INTERLEUKIN-2-PRIMED NAÏVE CD8 T CELLS AS A POTENTIAL REMEDY FOR OVERCOMING TUMOR-DERIVED IMMUNOSUPPRESSION IN TUMOR MICROENVIRONMENT
(Abstract release date: 05/19/16)
EHA Library. Yang D. 06/09/16; 132667; E1118
Prof. Deok-Hwan Yang
Contributions
Contributions
Abstract
Abstract: E1118
Type: Eposter Presentation
Background
CD8 T cells have cytotoxic effector functions upon antigen stimulation and receive considerable attention for adoptive immunotherapy against cancer. Despite of their potential implication, however, conventional approaches using in vitro expanded CD8 T cells have not been successful so far, mostly due to lack of functionality with cellular exhaustion.
Aims
In this study, we therefore investigated the phenotypic and functional differences among in vitro activated CD8 T cells of three different sources, namely naïve, memory and tumor-infiltrating lymphocytes (TILs) of human and mice, for better understanding mechanisms behind potent effector functions and overcoming limitations of the current approaches.
Methods
Effector T cell populations were activated from CD8+ naïve, memory T cells, and CD8+ TILs obtained from cancer tissue (Teff N, Teff M, and re-stimulated TILs, respectively). Individual effectors were analyzed for functional difference and cytotoxicity. In murine model, we also checked the same methods for the phenotypes of T cell exhaustion and tumor challenging test using EG7-EL4 cell line (expressing the OVAp) in adoptively transferred OT-1, thy1.1 Teff N, Teff M and activated TILs T to C57BL6 mice.
Results
In vitro stimulation of human naïve, memory, and TIL CD8+ T cells using typical anti-CD3/CD28 antibodies were able to generate phenotypically homogenous populations of CD62LloCD44hiOX40hiCD27hi cells. Notably, proliferation and total yield after the in vitro expansion were far greater in NTeff cells than for MTeff and TILeff cells, which was well correlated with the enhanced telomere length of NTeff cells compared to the latter two populations. In line with the longer telomere length, NTeff cells exhibited significantly less amounts of T cell exhaustion markers, PD-1, CTLA-4 and KLRG-1, than those of MTeff and TILeff cells, and moreover, acquired distinct expression patterns of memory-promoting transcription factors, with T-bet and Eomes being highly induced in a rapid, sustainable manner. With regard to tumor-induced immune-suppression, interestingly, NTeff cells appeared to have lower expression of Foxp1, a transcription factor promoting T cell quiescence, and were refractory to apoptosis upon TGF-β conditioning, implying a better survival potential. As with in vitro stimulated human T cells, in vitro priming of murine naïve CD8 T cells using high doses of IL2 were also found to be effective for generating potent effector cells that can exert tumor-specific cytotoxic activity.
Conclusion
Of CD8 T cell pools that are different in their activation states, naïve cells other than pre-existing effector/memory and TIL cells were better poised to develop into potent effector cells after in vitro stimulation, which provide an implication for rational design of adoptive immunotherapy against cancer.
Session topic: E-poster
Keyword(s): CD8 T cells, Cytotoxicity, Tumor immunology
Type: Eposter Presentation
Background
CD8 T cells have cytotoxic effector functions upon antigen stimulation and receive considerable attention for adoptive immunotherapy against cancer. Despite of their potential implication, however, conventional approaches using in vitro expanded CD8 T cells have not been successful so far, mostly due to lack of functionality with cellular exhaustion.
Aims
In this study, we therefore investigated the phenotypic and functional differences among in vitro activated CD8 T cells of three different sources, namely naïve, memory and tumor-infiltrating lymphocytes (TILs) of human and mice, for better understanding mechanisms behind potent effector functions and overcoming limitations of the current approaches.
Methods
Effector T cell populations were activated from CD8+ naïve, memory T cells, and CD8+ TILs obtained from cancer tissue (Teff N, Teff M, and re-stimulated TILs, respectively). Individual effectors were analyzed for functional difference and cytotoxicity. In murine model, we also checked the same methods for the phenotypes of T cell exhaustion and tumor challenging test using EG7-EL4 cell line (expressing the OVAp) in adoptively transferred OT-1, thy1.1 Teff N, Teff M and activated TILs T to C57BL6 mice.
Results
In vitro stimulation of human naïve, memory, and TIL CD8+ T cells using typical anti-CD3/CD28 antibodies were able to generate phenotypically homogenous populations of CD62LloCD44hiOX40hiCD27hi cells. Notably, proliferation and total yield after the in vitro expansion were far greater in NTeff cells than for MTeff and TILeff cells, which was well correlated with the enhanced telomere length of NTeff cells compared to the latter two populations. In line with the longer telomere length, NTeff cells exhibited significantly less amounts of T cell exhaustion markers, PD-1, CTLA-4 and KLRG-1, than those of MTeff and TILeff cells, and moreover, acquired distinct expression patterns of memory-promoting transcription factors, with T-bet and Eomes being highly induced in a rapid, sustainable manner. With regard to tumor-induced immune-suppression, interestingly, NTeff cells appeared to have lower expression of Foxp1, a transcription factor promoting T cell quiescence, and were refractory to apoptosis upon TGF-β conditioning, implying a better survival potential. As with in vitro stimulated human T cells, in vitro priming of murine naïve CD8 T cells using high doses of IL2 were also found to be effective for generating potent effector cells that can exert tumor-specific cytotoxic activity.
Conclusion
Of CD8 T cell pools that are different in their activation states, naïve cells other than pre-existing effector/memory and TIL cells were better poised to develop into potent effector cells after in vitro stimulation, which provide an implication for rational design of adoptive immunotherapy against cancer.
Session topic: E-poster
Keyword(s): CD8 T cells, Cytotoxicity, Tumor immunology
Abstract: E1118
Type: Eposter Presentation
Background
CD8 T cells have cytotoxic effector functions upon antigen stimulation and receive considerable attention for adoptive immunotherapy against cancer. Despite of their potential implication, however, conventional approaches using in vitro expanded CD8 T cells have not been successful so far, mostly due to lack of functionality with cellular exhaustion.
Aims
In this study, we therefore investigated the phenotypic and functional differences among in vitro activated CD8 T cells of three different sources, namely naïve, memory and tumor-infiltrating lymphocytes (TILs) of human and mice, for better understanding mechanisms behind potent effector functions and overcoming limitations of the current approaches.
Methods
Effector T cell populations were activated from CD8+ naïve, memory T cells, and CD8+ TILs obtained from cancer tissue (Teff N, Teff M, and re-stimulated TILs, respectively). Individual effectors were analyzed for functional difference and cytotoxicity. In murine model, we also checked the same methods for the phenotypes of T cell exhaustion and tumor challenging test using EG7-EL4 cell line (expressing the OVAp) in adoptively transferred OT-1, thy1.1 Teff N, Teff M and activated TILs T to C57BL6 mice.
Results
In vitro stimulation of human naïve, memory, and TIL CD8+ T cells using typical anti-CD3/CD28 antibodies were able to generate phenotypically homogenous populations of CD62LloCD44hiOX40hiCD27hi cells. Notably, proliferation and total yield after the in vitro expansion were far greater in NTeff cells than for MTeff and TILeff cells, which was well correlated with the enhanced telomere length of NTeff cells compared to the latter two populations. In line with the longer telomere length, NTeff cells exhibited significantly less amounts of T cell exhaustion markers, PD-1, CTLA-4 and KLRG-1, than those of MTeff and TILeff cells, and moreover, acquired distinct expression patterns of memory-promoting transcription factors, with T-bet and Eomes being highly induced in a rapid, sustainable manner. With regard to tumor-induced immune-suppression, interestingly, NTeff cells appeared to have lower expression of Foxp1, a transcription factor promoting T cell quiescence, and were refractory to apoptosis upon TGF-β conditioning, implying a better survival potential. As with in vitro stimulated human T cells, in vitro priming of murine naïve CD8 T cells using high doses of IL2 were also found to be effective for generating potent effector cells that can exert tumor-specific cytotoxic activity.
Conclusion
Of CD8 T cell pools that are different in their activation states, naïve cells other than pre-existing effector/memory and TIL cells were better poised to develop into potent effector cells after in vitro stimulation, which provide an implication for rational design of adoptive immunotherapy against cancer.
Session topic: E-poster
Keyword(s): CD8 T cells, Cytotoxicity, Tumor immunology
Type: Eposter Presentation
Background
CD8 T cells have cytotoxic effector functions upon antigen stimulation and receive considerable attention for adoptive immunotherapy against cancer. Despite of their potential implication, however, conventional approaches using in vitro expanded CD8 T cells have not been successful so far, mostly due to lack of functionality with cellular exhaustion.
Aims
In this study, we therefore investigated the phenotypic and functional differences among in vitro activated CD8 T cells of three different sources, namely naïve, memory and tumor-infiltrating lymphocytes (TILs) of human and mice, for better understanding mechanisms behind potent effector functions and overcoming limitations of the current approaches.
Methods
Effector T cell populations were activated from CD8+ naïve, memory T cells, and CD8+ TILs obtained from cancer tissue (Teff N, Teff M, and re-stimulated TILs, respectively). Individual effectors were analyzed for functional difference and cytotoxicity. In murine model, we also checked the same methods for the phenotypes of T cell exhaustion and tumor challenging test using EG7-EL4 cell line (expressing the OVAp) in adoptively transferred OT-1, thy1.1 Teff N, Teff M and activated TILs T to C57BL6 mice.
Results
In vitro stimulation of human naïve, memory, and TIL CD8+ T cells using typical anti-CD3/CD28 antibodies were able to generate phenotypically homogenous populations of CD62LloCD44hiOX40hiCD27hi cells. Notably, proliferation and total yield after the in vitro expansion were far greater in NTeff cells than for MTeff and TILeff cells, which was well correlated with the enhanced telomere length of NTeff cells compared to the latter two populations. In line with the longer telomere length, NTeff cells exhibited significantly less amounts of T cell exhaustion markers, PD-1, CTLA-4 and KLRG-1, than those of MTeff and TILeff cells, and moreover, acquired distinct expression patterns of memory-promoting transcription factors, with T-bet and Eomes being highly induced in a rapid, sustainable manner. With regard to tumor-induced immune-suppression, interestingly, NTeff cells appeared to have lower expression of Foxp1, a transcription factor promoting T cell quiescence, and were refractory to apoptosis upon TGF-β conditioning, implying a better survival potential. As with in vitro stimulated human T cells, in vitro priming of murine naïve CD8 T cells using high doses of IL2 were also found to be effective for generating potent effector cells that can exert tumor-specific cytotoxic activity.
Conclusion
Of CD8 T cell pools that are different in their activation states, naïve cells other than pre-existing effector/memory and TIL cells were better poised to develop into potent effector cells after in vitro stimulation, which provide an implication for rational design of adoptive immunotherapy against cancer.
Session topic: E-poster
Keyword(s): CD8 T cells, Cytotoxicity, Tumor immunology
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