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Cryo-ablation improves anti-tumor immunity through recovering tumor educated dendritic cells in tumor-draining lymph nodes.

He XZ, Wang QF, Han S, Wang HQ, Ye YY, Zhu ZY, Zhang SZ - Drug Des Devel Ther (2015)

Bottom Line: Moreover, the DCs decreased the expression of intracellular interleukin-10 (IL-10) and extra-cellular IL-10.More importantly, Tregs inhibited the performance of these DCs; and depletion of Tregs greatly improved anti-tumor immunity in vivo.The Tregs/CD4(+)T and Tregs/CD25(+)T cells in TDLNs inhibit DCs' activity and function.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou, People's Republic of China ; The National Key Clinic Specialty, The Neurosurgery Institute of Guangdong Province, Guangdong Provincial Key Laboratory on Brain Function Repair and Regeneration, Southern Medical University, Guangzhou, People's Republic of China.

ABSTRACT

Background: In addition to minimally invasive destruction of tumors, cryo-ablation of tumors to some extent modulated anti-tumor immunity. Cryo-ablated tumors in glioma mice models induced anti-tumor cellular immunologic response which increases the percentage of CD3(+) and CD4(+)T cells in blood as well as natural killer cells. As a crucial role in triggering anti-tumor immunity, dendritic cells (DCs) were educated by tumors to adopt a tolerance phenotype which helps the tumor escape from immune monitoring. This study aims to study whether cryo-ablation could influence the tolerogenic DCs, and influence anti-tumor immunity in tumor-draining lymph nodes (TDLNs).

Methods: Using the GL261 subcutaneous glioma mouse model, we created a tumor bearing group, cryo-ablation group, and surgery group. We analyzed alteration in phenotype and function of tolerogenic DCs, and evaluated the factors of anti-tumor immunity inhibition.

Results: DCs in TDLNs in GL261 subcutaneous glioma mouse model expressed tolerogenic phenotype. In contrast to surgery, cryo-ablation improved the quantity and quality of these tolerogenic DCs. Moreover, the DCs decreased the expression of intracellular interleukin-10 (IL-10) and extra-cellular IL-10. In vitro, DCs from the cryo-ablation group recovered their specific function and induced potent anti-tumor immunity through triggering T cells. In vivo, cryo-ablation showed weak anti-tumor immunity, only inhibiting the growth of rechallenged tumors. But many IL-10-low DCs, rather than IL-10-high DCs, infiltrated the tumors. More importantly, Tregs inhibited the performance of these DCs; and depletion of Tregs greatly improved anti-tumor immunity in vivo.

Conclusion: Cryo-ablation could recover function of tumor induced tolerogenic DCs in vitro; and depletion of Tregs could improve this anti-tumor effect in vivo. The Tregs/CD4(+)T and Tregs/CD25(+)T cells in TDLNs inhibit DCs' activity and function.

No MeSH data available.


Related in: MedlinePlus

The capacity of DCs to stimulate T cells enhanced after cryo-ablation.Notes: (A) Mixed lymphocyte responses were performed. Responder CD3+T cells were from BALB/c mice, labeled with CFSE, and incubated with the different DCs. Following incubation, the proliferation levels of cells were analyzed by FACS. (B) T cells were proliferated at various DCs:T cells ratios. (C) T cells stimulated with different DCs from four groups showed cytotoxicity to GL261 cells at various E:T ratios. (D) T cells stimulated with these DCs plus GL261 lysates showed cytotoxicity to GL261 cells at various E:T ratios. The data are representative of three independent experiments. Statistical significance was calculated by one-way ANOVA using the Bonferroni post-test with the following notations for P-value. *P<0.05, error bars, SEM.Abbreviations: DCs, dendritic cells; CFSE, carboxyfluorescein diacetate succinimidyl ester; FACS, fluorescence activated cell sorting; ANOVA, analysis of variance; SEM, standard error of the mean; FITC, fluorescein isothiocyanate.
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f3-dddt-9-1449: The capacity of DCs to stimulate T cells enhanced after cryo-ablation.Notes: (A) Mixed lymphocyte responses were performed. Responder CD3+T cells were from BALB/c mice, labeled with CFSE, and incubated with the different DCs. Following incubation, the proliferation levels of cells were analyzed by FACS. (B) T cells were proliferated at various DCs:T cells ratios. (C) T cells stimulated with different DCs from four groups showed cytotoxicity to GL261 cells at various E:T ratios. (D) T cells stimulated with these DCs plus GL261 lysates showed cytotoxicity to GL261 cells at various E:T ratios. The data are representative of three independent experiments. Statistical significance was calculated by one-way ANOVA using the Bonferroni post-test with the following notations for P-value. *P<0.05, error bars, SEM.Abbreviations: DCs, dendritic cells; CFSE, carboxyfluorescein diacetate succinimidyl ester; FACS, fluorescence activated cell sorting; ANOVA, analysis of variance; SEM, standard error of the mean; FITC, fluorescein isothiocyanate.

Mentions: Some reports described that tumor induced DCs had immunosuppressive functions on T cells,6,9 so we performed functional tests to examine whether cryo-ablation could promote immune capacity of DCs in vitro. First, we tested the T cell priming capacity of DCs in an allogeneic proliferation assay. T cells isolated from BALB/c mice spleen were stained with CFSE and showed strong proliferation with anti-CD3 stimulation. Even at high DC:T cell ratio, DCs from the tumor bearing group showed weak capacity of T cell proliferation. DCs from the cryo-ablation group showed stronger T cell proliferation than DCs from the surgery group. The proliferation capacity was proportional with DCs:T cell ratio (Figure 3A and B). Next, we examined whether T cells could induce powerful cytotoxicity to GL261 cells through CTL assay. Only T cells stimulated with DCs from the cryo-ablation group induced significant cytotoxicity to GL261, while T cells stimulated with DCs from the surgery group could not induce powerful cytotoxicity to Gl261 cells even at high E:T ratio (Figure 3C). When we loaded these DCs with GL261 lysates, T cells which were stimulated with DCs from the tumor free group showed higher cytotoxicity than those stimulated with DCs from the cryo-ablation group. But contrary to unloading DCs, T cells which were stimulated with DCs loaded with GL261 lysates did not enhance cytotoxicity of GL261 cells (Figure 3D). Actually, for every cycle of division, a new peak with less intensity of fluorescence appeared, however, we only illustrated this in a histogram (the other cycle of division was not illustrated here). Actually Figure 1A could reflect the capacity of DCs to stimulate T cells enhanced after cryo-ablation. Additionally, Figure 3B and C would be helpful to observe the capacity of DCs stimulating the cytotoxicity of T cells.


Cryo-ablation improves anti-tumor immunity through recovering tumor educated dendritic cells in tumor-draining lymph nodes.

He XZ, Wang QF, Han S, Wang HQ, Ye YY, Zhu ZY, Zhang SZ - Drug Des Devel Ther (2015)

The capacity of DCs to stimulate T cells enhanced after cryo-ablation.Notes: (A) Mixed lymphocyte responses were performed. Responder CD3+T cells were from BALB/c mice, labeled with CFSE, and incubated with the different DCs. Following incubation, the proliferation levels of cells were analyzed by FACS. (B) T cells were proliferated at various DCs:T cells ratios. (C) T cells stimulated with different DCs from four groups showed cytotoxicity to GL261 cells at various E:T ratios. (D) T cells stimulated with these DCs plus GL261 lysates showed cytotoxicity to GL261 cells at various E:T ratios. The data are representative of three independent experiments. Statistical significance was calculated by one-way ANOVA using the Bonferroni post-test with the following notations for P-value. *P<0.05, error bars, SEM.Abbreviations: DCs, dendritic cells; CFSE, carboxyfluorescein diacetate succinimidyl ester; FACS, fluorescence activated cell sorting; ANOVA, analysis of variance; SEM, standard error of the mean; FITC, fluorescein isothiocyanate.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362656&req=5

f3-dddt-9-1449: The capacity of DCs to stimulate T cells enhanced after cryo-ablation.Notes: (A) Mixed lymphocyte responses were performed. Responder CD3+T cells were from BALB/c mice, labeled with CFSE, and incubated with the different DCs. Following incubation, the proliferation levels of cells were analyzed by FACS. (B) T cells were proliferated at various DCs:T cells ratios. (C) T cells stimulated with different DCs from four groups showed cytotoxicity to GL261 cells at various E:T ratios. (D) T cells stimulated with these DCs plus GL261 lysates showed cytotoxicity to GL261 cells at various E:T ratios. The data are representative of three independent experiments. Statistical significance was calculated by one-way ANOVA using the Bonferroni post-test with the following notations for P-value. *P<0.05, error bars, SEM.Abbreviations: DCs, dendritic cells; CFSE, carboxyfluorescein diacetate succinimidyl ester; FACS, fluorescence activated cell sorting; ANOVA, analysis of variance; SEM, standard error of the mean; FITC, fluorescein isothiocyanate.
Mentions: Some reports described that tumor induced DCs had immunosuppressive functions on T cells,6,9 so we performed functional tests to examine whether cryo-ablation could promote immune capacity of DCs in vitro. First, we tested the T cell priming capacity of DCs in an allogeneic proliferation assay. T cells isolated from BALB/c mice spleen were stained with CFSE and showed strong proliferation with anti-CD3 stimulation. Even at high DC:T cell ratio, DCs from the tumor bearing group showed weak capacity of T cell proliferation. DCs from the cryo-ablation group showed stronger T cell proliferation than DCs from the surgery group. The proliferation capacity was proportional with DCs:T cell ratio (Figure 3A and B). Next, we examined whether T cells could induce powerful cytotoxicity to GL261 cells through CTL assay. Only T cells stimulated with DCs from the cryo-ablation group induced significant cytotoxicity to GL261, while T cells stimulated with DCs from the surgery group could not induce powerful cytotoxicity to Gl261 cells even at high E:T ratio (Figure 3C). When we loaded these DCs with GL261 lysates, T cells which were stimulated with DCs from the tumor free group showed higher cytotoxicity than those stimulated with DCs from the cryo-ablation group. But contrary to unloading DCs, T cells which were stimulated with DCs loaded with GL261 lysates did not enhance cytotoxicity of GL261 cells (Figure 3D). Actually, for every cycle of division, a new peak with less intensity of fluorescence appeared, however, we only illustrated this in a histogram (the other cycle of division was not illustrated here). Actually Figure 1A could reflect the capacity of DCs to stimulate T cells enhanced after cryo-ablation. Additionally, Figure 3B and C would be helpful to observe the capacity of DCs stimulating the cytotoxicity of T cells.

Bottom Line: Moreover, the DCs decreased the expression of intracellular interleukin-10 (IL-10) and extra-cellular IL-10.More importantly, Tregs inhibited the performance of these DCs; and depletion of Tregs greatly improved anti-tumor immunity in vivo.The Tregs/CD4(+)T and Tregs/CD25(+)T cells in TDLNs inhibit DCs' activity and function.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou, People's Republic of China ; The National Key Clinic Specialty, The Neurosurgery Institute of Guangdong Province, Guangdong Provincial Key Laboratory on Brain Function Repair and Regeneration, Southern Medical University, Guangzhou, People's Republic of China.

ABSTRACT

Background: In addition to minimally invasive destruction of tumors, cryo-ablation of tumors to some extent modulated anti-tumor immunity. Cryo-ablated tumors in glioma mice models induced anti-tumor cellular immunologic response which increases the percentage of CD3(+) and CD4(+)T cells in blood as well as natural killer cells. As a crucial role in triggering anti-tumor immunity, dendritic cells (DCs) were educated by tumors to adopt a tolerance phenotype which helps the tumor escape from immune monitoring. This study aims to study whether cryo-ablation could influence the tolerogenic DCs, and influence anti-tumor immunity in tumor-draining lymph nodes (TDLNs).

Methods: Using the GL261 subcutaneous glioma mouse model, we created a tumor bearing group, cryo-ablation group, and surgery group. We analyzed alteration in phenotype and function of tolerogenic DCs, and evaluated the factors of anti-tumor immunity inhibition.

Results: DCs in TDLNs in GL261 subcutaneous glioma mouse model expressed tolerogenic phenotype. In contrast to surgery, cryo-ablation improved the quantity and quality of these tolerogenic DCs. Moreover, the DCs decreased the expression of intracellular interleukin-10 (IL-10) and extra-cellular IL-10. In vitro, DCs from the cryo-ablation group recovered their specific function and induced potent anti-tumor immunity through triggering T cells. In vivo, cryo-ablation showed weak anti-tumor immunity, only inhibiting the growth of rechallenged tumors. But many IL-10-low DCs, rather than IL-10-high DCs, infiltrated the tumors. More importantly, Tregs inhibited the performance of these DCs; and depletion of Tregs greatly improved anti-tumor immunity in vivo.

Conclusion: Cryo-ablation could recover function of tumor induced tolerogenic DCs in vitro; and depletion of Tregs could improve this anti-tumor effect in vivo. The Tregs/CD4(+)T and Tregs/CD25(+)T cells in TDLNs inhibit DCs' activity and function.

No MeSH data available.


Related in: MedlinePlus