-1 expression on nonproliferating CFSEhi CM9+CD8+ T cells from representative macaques. (B to D) PD-1 level of expression on CFSElo or CFSEhi CM9+ CD8+ T cells from animals from the 2 × MVA and 1 × MVA/Ad5 groups were analyzed at week 74 (before challenge) and at weeks 7 and 24 after challenge exposure to SIVmac251. (E) PD-1 expression levels on low and high proliferating CD8+CM9+ T cells in the two groups of vaccinated animals at all time points. The plot includes all the data points, and each dot represents one macaque. (F and G) Frequency of Gag-specific T cells 5 days following stimulation with Gag peptides in the presence (or absence) of anti-PD-L1 blocking antibody; CD8+ T cells (F) and CD4+ T cells (G) are gated on CFSElow cells. (H to J) PD-L1 expression on human mDCs (H) and on pDCs (J) and CD86 expression on human mDCs (I) following in vitro infection with Ad5 or MVA.
- Fig 4
Heat map representation of gene expression within CM9+ CD8+ T cells at 24 weeks following challenge. (Upper panel) Genes from representative pathways analyzed with GSEA and displayed as a heat map to demonstrate differential expression between Adv CM9+ and MVA CM9+ cells. Each pathway comprising selected top upregulated genes is represented by a different color. The false color expression in log2 scale is depicted on the right side of the figure. The heat map shows the expression level of each gene (red, upregulated; blue, downregulated). Eight significant pathways are shown (P values < 0.05) (6 upregulated in Ad5 CM9+ samples and 2 upregulated in MVA CM9+ cells), as is a subset of 30 most up- and downregulated genes (22 upregulated in Ad5 CM9+ samples and 8 upregulated in MVA CM9+ samples). (Lower panel) Genes upregulated and downregulated in antigen presentation pathways.
Tables
- Table 1
Definition of pathways and genes differentially expressed in Adv versus MVA Tet+ cells
Pathway or gene name Function or description Pathways HSA04010_MAPK_SIGNALING_PATHWAY Genes involved in MAPK signaling pathway HSA04540_GAP_JUNCTION Genes involved in gap junction M_2.9_ERK_TRANSACTIVATION_CYTOSKELETAL_MAPK_JNK Progressively downregulated through 12 h following treatment of WS1 human skin fibroblasts with UVC at a high dose UVC_HIGH_D4_DN Downregulated at any time point following treatment of XPB/CS fibroblasts with 3 J/m2 UVC UVC_XPCS_ALL_DN Downregulated at 4 h following treatment of XPB/CS fibroblasts with 3 J/m2 UVC (genes involved in apoptosis) UVC_XPCS_4HR_DN Upregulated in mouse hematopoietic stem cells HSC_HSCANDPROGENITORS_FETAL Genes involved in antigen processing and presentation HSA04612_ANTIGEN_PROCESSING_AND_PRESENTATION Genes involved in MAPK signaling pathway Genes Upregulated in Tet+ MVA compared to Tet+ Adv TAPBP Required for MHC formation CIITA Involved in transcription of HLA class II and important for antigen presentation HLA-DQB1 Involved in transcription of HLA class II and important for antigen presentation HLA-DMB Helps to load HLA class II IRAK1 IGBP1 B-cell activation PSMC5 Important for MHC-I processing CCND2 Important for cell division Uppregulated in Tet+ Adv compared to Tet+ MVA MAP3K4 Apoptosis JUN Apoptosis ATF4 Apoptosis ATF2 Apoptosis RAP1A Plays a role in apoptosis DUSP5 Apoptosis TAOK1 Apoptosis PTEN Tumor suppressor ITPR1 Apoptotic gene TUBB2B PRKACB Apoptosis PRKCB1 Apoptotic gene SOS1 Differential role UBE2G1 Ubiquitination POGZ Gene suppressor ARID5B Gene repression PPP2R5C Negative control of cell division TRIM33 Gene repression SPEN Gene transcription repression CDYL Oncogene repression CNOT4 Transcription repression CBLB Anti-inflammatory
Supplemental material
Files in this Data Supplement:
- Supplemental file 1 -
Fig. S1 (Proliferative responses to the SIVmac251 gp120 Env and p27 Gag proteins before SIV challenge in the 2xMVA and 1xMVA/Ad5 groups; γ-ELISPOT responses measured in blood using the peptide pool derived from the entire SIVmac239 Gag, Env, and Nef proteins; intracellular cytokine production (IFN-γ or IL-2) in PBMCs at 2 weeks following the last immunization with MVA or Ad5 in CD8+ and CD4+ T-cells; plasma virus levels presented for each animal in Mamu-A*01+ macaques for each group.)
Fig. S2 (Upregulation and downregulation of genes involved in T-cell signaling in NFAT and CD28 cells.)
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- Supplemental file 1 -
