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Genetic absence of PD-1 promotes accumulation of terminally differentiated exhausted CD8+ T cells.

Odorizzi PM, Pauken KE, Paley MA, Sharpe A, Wherry EJ - J. Exp. Med. (2015)

Bottom Line: Increased proliferation between days 8 and 14 postinfection is associated with subsequent decreased CD8(+) T cell survival and disruption of a critical proliferative hierarchy necessary to maintain exhausted populations long term.Ultimately, the absence of PD-1 leads to the accumulation of more cytotoxic, but terminally differentiated, CD8(+) TEX cells.They also highlight a novel role for PD-1 in preserving TEX cell populations from overstimulation, excessive proliferation, and terminal differentiation.

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Affiliation: Department of Microbiology and Penn Institute for Immunology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104 Department of Microbiology and Penn Institute for Immunology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104.

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Altered dynamics of TEX cell subsets in the absence of PD-1. WT and PD-1 KO P14 cells were mixed at a 1:1 ratio (250 cells each), adoptively transferred into naive recipient mice, and infected with LCMV clone 13. P14 responses were then analyzed at the indicated time points. (A) Summary of the MFI of Tbet and Eomes in WT and PD-1 KO P14 cells in the spleen at days 0 (naive), 8, and 15 p.i. with LCMV clone 13. (B) Representative FACS histograms of Tbet and Eomes expression in WT and PD-1 KO P14 cells for individual (left) and multiple (right) mice at day 42 p.i. in the spleen. (C) Protein expression of Tbet versus Eomes in WT and PD-1 KO P14 cells at day 42 p.i. in the spleen of individual mice. Values indicate the frequency of P14 cells that are TbethiEomeslo or EomeshiTbetlo. (D and E) Total frequency (D) and numbers (E) of WT and PD-1 KO P14 TbethiEomeslo and EomeshiTbetlo cells at day 42 p.i. in the spleen. (F) Frequency of P14 cells as a percentage of total CD8+ T cells in multiple organs at day 42 p.i. with LCMV clone 13, as indicated. (G) Cytotoxicity of sorted WT and PD-1 KO P14 cells on day 22 p.i. at an E/T ratio of 4:1 after 18-d incubation. (H) Expression of Granzyme B in naive CD8+ T cells, WT P14 cells, and PD-1 KO P14 cells in individual (left) and multiple mice (right). All error bars indicate ±SEM. *, P < 0.05; **, P < 0.01 (paired Student’s t test for A–F and H and unpaired Student’s t test for G) for all graphs (A–H). All data are representative of two to five independent experiments with at least five mice per group.
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fig6: Altered dynamics of TEX cell subsets in the absence of PD-1. WT and PD-1 KO P14 cells were mixed at a 1:1 ratio (250 cells each), adoptively transferred into naive recipient mice, and infected with LCMV clone 13. P14 responses were then analyzed at the indicated time points. (A) Summary of the MFI of Tbet and Eomes in WT and PD-1 KO P14 cells in the spleen at days 0 (naive), 8, and 15 p.i. with LCMV clone 13. (B) Representative FACS histograms of Tbet and Eomes expression in WT and PD-1 KO P14 cells for individual (left) and multiple (right) mice at day 42 p.i. in the spleen. (C) Protein expression of Tbet versus Eomes in WT and PD-1 KO P14 cells at day 42 p.i. in the spleen of individual mice. Values indicate the frequency of P14 cells that are TbethiEomeslo or EomeshiTbetlo. (D and E) Total frequency (D) and numbers (E) of WT and PD-1 KO P14 TbethiEomeslo and EomeshiTbetlo cells at day 42 p.i. in the spleen. (F) Frequency of P14 cells as a percentage of total CD8+ T cells in multiple organs at day 42 p.i. with LCMV clone 13, as indicated. (G) Cytotoxicity of sorted WT and PD-1 KO P14 cells on day 22 p.i. at an E/T ratio of 4:1 after 18-d incubation. (H) Expression of Granzyme B in naive CD8+ T cells, WT P14 cells, and PD-1 KO P14 cells in individual (left) and multiple mice (right). All error bars indicate ±SEM. *, P < 0.05; **, P < 0.01 (paired Student’s t test for A–F and H and unpaired Student’s t test for G) for all graphs (A–H). All data are representative of two to five independent experiments with at least five mice per group.

Mentions: Recent work has demonstrated that two subsets of CD8+ TEX cells, Tbethi progenitors and Eomeshi progeny, exist in a proliferative hierarchy to sustain long-term CD8+ T cell responses during chronic viral infections (Paley et al., 2012). Based on these observations, we next investigated whether loss of PD-1 impacted the balance between these two subsets and whether changes in these subsets could account for the altered TCR responsiveness, proliferation, and stability of the PD-1 KO P14 population. To test this idea, we assessed expression of Tbet and Eomes at days 8, 15, and 42 p.i. in WT and PD-1 KO P14 cells. At day 8 p.i., increased expression of both Tbet and Eomes was observed in PD-1 KO P14 cells, which is consistent with the increased activation of these cells early in infection (Fig. 6 A). However, on day 15 p.i., PD-1 KO P14 cells had a significant decrease in Tbet expression (Fig. 6 A), which was followed by a substantial reduction in Tbet MFI on day 42 p.i. (Fig. 6 B). Reduced Tbet expression on a per-cell basis in PD-1 KO P14 cells was accompanied by a significant decline in the frequency and total number of Tbethi progenitor cells compared with WT P14 cells (Fig. 6, C–E). As a result, the PD-1 KO P14 cell population was skewed toward the Eomeshi subset, as demonstrated by an increase in the frequency of Eomeshi cells (Fig. 6, C and D). There was also a trend toward increased total numbers of Eomeshi cells in the PD-1 KO P14 cell population (Fig. 6 E). In addition, PD-1 KO P14 cells had increased Eomes expression on a per-cell basis at days 15 (Fig. 6 A) and 42 p.i. (Fig. 6 B).


Genetic absence of PD-1 promotes accumulation of terminally differentiated exhausted CD8+ T cells.

Odorizzi PM, Pauken KE, Paley MA, Sharpe A, Wherry EJ - J. Exp. Med. (2015)

Altered dynamics of TEX cell subsets in the absence of PD-1. WT and PD-1 KO P14 cells were mixed at a 1:1 ratio (250 cells each), adoptively transferred into naive recipient mice, and infected with LCMV clone 13. P14 responses were then analyzed at the indicated time points. (A) Summary of the MFI of Tbet and Eomes in WT and PD-1 KO P14 cells in the spleen at days 0 (naive), 8, and 15 p.i. with LCMV clone 13. (B) Representative FACS histograms of Tbet and Eomes expression in WT and PD-1 KO P14 cells for individual (left) and multiple (right) mice at day 42 p.i. in the spleen. (C) Protein expression of Tbet versus Eomes in WT and PD-1 KO P14 cells at day 42 p.i. in the spleen of individual mice. Values indicate the frequency of P14 cells that are TbethiEomeslo or EomeshiTbetlo. (D and E) Total frequency (D) and numbers (E) of WT and PD-1 KO P14 TbethiEomeslo and EomeshiTbetlo cells at day 42 p.i. in the spleen. (F) Frequency of P14 cells as a percentage of total CD8+ T cells in multiple organs at day 42 p.i. with LCMV clone 13, as indicated. (G) Cytotoxicity of sorted WT and PD-1 KO P14 cells on day 22 p.i. at an E/T ratio of 4:1 after 18-d incubation. (H) Expression of Granzyme B in naive CD8+ T cells, WT P14 cells, and PD-1 KO P14 cells in individual (left) and multiple mice (right). All error bars indicate ±SEM. *, P < 0.05; **, P < 0.01 (paired Student’s t test for A–F and H and unpaired Student’s t test for G) for all graphs (A–H). All data are representative of two to five independent experiments with at least five mice per group.
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fig6: Altered dynamics of TEX cell subsets in the absence of PD-1. WT and PD-1 KO P14 cells were mixed at a 1:1 ratio (250 cells each), adoptively transferred into naive recipient mice, and infected with LCMV clone 13. P14 responses were then analyzed at the indicated time points. (A) Summary of the MFI of Tbet and Eomes in WT and PD-1 KO P14 cells in the spleen at days 0 (naive), 8, and 15 p.i. with LCMV clone 13. (B) Representative FACS histograms of Tbet and Eomes expression in WT and PD-1 KO P14 cells for individual (left) and multiple (right) mice at day 42 p.i. in the spleen. (C) Protein expression of Tbet versus Eomes in WT and PD-1 KO P14 cells at day 42 p.i. in the spleen of individual mice. Values indicate the frequency of P14 cells that are TbethiEomeslo or EomeshiTbetlo. (D and E) Total frequency (D) and numbers (E) of WT and PD-1 KO P14 TbethiEomeslo and EomeshiTbetlo cells at day 42 p.i. in the spleen. (F) Frequency of P14 cells as a percentage of total CD8+ T cells in multiple organs at day 42 p.i. with LCMV clone 13, as indicated. (G) Cytotoxicity of sorted WT and PD-1 KO P14 cells on day 22 p.i. at an E/T ratio of 4:1 after 18-d incubation. (H) Expression of Granzyme B in naive CD8+ T cells, WT P14 cells, and PD-1 KO P14 cells in individual (left) and multiple mice (right). All error bars indicate ±SEM. *, P < 0.05; **, P < 0.01 (paired Student’s t test for A–F and H and unpaired Student’s t test for G) for all graphs (A–H). All data are representative of two to five independent experiments with at least five mice per group.
Mentions: Recent work has demonstrated that two subsets of CD8+ TEX cells, Tbethi progenitors and Eomeshi progeny, exist in a proliferative hierarchy to sustain long-term CD8+ T cell responses during chronic viral infections (Paley et al., 2012). Based on these observations, we next investigated whether loss of PD-1 impacted the balance between these two subsets and whether changes in these subsets could account for the altered TCR responsiveness, proliferation, and stability of the PD-1 KO P14 population. To test this idea, we assessed expression of Tbet and Eomes at days 8, 15, and 42 p.i. in WT and PD-1 KO P14 cells. At day 8 p.i., increased expression of both Tbet and Eomes was observed in PD-1 KO P14 cells, which is consistent with the increased activation of these cells early in infection (Fig. 6 A). However, on day 15 p.i., PD-1 KO P14 cells had a significant decrease in Tbet expression (Fig. 6 A), which was followed by a substantial reduction in Tbet MFI on day 42 p.i. (Fig. 6 B). Reduced Tbet expression on a per-cell basis in PD-1 KO P14 cells was accompanied by a significant decline in the frequency and total number of Tbethi progenitor cells compared with WT P14 cells (Fig. 6, C–E). As a result, the PD-1 KO P14 cell population was skewed toward the Eomeshi subset, as demonstrated by an increase in the frequency of Eomeshi cells (Fig. 6, C and D). There was also a trend toward increased total numbers of Eomeshi cells in the PD-1 KO P14 cell population (Fig. 6 E). In addition, PD-1 KO P14 cells had increased Eomes expression on a per-cell basis at days 15 (Fig. 6 A) and 42 p.i. (Fig. 6 B).

Bottom Line: Increased proliferation between days 8 and 14 postinfection is associated with subsequent decreased CD8(+) T cell survival and disruption of a critical proliferative hierarchy necessary to maintain exhausted populations long term.Ultimately, the absence of PD-1 leads to the accumulation of more cytotoxic, but terminally differentiated, CD8(+) TEX cells.They also highlight a novel role for PD-1 in preserving TEX cell populations from overstimulation, excessive proliferation, and terminal differentiation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology and Penn Institute for Immunology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104 Department of Microbiology and Penn Institute for Immunology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104.

Show MeSH
Related in: MedlinePlus