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T cells and T cell tumors efficiently generate antigen-specific cytotoxic T cell immunity when modified with an NKT ligand.

Chung Y, Lee YH, Zhang Y, Martin-Orozco N, Yamazaki T, Zhou D, Kang CY, Hwu P, Kwak LW, Dong C - Oncoimmunology (2012)

Bottom Line: While T cells loaded with a class I-restricted peptide induced proliferation but not effector differentiation of antigen-specific CD8 T cells, injection of T cells co-pulsed with αGC strongly induced IFNγ and Granzyme B expression in T cells and complete lysis of target cells in vivo.Of note, the generation of this cytotoxic T cell response was independent of IL-4, IFNγ, IL-12, IL-21 and costimulation.Our data indicate that iNKT cell can license a non-professional APC to directly trigger antigen-specific cytotoxic T cell responses, which provides an alternative cellular vaccine strategy against tumors.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology; Center for Cancer Immunology Research; University of Texas MD Anderson Cancer Center; Houston, TX USA ; Institute of Molecular Medicine; University of Texas Medical School; Houston, TX USA.

ABSTRACT
Various Invariant NKT (iNKT) cell ligands have been shown as potent adjuvants in boosting T cell reactivates to antigens on professional APC. Non-professional APC, such as T cells, also co-expressing MHC class I and CD1d, have been unattractive cell vaccine carriers due to their poor immunogenicity. Here, we report that T cells as well as T cell lymphoma can efficiently generate antigen-specific cytotoxic T lymphocytes (CTL) responses in mice in vivo, when formulated to present iNKT ligand α-galactosylceramide (αGC) on their surface CD1d. Vaccination with αGC-pulsed EG-7 T-cell lymphoma induced tumor-specific CTL response and suppressed the growth of EG-7 in a CD8 T cell-dependent manner. Injection of αGC-loaded CD4 T cells in mice efficiently activated iNKT cells in vivo. While T cells loaded with a class I-restricted peptide induced proliferation but not effector differentiation of antigen-specific CD8 T cells, injection of T cells co-pulsed with αGC strongly induced IFNγ and Granzyme B expression in T cells and complete lysis of target cells in vivo. Presentation of αGC and peptide on the same cells was required for optimal CTL response and vaccinating T cells appeared to directly stimulate both iNKT and cytotoxic CD8 T cells. Of note, the generation of this cytotoxic T cell response was independent of IL-4, IFNγ, IL-12, IL-21 and costimulation. Our data indicate that iNKT cell can license a non-professional APC to directly trigger antigen-specific cytotoxic T cell responses, which provides an alternative cellular vaccine strategy against tumors.

No MeSH data available.


Related in: MedlinePlus

Figure 1. Vaccination with αGC-loaded EG-7 generated preventive antitumor activity. (A–D) EG-7 cells were co-cultured overnight with 1 μg/ml of αGC (EG-7/αGC) or vehicle followed by irradiation (50 Gy). C57BL/6 mice (n = 7 per group) were vaccinated with EG-7/veh, EG-7/αGC or untreated (Nil). One week later, all mice were subcutaneously injected with 1 × 106 live EG-7 cells and the survival (A) and tumor size (B) were checked. (C) At the 3 weeks later, splenocytes were isolated from tumor bearing mice and analyzed by flow cytometer after staining with anti-CD11b, anti-Gr1, anti-CD19, anti-CD4 and anti-CD8 antibodies. (D) Splenocytes were restimulated with SIINFEKL in the presence of Golgi-Plug for 5 h before intracellular staining of IFNγ on OVA specific T cells. *p < 0.05, **p < 0.005, p values were calculated with 2-way ANOVA (A), Kaplan-Meier method (B) or Student's t-tests (C and D) in comparison with the EG-7/veh group.
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Figure 1: Figure 1. Vaccination with αGC-loaded EG-7 generated preventive antitumor activity. (A–D) EG-7 cells were co-cultured overnight with 1 μg/ml of αGC (EG-7/αGC) or vehicle followed by irradiation (50 Gy). C57BL/6 mice (n = 7 per group) were vaccinated with EG-7/veh, EG-7/αGC or untreated (Nil). One week later, all mice were subcutaneously injected with 1 × 106 live EG-7 cells and the survival (A) and tumor size (B) were checked. (C) At the 3 weeks later, splenocytes were isolated from tumor bearing mice and analyzed by flow cytometer after staining with anti-CD11b, anti-Gr1, anti-CD19, anti-CD4 and anti-CD8 antibodies. (D) Splenocytes were restimulated with SIINFEKL in the presence of Golgi-Plug for 5 h before intracellular staining of IFNγ on OVA specific T cells. *p < 0.05, **p < 0.005, p values were calculated with 2-way ANOVA (A), Kaplan-Meier method (B) or Student's t-tests (C and D) in comparison with the EG-7/veh group.

Mentions: We first asked whether vaccination with tumor cells of T cell origin can trigger anti-tumor activity, when manipulated to present αGC. To track tumor-specific T cell responses more efficiently in vivo, we employed ovalbumin-expressing EL4 thymoma (EG-7) as our tumor model. To address protective antitumor activity by vaccination with αGC-loaded EG-7, we vaccinated mice with irradiated EG-7 co-cultured with 1 μg/mL of αGC overnight. EG-7 cells incubated with the same volume of solvent (5 μl/mL of 0.5% polysorbate) were used as control (EG-7/veh). After extensive washing, these cells were i.v. injected into syngenic naïve mice. These mice were subcutaneously implanted with live EG-7 and the tumor volume and survival were monitored daily. Mice were sacrificed when tumor diameters reached 20 mm. As depicted in Figure 1A and B, the vaccination with EG-7/αGC significantly reduced the growth of solid tumors and induced prolonged survival in comparison to those of EG-7/veh group. Tumors affect myelopoiesis and induce the expansion of CD11b+Gr-1+ myeloid-derived suppressor cell (MDSC) in the bone marrow, blood, spleen.14 Of note, we also observed a great decrease of CD11b+Gr-1+ population in the spleen of EG-7/αGC vaccinated mice (< 4%) compared with that of EG-7/veh group (> 20%) (Fig. 1C), another indicative of antitumor activity induced by the vaccination. Importantly, mice vaccinated with EG-7/αGC showed higher numbers of CD8+ T cells producing IFNγ upon SIINFEKL stimulation (Fig. 1D). These results overall indicate the induction of anti-tumor activity as well as tumor-specific CD8+ T cell responses upon the vaccination with EG-7/αGC in vivo.


T cells and T cell tumors efficiently generate antigen-specific cytotoxic T cell immunity when modified with an NKT ligand.

Chung Y, Lee YH, Zhang Y, Martin-Orozco N, Yamazaki T, Zhou D, Kang CY, Hwu P, Kwak LW, Dong C - Oncoimmunology (2012)

Figure 1. Vaccination with αGC-loaded EG-7 generated preventive antitumor activity. (A–D) EG-7 cells were co-cultured overnight with 1 μg/ml of αGC (EG-7/αGC) or vehicle followed by irradiation (50 Gy). C57BL/6 mice (n = 7 per group) were vaccinated with EG-7/veh, EG-7/αGC or untreated (Nil). One week later, all mice were subcutaneously injected with 1 × 106 live EG-7 cells and the survival (A) and tumor size (B) were checked. (C) At the 3 weeks later, splenocytes were isolated from tumor bearing mice and analyzed by flow cytometer after staining with anti-CD11b, anti-Gr1, anti-CD19, anti-CD4 and anti-CD8 antibodies. (D) Splenocytes were restimulated with SIINFEKL in the presence of Golgi-Plug for 5 h before intracellular staining of IFNγ on OVA specific T cells. *p < 0.05, **p < 0.005, p values were calculated with 2-way ANOVA (A), Kaplan-Meier method (B) or Student's t-tests (C and D) in comparison with the EG-7/veh group.
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Related In: Results  -  Collection

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Figure 1: Figure 1. Vaccination with αGC-loaded EG-7 generated preventive antitumor activity. (A–D) EG-7 cells were co-cultured overnight with 1 μg/ml of αGC (EG-7/αGC) or vehicle followed by irradiation (50 Gy). C57BL/6 mice (n = 7 per group) were vaccinated with EG-7/veh, EG-7/αGC or untreated (Nil). One week later, all mice were subcutaneously injected with 1 × 106 live EG-7 cells and the survival (A) and tumor size (B) were checked. (C) At the 3 weeks later, splenocytes were isolated from tumor bearing mice and analyzed by flow cytometer after staining with anti-CD11b, anti-Gr1, anti-CD19, anti-CD4 and anti-CD8 antibodies. (D) Splenocytes were restimulated with SIINFEKL in the presence of Golgi-Plug for 5 h before intracellular staining of IFNγ on OVA specific T cells. *p < 0.05, **p < 0.005, p values were calculated with 2-way ANOVA (A), Kaplan-Meier method (B) or Student's t-tests (C and D) in comparison with the EG-7/veh group.
Mentions: We first asked whether vaccination with tumor cells of T cell origin can trigger anti-tumor activity, when manipulated to present αGC. To track tumor-specific T cell responses more efficiently in vivo, we employed ovalbumin-expressing EL4 thymoma (EG-7) as our tumor model. To address protective antitumor activity by vaccination with αGC-loaded EG-7, we vaccinated mice with irradiated EG-7 co-cultured with 1 μg/mL of αGC overnight. EG-7 cells incubated with the same volume of solvent (5 μl/mL of 0.5% polysorbate) were used as control (EG-7/veh). After extensive washing, these cells were i.v. injected into syngenic naïve mice. These mice were subcutaneously implanted with live EG-7 and the tumor volume and survival were monitored daily. Mice were sacrificed when tumor diameters reached 20 mm. As depicted in Figure 1A and B, the vaccination with EG-7/αGC significantly reduced the growth of solid tumors and induced prolonged survival in comparison to those of EG-7/veh group. Tumors affect myelopoiesis and induce the expansion of CD11b+Gr-1+ myeloid-derived suppressor cell (MDSC) in the bone marrow, blood, spleen.14 Of note, we also observed a great decrease of CD11b+Gr-1+ population in the spleen of EG-7/αGC vaccinated mice (< 4%) compared with that of EG-7/veh group (> 20%) (Fig. 1C), another indicative of antitumor activity induced by the vaccination. Importantly, mice vaccinated with EG-7/αGC showed higher numbers of CD8+ T cells producing IFNγ upon SIINFEKL stimulation (Fig. 1D). These results overall indicate the induction of anti-tumor activity as well as tumor-specific CD8+ T cell responses upon the vaccination with EG-7/αGC in vivo.

Bottom Line: While T cells loaded with a class I-restricted peptide induced proliferation but not effector differentiation of antigen-specific CD8 T cells, injection of T cells co-pulsed with αGC strongly induced IFNγ and Granzyme B expression in T cells and complete lysis of target cells in vivo.Of note, the generation of this cytotoxic T cell response was independent of IL-4, IFNγ, IL-12, IL-21 and costimulation.Our data indicate that iNKT cell can license a non-professional APC to directly trigger antigen-specific cytotoxic T cell responses, which provides an alternative cellular vaccine strategy against tumors.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology; Center for Cancer Immunology Research; University of Texas MD Anderson Cancer Center; Houston, TX USA ; Institute of Molecular Medicine; University of Texas Medical School; Houston, TX USA.

ABSTRACT
Various Invariant NKT (iNKT) cell ligands have been shown as potent adjuvants in boosting T cell reactivates to antigens on professional APC. Non-professional APC, such as T cells, also co-expressing MHC class I and CD1d, have been unattractive cell vaccine carriers due to their poor immunogenicity. Here, we report that T cells as well as T cell lymphoma can efficiently generate antigen-specific cytotoxic T lymphocytes (CTL) responses in mice in vivo, when formulated to present iNKT ligand α-galactosylceramide (αGC) on their surface CD1d. Vaccination with αGC-pulsed EG-7 T-cell lymphoma induced tumor-specific CTL response and suppressed the growth of EG-7 in a CD8 T cell-dependent manner. Injection of αGC-loaded CD4 T cells in mice efficiently activated iNKT cells in vivo. While T cells loaded with a class I-restricted peptide induced proliferation but not effector differentiation of antigen-specific CD8 T cells, injection of T cells co-pulsed with αGC strongly induced IFNγ and Granzyme B expression in T cells and complete lysis of target cells in vivo. Presentation of αGC and peptide on the same cells was required for optimal CTL response and vaccinating T cells appeared to directly stimulate both iNKT and cytotoxic CD8 T cells. Of note, the generation of this cytotoxic T cell response was independent of IL-4, IFNγ, IL-12, IL-21 and costimulation. Our data indicate that iNKT cell can license a non-professional APC to directly trigger antigen-specific cytotoxic T cell responses, which provides an alternative cellular vaccine strategy against tumors.

No MeSH data available.


Related in: MedlinePlus