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. 2011 Aug 15;187(4):1634-42.
doi: 10.4049/jimmunol.1100077. Epub 2011 Jul 8.

4-1BB signaling synergizes with programmed death ligand 1 blockade to augment CD8 T cell responses during chronic viral infection

Vaiva Vezys  1 , Pablo Penaloza-MacMaster, Daniel L Barber, Sang-Jun Ha, Bogumila Konieczny, Gordon J Freeman, Robert S Mittler, Rafi Ahmed
Affiliations

4-1BB signaling synergizes with programmed death ligand 1 blockade to augment CD8 T cell responses during chronic viral infection

Vaiva Vezys et al. J Immunol. .

Abstract

Previous studies have identified the inhibitory role that the programmed death 1 (PD-1) pathway plays during chronic infection. Blockade of this pathway results in rescue of viral-specific CD8 T cells, as well as reduction of viral loads in mice chronically infected with lymphocytic choriomeningitis virus (LCMV). We tested the effect of combining PD ligand 1 (PD-L1) blockade with an agonistic regimen that induces 4-1BB costimulation during chronic LCMV infection. There is a boosting effect in the rescue of LCMV-specific CD8 T cell responses after dual treatment with PD-L1 blockade and 4-1BB agonistic Abs when the amount and timing of 4-1BB costimulation are carefully controlled. When PD-L1-blocking Abs are given together with a single low dose of anti-4-1BB agonistic Abs, there is an enhanced and stable expansion of viral-specific CD8 T cells. Conversely, when blocking Abs to PD-L1 are given with a repetitive high dose of anti-4-1BB, there is an initial synergistic expansion of viral-specific CD8 T cells by day 7, followed by dramatic apoptosis by day 14. Viral control paralleled CD8 T cell kinetics after dual treatment. By day 7 posttreatment, viral titers were lower in both of the combined regimens (compared with PD-L1 blockade alone). However, whereas the high dose of anti-4-1BB plus PD-L1 blockade resulted in rebound of viral titers to original levels, the low dose of anti-4-1BB plus PD-L1 blockade resulted in a stable reduction of viral loads. These findings demonstrate the importance of carefully manipulating the balance between activating and inhibitory signals to enhance T cell responses during chronic infection.

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Conflict of interest statement

Disclosures

R.A., G.J.F., D.L.B., and S.-J.H. have patents and receive patent royalties related to the PD-1 pathway. The other authors have no financial conflicts of interest.

Figures

FIGURE 1
FIGURE 1
High dose of 4-1BB stimulation together with PD-L1 blockade results in a transient increase of virus-specific CD8 T cells. A, Experimental set-up. Mice received CD4-depleting Ab (GK1.5) the day before and the day of infection with LCMV clone 13. After 45 d, mice received multiple doses of anti–4-1BB agonistic Abs (200 μg) together with 200 μg PD-L1–blocking Abs every 3 d (five times). B, Percentage of splenic CD8 T cells that are specific for the LCMV immunodominant Dbgp276–286 epitope at the indicated times after start of Ab treatment. C, Total numbers of splenic CD8 T cells that are specific for the LCMV immunodominant Db gp276–286 epitope at the indicated times after start of Ab treatment. D, Serum LCMV titers. *p < 0.05. Bars indicate SEM; n = 15–21; experiments were repeated five times.
FIGURE 2
FIGURE 2
Low dose of 4-1BB stimulation together with PD-L1 blockade results in a permanent increase in virus-specific CD8 T cells. A, Experimental set-up. Mice received CD4-depleting Ab (GK1.5) the day before and the day of infection with LCMV clone 13. After 45 d, mice received a single dose of anti–4-1BB agonistic Abs (50 μg) together with 200 μg PD-L1–blocking Abs at day 1. PD-L1 blockade alone was continued every 3 d (five times). B, Percentage of splenic CD8 T cells that are specific for the LCMV immunodominant Dbgp276–286 epitope at the indicated times after start of Ab treatment. C, Total numbers of splenic CD8 T cells that are specific for the LCMV immunodominant Dbgp276–286 epitope at the indicated times after start of Ab treatment. D, Serum LCMV titers. *p < 0.05. Bars indicate SEM; n = 15–21; experiments were repeated six times.
FIGURE 3
FIGURE 3
Low dose of 4-1BB stimulation together with PD-L1 blockade increases cytokine production from LCMV-specific CD8 T cells. Experimental set-up as in Fig. 2A. A, Percentage of CD8 T cells from spleen-producing cytokines after 5 h stimulation with several LCMV peptides. B, Total numbers of IFN-γ cytokine-producing CD8 T cells after 5 h stimulation with several LCMV peptides. C, Total numbers of dual IFN-γ and TNF-α cytokine-producing CD8 T cells after 5 h stimulation with LCMV peptides. *p = 0.05, **p = 0.0016, ***p = 0.0010. Experiments were repeated four times.
FIGURE 4
FIGURE 4
Low dose of 4-1BB stimulation together with PD-L1 blockade results in marked phenotypic and functional differences in LCMV-specific CD8 T cells. Experimental set-up as in Fig. 2A. A, First row, Flow plots are gated on total lymphocytes in spleen. Bottom numbers represent percentage CD8+ T cells within the total lymphocyte population. Top numbers represent the percentage of CD8+ T cells that are activated (CD44hi) within the total lymphocyte population. Second row, Flow plots are gated on CD8 T cells. Numbers indicate the percentage of cells that are PD-1hiCD44hi within the CD8 population (representing total expanded CD8 T cells). B, Granzyme B, CD62L, Ki67, and CD127 expression on Db gp276–286-specific CD8 T cells in spleen. Plots are gated on CD8 T cells. Experiments were repeated at least three times.
FIGURE 5
FIGURE 5
LCMV-specific CD8 T cell viability after cotreatment with high or low dose of anti–4-1BB together with PD-L1 blockade. Experimental set-up as in Figs. 1A or 2A. A, FACS plot showing the percentage of splenic Dbgp276–286-specific CD8 T cells that are apoptotic or alive by annexin V/7AAD staining in spleen at days 7 and 14 posttreatment. B, Percent of Dbgp276–286-specific CD8 T cells that are positive for both annexin V and 7AAD (apoptotic population). C, Percentage of Dbgp276–286-specific CD8 T cells that are negative for both annexin V and 7AAD (42). *p < 0.05. Experiments were repeated three times.
FIGURE 6
FIGURE 6
Low dose of 4-1BB stimulation together with PD-L1 blockade during a milder chronic infection does not result in enhanced CD8 T cell rescue compared with PD-L1 blockade alone. A, Experimental set-up. Mice were infected with LCMV clone 13 (without prior CD4 depletion). Around days 15–21 postinfection, mice received a single dose of anti–4-1BB agonistic Abs (50 μg) together with PD-L1 blockade on day 1. PD-L1 blockade alone was continued every 3 d (five times). B, Percentage of CD8 T cells from spleen producing cytokines after 5 h stimulation with several LCMV peptides. C, Total numbers of cytokine producing CD8+ splenocytes after 5 h stimulation with LCMV peptides. D, LCMV serum titers. IgG treatment was similar to anti–4-1BB alone. Experiment was repeated twice; n = 12.
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References

    1. Wherry EJ, Blattman JN, Murali-Krishna K, van der Most R, Ahmed R. Viral persistence alters CD8 T-cell immunodominance and tissue distribution and results in distinct stages of functional impairment. J Virol. 2003;77:4911–4927. - PMC - PubMed
    1. Yi JS, Cox MA, Zajac AJ. T-cell exhaustion: characteristics, causes and conversion. Immunology. 2010;129:474–481. - PMC - PubMed
    1. Zajac AJ, Blattman JN, Murali-Krishna K, Sourdive DJ, Suresh M, Altman JD, Ahmed R. Viral immune evasion due to persistence of activated T cells without effector function. J Exp Med. 1998;188:2205–2213. - PMC - PubMed
    1. Barber DL, Wherry EJ, Masopust D, Zhu B, Allison JP, Sharpe AH, Freeman GJ, Ahmed R. Restoring function in exhausted CD8 T cells during chronic viral infection. Nature. 2006;439:682–687. - PubMed
    1. Day CL, Kaufmann DE, Kiepiela P, Brown JA, Moodley ES, Reddy S, Mackey EW, Miller JD, Leslie AJ, DePierres C, et al. PD-1 expression on HIV-specific T cells is associated with T-cell exhaustion and disease progression. Nature. 2006;443:350–354. - PubMed

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