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Immune signatures of protective spleen memory CD8 T cells

View Article: PubMed Central - PubMed

ABSTRACT

Memory CD8 T lymphocyte populations are remarkably heterogeneous and differ in their ability to protect the host. In order to identify the whole range of qualities uniquely associated with protective memory cells we compared the gene expression signatures of two qualities of memory CD8 T cells sharing the same antigenic-specificity: protective (Influenza-induced, Flu-TM) and non-protective (peptide-induced, TIM) spleen memory CD8 T cells. Although Flu-TM and TIM express classical phenotypic memory markers and are polyfunctional, only Flu-TM protects against a lethal viral challenge. Protective memory CD8 T cells express a unique set of genes involved in migration and survival that correlate with their unique capacity to rapidly migrate within the infected lung parenchyma in response to influenza infection. We also enlighten a new set of poised genes expressed by protective cells that is strongly enriched in cytokines and chemokines such as Ccl1, Ccl9 and Gm-csf. CCL1 and GM-CSF genes are also poised in human memory CD8 T cells. These immune signatures are also induced by two other pathogens (vaccinia virus and Listeria monocytogenes). The immune signatures associated with immune protection were identified on circulating cells, i.e. those that are easily accessible for immuno-monitoring and could help predict vaccines efficacy.

No MeSH data available.


Comparison of phenotype and protective capacity between two memory CD8 T cell populations.(A) Naive, TIM and Flu-TM CD8 T cells were stained with the indicated antibodies (black histogram) or isotype control (filled gray histogram). Data show expression on gated F5 CD8 T cells and are representative of five independent experiments. (B) Polyfunctionality index (calculated as described in the Methods) of specific CD8 T cells was assessed for the simultaneous production of 4 cytokines: IFN-γ, IL-2, CCL3 and CCL5. Data show one out of five independent experiments with similar results. (C) Experimental plan and (D) survival curves of infected mice. Mice were transferred with 5 × 104 naive or TIM or Flu-TM CD8 T cells and infected intranasally 48 hours later with a lethal dose of Flu-NP68 (1 × 107 TCID50). Data are mean of 2 independent experiments. Total number of mice is indicated in brackets. Survival was significantly improved in the Flu-TM group as compared with naive group (*p < 0.05 Log-rank test).
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f1: Comparison of phenotype and protective capacity between two memory CD8 T cell populations.(A) Naive, TIM and Flu-TM CD8 T cells were stained with the indicated antibodies (black histogram) or isotype control (filled gray histogram). Data show expression on gated F5 CD8 T cells and are representative of five independent experiments. (B) Polyfunctionality index (calculated as described in the Methods) of specific CD8 T cells was assessed for the simultaneous production of 4 cytokines: IFN-γ, IL-2, CCL3 and CCL5. Data show one out of five independent experiments with similar results. (C) Experimental plan and (D) survival curves of infected mice. Mice were transferred with 5 × 104 naive or TIM or Flu-TM CD8 T cells and infected intranasally 48 hours later with a lethal dose of Flu-NP68 (1 × 107 TCID50). Data are mean of 2 independent experiments. Total number of mice is indicated in brackets. Survival was significantly improved in the Flu-TM group as compared with naive group (*p < 0.05 Log-rank test).

Mentions: We first compared the phenotype and the protective capacity of two populations of memory cells generated using naive F5 TCR transgenic cells that are specific for NP68 epitope. Antigen-specific TIM cells were generated in a sterile inflammatory context by direct immunization of F5 TCR transgenic mice with the antigenic NP68 peptide in saline202122. Flu-induced memory cells (Flu-TM) were generated by intranasal infection with influenza virus expressing the NP68 epitope of C57BL/6 J mice that were grafted with 2 × 105 naive F5 CD8 T cells. Although this represents a substantial number of transferred naive CD8 T cells, the initial frequency of naive TCR-transgenic CD8 T cells does not influence functional properties of memory T cells and their ability to protect from re-challenge1323. In terms of phenotype and cytokine secretion capacity, TIM memory cells, similarly to Flu-TM F5 cells, display a prototypic memory phenotype expressing increased levels of CD44, CXCR3 and CD122 that distinguish them from naive cells (Fig. 1A) and show significant polyfunctionality (Fig. 1B). We next measured the capacity of the different subsets of CD8 T cells to protect mice against a lethal dose of influenza virus. Naive C57BL/6 J mice were grafted with identical numbers of splenic naive, TIM or Flu-TM F5 CD8 T cells before being intra-nasally infected with a lethal dose of influenza virus (Fig. 1C). Results in Fig. 1D show that the transfer of Flu-TM confers a significant protection against a lethal dose of virus, compared to naive mice. This is in contrast to what is observed with TIM F5 cells. Hence, although TIM and Flu-TM share a memory phenotype, only Flu-TM cells display intrinsic functional capacities, acquired following their priming by the virus and allowing them to curtail a lethal infection by influenza virus.


Immune signatures of protective spleen memory CD8 T cells
Comparison of phenotype and protective capacity between two memory CD8 T cell populations.(A) Naive, TIM and Flu-TM CD8 T cells were stained with the indicated antibodies (black histogram) or isotype control (filled gray histogram). Data show expression on gated F5 CD8 T cells and are representative of five independent experiments. (B) Polyfunctionality index (calculated as described in the Methods) of specific CD8 T cells was assessed for the simultaneous production of 4 cytokines: IFN-γ, IL-2, CCL3 and CCL5. Data show one out of five independent experiments with similar results. (C) Experimental plan and (D) survival curves of infected mice. Mice were transferred with 5 × 104 naive or TIM or Flu-TM CD8 T cells and infected intranasally 48 hours later with a lethal dose of Flu-NP68 (1 × 107 TCID50). Data are mean of 2 independent experiments. Total number of mice is indicated in brackets. Survival was significantly improved in the Flu-TM group as compared with naive group (*p < 0.05 Log-rank test).
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f1: Comparison of phenotype and protective capacity between two memory CD8 T cell populations.(A) Naive, TIM and Flu-TM CD8 T cells were stained with the indicated antibodies (black histogram) or isotype control (filled gray histogram). Data show expression on gated F5 CD8 T cells and are representative of five independent experiments. (B) Polyfunctionality index (calculated as described in the Methods) of specific CD8 T cells was assessed for the simultaneous production of 4 cytokines: IFN-γ, IL-2, CCL3 and CCL5. Data show one out of five independent experiments with similar results. (C) Experimental plan and (D) survival curves of infected mice. Mice were transferred with 5 × 104 naive or TIM or Flu-TM CD8 T cells and infected intranasally 48 hours later with a lethal dose of Flu-NP68 (1 × 107 TCID50). Data are mean of 2 independent experiments. Total number of mice is indicated in brackets. Survival was significantly improved in the Flu-TM group as compared with naive group (*p < 0.05 Log-rank test).
Mentions: We first compared the phenotype and the protective capacity of two populations of memory cells generated using naive F5 TCR transgenic cells that are specific for NP68 epitope. Antigen-specific TIM cells were generated in a sterile inflammatory context by direct immunization of F5 TCR transgenic mice with the antigenic NP68 peptide in saline202122. Flu-induced memory cells (Flu-TM) were generated by intranasal infection with influenza virus expressing the NP68 epitope of C57BL/6 J mice that were grafted with 2 × 105 naive F5 CD8 T cells. Although this represents a substantial number of transferred naive CD8 T cells, the initial frequency of naive TCR-transgenic CD8 T cells does not influence functional properties of memory T cells and their ability to protect from re-challenge1323. In terms of phenotype and cytokine secretion capacity, TIM memory cells, similarly to Flu-TM F5 cells, display a prototypic memory phenotype expressing increased levels of CD44, CXCR3 and CD122 that distinguish them from naive cells (Fig. 1A) and show significant polyfunctionality (Fig. 1B). We next measured the capacity of the different subsets of CD8 T cells to protect mice against a lethal dose of influenza virus. Naive C57BL/6 J mice were grafted with identical numbers of splenic naive, TIM or Flu-TM F5 CD8 T cells before being intra-nasally infected with a lethal dose of influenza virus (Fig. 1C). Results in Fig. 1D show that the transfer of Flu-TM confers a significant protection against a lethal dose of virus, compared to naive mice. This is in contrast to what is observed with TIM F5 cells. Hence, although TIM and Flu-TM share a memory phenotype, only Flu-TM cells display intrinsic functional capacities, acquired following their priming by the virus and allowing them to curtail a lethal infection by influenza virus.

View Article: PubMed Central - PubMed

ABSTRACT

Memory CD8 T lymphocyte populations are remarkably heterogeneous and differ in their ability to protect the host. In order to identify the whole range of qualities uniquely associated with protective memory cells we compared the gene expression signatures of two qualities of memory CD8 T cells sharing the same antigenic-specificity: protective (Influenza-induced, Flu-TM) and non-protective (peptide-induced, TIM) spleen memory CD8 T cells. Although Flu-TM and TIM express classical phenotypic memory markers and are polyfunctional, only Flu-TM protects against a lethal viral challenge. Protective memory CD8 T cells express a unique set of genes involved in migration and survival that correlate with their unique capacity to rapidly migrate within the infected lung parenchyma in response to influenza infection. We also enlighten a new set of poised genes expressed by protective cells that is strongly enriched in cytokines and chemokines such as Ccl1, Ccl9 and Gm-csf. CCL1 and GM-CSF genes are also poised in human memory CD8 T cells. These immune signatures are also induced by two other pathogens (vaccinia virus and Listeria monocytogenes). The immune signatures associated with immune protection were identified on circulating cells, i.e. those that are easily accessible for immuno-monitoring and could help predict vaccines efficacy.

No MeSH data available.