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Memory programming in CD8(+) T-cell differentiation is intrinsic and is not determined by CD4 help.

Kim J, Ryu SJ, Oh K, Ju JM, Jeon JY, Nam G, Lee DS, Kim HR, Young Kim J, Chang J, Sproule T, Choi K, Roopenian D, Choi EY - Nat Commun (2015)

Bottom Line: Contrary to the general consensus that CD4 help encodes memory programmes in CD8(+) T cells and helper-deficient CD8(+) T cells are abortive, these cells can differentiate into effectors and memory precursors.Importantly, accelerating antigen clearance or simply increasing the burst effector size enables generation of memory cells by CD8(+) T cells, regardless of CD4 help.These results suggest that the memory programme is CD8(+) T-cell-intrinsic, and provide insight into the role of CD4 help in CD8(+) T-cell responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 110-799, Korea.

ABSTRACT
CD8(+) T cells activated without CD4(+) T-cell help are impaired in memory expansion. To understand the underlying cellular mechanism, here we track the dynamics of helper-deficient CD8(+) T-cell response to a minor histocompatibility antigen by phenotypic and in vivo imaging analyses. Helper-deficient CD8(+) T cells show reduced burst expansion, rapid peripheral egress, delayed antigen clearance and continuous activation, and are eventually exhausted. Contrary to the general consensus that CD4 help encodes memory programmes in CD8(+) T cells and helper-deficient CD8(+) T cells are abortive, these cells can differentiate into effectors and memory precursors. Importantly, accelerating antigen clearance or simply increasing the burst effector size enables generation of memory cells by CD8(+) T cells, regardless of CD4 help. These results suggest that the memory programme is CD8(+) T-cell-intrinsic, and provide insight into the role of CD4 help in CD8(+) T-cell responses.

No MeSH data available.


Related in: MedlinePlus

Exhaustion of H60-specific CD8+ T cells under helper-deficient conditions.(a) Flow cytometric analysis of J15 CD8+ T cells (transferred in high numbers) for expression of PD-1 and Blimp-1 on day 39 post priming (n=2 per group per experiment). (b,c) Flow cytometric analyses of (b) J15 CD8+ T cells transferred in low numbers and (c) polyclonal H60-tetramer-binding CD8+ T cells on day 39 post priming. Percentages of the positive population are plotted (n=2 per group). (d) Real-time quantitative RT–PCR analysis to determine relative expression levels of Pdcd1 (PD-1), Prdm1 (Blimp-1), Tim-3 (Tim-3), Lag3 (Lag3) and Sell (CD62L). RNA was isolated from splenic J15 CD8+ T cells in helped or helper-deficient adoptive hosts transferred in high numbers on day 39 post priming. Delta–delta Ct values were normalized to those obtained from amplification of β-actin and were expressed as fold changes compared with helped J15 gene profiles. Data represent the means±s.e.m. of two independent experiments (n=5 per group). (e) Expressions of CD44 and IFN-γ by J15 CD8+ T cells on day 2 post boosting of the adoptive hosts transferred in high numbers. Percentages of positive cells in CD45.1+J15 CD8+ T cells are plotted (n=2 per group). (f) Helper-deficient primed mice were treated with anti-PD-L1 mAb or control Rat IgG2b once every 3 days from day 14 post priming (five times). The primed-and-antibody-treated mice were boosted on day 40 post priming (n=3 per group). (g) Female PD-1-deficient or WT B6 mice were challenged according to the tolerizing and boosting schedules (n=3 per group). (f,g) Longitudinal flow cytometric analysis of PBLs was performed and flow cytometric data obtained on day 10 post priming (1o peak) and day 7 post boosting (2o peak) are presented with plots of fold change in frequencies of H60-tetramer-binding CD8+ T cells. (h) BLI analysis of antigen clearance. B6-albino hosts adoptively transferred (A.T.) with Pdcd1−/− or WT J15 CD8+ T cells were monitored periodically after 1 × or 3 × helper-deficient priming with B6.CH60-LucTg cells (n=2 per group). Data shown represent two (b–f) or at least three (a,g,h) independent experiments. Data are presented as means±s.e.m. *P<0.05, **P<0.01, ***P<0.001 determined by Student's t-test.
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f4: Exhaustion of H60-specific CD8+ T cells under helper-deficient conditions.(a) Flow cytometric analysis of J15 CD8+ T cells (transferred in high numbers) for expression of PD-1 and Blimp-1 on day 39 post priming (n=2 per group per experiment). (b,c) Flow cytometric analyses of (b) J15 CD8+ T cells transferred in low numbers and (c) polyclonal H60-tetramer-binding CD8+ T cells on day 39 post priming. Percentages of the positive population are plotted (n=2 per group). (d) Real-time quantitative RT–PCR analysis to determine relative expression levels of Pdcd1 (PD-1), Prdm1 (Blimp-1), Tim-3 (Tim-3), Lag3 (Lag3) and Sell (CD62L). RNA was isolated from splenic J15 CD8+ T cells in helped or helper-deficient adoptive hosts transferred in high numbers on day 39 post priming. Delta–delta Ct values were normalized to those obtained from amplification of β-actin and were expressed as fold changes compared with helped J15 gene profiles. Data represent the means±s.e.m. of two independent experiments (n=5 per group). (e) Expressions of CD44 and IFN-γ by J15 CD8+ T cells on day 2 post boosting of the adoptive hosts transferred in high numbers. Percentages of positive cells in CD45.1+J15 CD8+ T cells are plotted (n=2 per group). (f) Helper-deficient primed mice were treated with anti-PD-L1 mAb or control Rat IgG2b once every 3 days from day 14 post priming (five times). The primed-and-antibody-treated mice were boosted on day 40 post priming (n=3 per group). (g) Female PD-1-deficient or WT B6 mice were challenged according to the tolerizing and boosting schedules (n=3 per group). (f,g) Longitudinal flow cytometric analysis of PBLs was performed and flow cytometric data obtained on day 10 post priming (1o peak) and day 7 post boosting (2o peak) are presented with plots of fold change in frequencies of H60-tetramer-binding CD8+ T cells. (h) BLI analysis of antigen clearance. B6-albino hosts adoptively transferred (A.T.) with Pdcd1−/− or WT J15 CD8+ T cells were monitored periodically after 1 × or 3 × helper-deficient priming with B6.CH60-LucTg cells (n=2 per group). Data shown represent two (b–f) or at least three (a,g,h) independent experiments. Data are presented as means±s.e.m. *P<0.05, **P<0.01, ***P<0.001 determined by Student's t-test.

Mentions: Antigen persistence in the helper-deficient hosts (Fig. 2a) and the above Tem/Teff phenotype in helper-deficient T cells reminded us of CD8+ T-cell exhaustion in chronically infected hosts30. Thus, we assessed the expression of PD-1 surface marker and the Blimp-1 transcription factor associated with CD8+ T-cell exhaustion31. The proportions of the PD-1+ and Blimp-1hi populations in J15 CD8+ T cells on day 39 were higher in helper-deficient hosts than in helped hosts (Fig. 4a). The high proportions of PD-1+ cells in the helper-deficient T cells were verified using J15 CD8+ T cells transferred in low numbers and polyclonal H60-tetramer-binding CD8+ T cells (Fig. 4b,c and Supplementary Fig. 3). Quantitative reverse transcription–polymerase chain reaction analysis confirmed higher expression of genes (Pdcd1, Prdm1, Tim-3 and Lag3) relevant to exhaustion32 by helper-deficient CD8+ T cells, compared with helped cells (Fig. 4d). Furthermore, helper-deficient J15 CD8+ T cells downregulated IFN-γ and CD44 after boosting (Fig. 4e), another characteristic of exhausted CD8+ T cells33.


Memory programming in CD8(+) T-cell differentiation is intrinsic and is not determined by CD4 help.

Kim J, Ryu SJ, Oh K, Ju JM, Jeon JY, Nam G, Lee DS, Kim HR, Young Kim J, Chang J, Sproule T, Choi K, Roopenian D, Choi EY - Nat Commun (2015)

Exhaustion of H60-specific CD8+ T cells under helper-deficient conditions.(a) Flow cytometric analysis of J15 CD8+ T cells (transferred in high numbers) for expression of PD-1 and Blimp-1 on day 39 post priming (n=2 per group per experiment). (b,c) Flow cytometric analyses of (b) J15 CD8+ T cells transferred in low numbers and (c) polyclonal H60-tetramer-binding CD8+ T cells on day 39 post priming. Percentages of the positive population are plotted (n=2 per group). (d) Real-time quantitative RT–PCR analysis to determine relative expression levels of Pdcd1 (PD-1), Prdm1 (Blimp-1), Tim-3 (Tim-3), Lag3 (Lag3) and Sell (CD62L). RNA was isolated from splenic J15 CD8+ T cells in helped or helper-deficient adoptive hosts transferred in high numbers on day 39 post priming. Delta–delta Ct values were normalized to those obtained from amplification of β-actin and were expressed as fold changes compared with helped J15 gene profiles. Data represent the means±s.e.m. of two independent experiments (n=5 per group). (e) Expressions of CD44 and IFN-γ by J15 CD8+ T cells on day 2 post boosting of the adoptive hosts transferred in high numbers. Percentages of positive cells in CD45.1+J15 CD8+ T cells are plotted (n=2 per group). (f) Helper-deficient primed mice were treated with anti-PD-L1 mAb or control Rat IgG2b once every 3 days from day 14 post priming (five times). The primed-and-antibody-treated mice were boosted on day 40 post priming (n=3 per group). (g) Female PD-1-deficient or WT B6 mice were challenged according to the tolerizing and boosting schedules (n=3 per group). (f,g) Longitudinal flow cytometric analysis of PBLs was performed and flow cytometric data obtained on day 10 post priming (1o peak) and day 7 post boosting (2o peak) are presented with plots of fold change in frequencies of H60-tetramer-binding CD8+ T cells. (h) BLI analysis of antigen clearance. B6-albino hosts adoptively transferred (A.T.) with Pdcd1−/− or WT J15 CD8+ T cells were monitored periodically after 1 × or 3 × helper-deficient priming with B6.CH60-LucTg cells (n=2 per group). Data shown represent two (b–f) or at least three (a,g,h) independent experiments. Data are presented as means±s.e.m. *P<0.05, **P<0.01, ***P<0.001 determined by Student's t-test.
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Related In: Results  -  Collection

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f4: Exhaustion of H60-specific CD8+ T cells under helper-deficient conditions.(a) Flow cytometric analysis of J15 CD8+ T cells (transferred in high numbers) for expression of PD-1 and Blimp-1 on day 39 post priming (n=2 per group per experiment). (b,c) Flow cytometric analyses of (b) J15 CD8+ T cells transferred in low numbers and (c) polyclonal H60-tetramer-binding CD8+ T cells on day 39 post priming. Percentages of the positive population are plotted (n=2 per group). (d) Real-time quantitative RT–PCR analysis to determine relative expression levels of Pdcd1 (PD-1), Prdm1 (Blimp-1), Tim-3 (Tim-3), Lag3 (Lag3) and Sell (CD62L). RNA was isolated from splenic J15 CD8+ T cells in helped or helper-deficient adoptive hosts transferred in high numbers on day 39 post priming. Delta–delta Ct values were normalized to those obtained from amplification of β-actin and were expressed as fold changes compared with helped J15 gene profiles. Data represent the means±s.e.m. of two independent experiments (n=5 per group). (e) Expressions of CD44 and IFN-γ by J15 CD8+ T cells on day 2 post boosting of the adoptive hosts transferred in high numbers. Percentages of positive cells in CD45.1+J15 CD8+ T cells are plotted (n=2 per group). (f) Helper-deficient primed mice were treated with anti-PD-L1 mAb or control Rat IgG2b once every 3 days from day 14 post priming (five times). The primed-and-antibody-treated mice were boosted on day 40 post priming (n=3 per group). (g) Female PD-1-deficient or WT B6 mice were challenged according to the tolerizing and boosting schedules (n=3 per group). (f,g) Longitudinal flow cytometric analysis of PBLs was performed and flow cytometric data obtained on day 10 post priming (1o peak) and day 7 post boosting (2o peak) are presented with plots of fold change in frequencies of H60-tetramer-binding CD8+ T cells. (h) BLI analysis of antigen clearance. B6-albino hosts adoptively transferred (A.T.) with Pdcd1−/− or WT J15 CD8+ T cells were monitored periodically after 1 × or 3 × helper-deficient priming with B6.CH60-LucTg cells (n=2 per group). Data shown represent two (b–f) or at least three (a,g,h) independent experiments. Data are presented as means±s.e.m. *P<0.05, **P<0.01, ***P<0.001 determined by Student's t-test.
Mentions: Antigen persistence in the helper-deficient hosts (Fig. 2a) and the above Tem/Teff phenotype in helper-deficient T cells reminded us of CD8+ T-cell exhaustion in chronically infected hosts30. Thus, we assessed the expression of PD-1 surface marker and the Blimp-1 transcription factor associated with CD8+ T-cell exhaustion31. The proportions of the PD-1+ and Blimp-1hi populations in J15 CD8+ T cells on day 39 were higher in helper-deficient hosts than in helped hosts (Fig. 4a). The high proportions of PD-1+ cells in the helper-deficient T cells were verified using J15 CD8+ T cells transferred in low numbers and polyclonal H60-tetramer-binding CD8+ T cells (Fig. 4b,c and Supplementary Fig. 3). Quantitative reverse transcription–polymerase chain reaction analysis confirmed higher expression of genes (Pdcd1, Prdm1, Tim-3 and Lag3) relevant to exhaustion32 by helper-deficient CD8+ T cells, compared with helped cells (Fig. 4d). Furthermore, helper-deficient J15 CD8+ T cells downregulated IFN-γ and CD44 after boosting (Fig. 4e), another characteristic of exhausted CD8+ T cells33.

Bottom Line: Contrary to the general consensus that CD4 help encodes memory programmes in CD8(+) T cells and helper-deficient CD8(+) T cells are abortive, these cells can differentiate into effectors and memory precursors.Importantly, accelerating antigen clearance or simply increasing the burst effector size enables generation of memory cells by CD8(+) T cells, regardless of CD4 help.These results suggest that the memory programme is CD8(+) T-cell-intrinsic, and provide insight into the role of CD4 help in CD8(+) T-cell responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 110-799, Korea.

ABSTRACT
CD8(+) T cells activated without CD4(+) T-cell help are impaired in memory expansion. To understand the underlying cellular mechanism, here we track the dynamics of helper-deficient CD8(+) T-cell response to a minor histocompatibility antigen by phenotypic and in vivo imaging analyses. Helper-deficient CD8(+) T cells show reduced burst expansion, rapid peripheral egress, delayed antigen clearance and continuous activation, and are eventually exhausted. Contrary to the general consensus that CD4 help encodes memory programmes in CD8(+) T cells and helper-deficient CD8(+) T cells are abortive, these cells can differentiate into effectors and memory precursors. Importantly, accelerating antigen clearance or simply increasing the burst effector size enables generation of memory cells by CD8(+) T cells, regardless of CD4 help. These results suggest that the memory programme is CD8(+) T-cell-intrinsic, and provide insight into the role of CD4 help in CD8(+) T-cell responses.

No MeSH data available.


Related in: MedlinePlus