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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

Cryo-ablation converted the expression of IL-10 in TDLN-DCs.Notes: (A) Expressions of intracellular IL-10 in gated CD11c+ DCs from four groups were evaluated by FACS analysis. The upper left quadrant represents the IL-10+ DCs, upper right quadrant represents the IL-10+CD11c+ DCs, lower right quadrant represents the CD11c+ DCs, the lower left quadrant represents the IL-10− CD11c− DCs. (B) Percentages of IL-10+CD11c+ were calculated in the histogram. *P<0.05 represents the IL-10 levels in the cryo-ablation group compared to tumor bearing or surgery group in (C). (C) Ex vivo IL-10 secretion by 104 DCs in overnight culture, determined by ELISA, presented as the mean plus SEM of triplicate wells. The data were analyzed by Student’s t-test. *P<0.05 represents the IL-10+ DC cells in the cryo-ablation group compared to tumor bearing or surgery group in Figure 2B.Abbreviations: IL, interleukin; TDLN, tumor-draining lymph node; DCs, dendritic cells; FACS, fluorescence activated cell sorting; ELISA, enzyme-linked immunosorbent assay; SEM, standard error of the mean; MHC, major histocompatibility complex; FSC, forward scatter; SSC, side scatter.
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f2-dddt-9-1449: Cryo-ablation converted the expression of IL-10 in TDLN-DCs.Notes: (A) Expressions of intracellular IL-10 in gated CD11c+ DCs from four groups were evaluated by FACS analysis. The upper left quadrant represents the IL-10+ DCs, upper right quadrant represents the IL-10+CD11c+ DCs, lower right quadrant represents the CD11c+ DCs, the lower left quadrant represents the IL-10− CD11c− DCs. (B) Percentages of IL-10+CD11c+ were calculated in the histogram. *P<0.05 represents the IL-10 levels in the cryo-ablation group compared to tumor bearing or surgery group in (C). (C) Ex vivo IL-10 secretion by 104 DCs in overnight culture, determined by ELISA, presented as the mean plus SEM of triplicate wells. The data were analyzed by Student’s t-test. *P<0.05 represents the IL-10+ DC cells in the cryo-ablation group compared to tumor bearing or surgery group in Figure 2B.Abbreviations: IL, interleukin; TDLN, tumor-draining lymph node; DCs, dendritic cells; FACS, fluorescence activated cell sorting; ELISA, enzyme-linked immunosorbent assay; SEM, standard error of the mean; MHC, major histocompatibility complex; FSC, forward scatter; SSC, side scatter.

Mentions: IL-10 was an important immunosuppressive molecule which was rarely expressed in mature DCs but was more often seen in some special DCs such as tumor induced DCs.6 Thus, we examined the IL-10 expression in DCs to evaluate the influence of cryo-ablation. DCs were identified on the gate of CD11c+ from the total TDLNs and then stained with MHC II and intracellular IL-10 (Figure 2A). Almost half of DCs from the tumor bearing group expressed intracellular IL-10, while few DCs from the tumor free group expressed intracellular IL-10. Significantly, DCs from the cryo-ablation group expressed lower intracellular IL-10 than DCs from the surgery group (Figure 2B). There was no relevance between intracellular IL-10 and MHC II. We also tested the secretion of extra-cellular IL-10 in DCs. In line with the expression of intracellular IL-10, DCs from the tumor bearing group secreted more extracellular IL-10 than DCs from the tumor free group, and DCs from the cryo-ablation group secreted less IL-10 than DCs from the surgery group (Figure 2C).


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)

Cryo-ablation converted the expression of IL-10 in TDLN-DCs.Notes: (A) Expressions of intracellular IL-10 in gated CD11c+ DCs from four groups were evaluated by FACS analysis. The upper left quadrant represents the IL-10+ DCs, upper right quadrant represents the IL-10+CD11c+ DCs, lower right quadrant represents the CD11c+ DCs, the lower left quadrant represents the IL-10− CD11c− DCs. (B) Percentages of IL-10+CD11c+ were calculated in the histogram. *P<0.05 represents the IL-10 levels in the cryo-ablation group compared to tumor bearing or surgery group in (C). (C) Ex vivo IL-10 secretion by 104 DCs in overnight culture, determined by ELISA, presented as the mean plus SEM of triplicate wells. The data were analyzed by Student’s t-test. *P<0.05 represents the IL-10+ DC cells in the cryo-ablation group compared to tumor bearing or surgery group in Figure 2B.Abbreviations: IL, interleukin; TDLN, tumor-draining lymph node; DCs, dendritic cells; FACS, fluorescence activated cell sorting; ELISA, enzyme-linked immunosorbent assay; SEM, standard error of the mean; MHC, major histocompatibility complex; FSC, forward scatter; SSC, side scatter.
© Copyright Policy
Related In: Results  -  Collection

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

f2-dddt-9-1449: Cryo-ablation converted the expression of IL-10 in TDLN-DCs.Notes: (A) Expressions of intracellular IL-10 in gated CD11c+ DCs from four groups were evaluated by FACS analysis. The upper left quadrant represents the IL-10+ DCs, upper right quadrant represents the IL-10+CD11c+ DCs, lower right quadrant represents the CD11c+ DCs, the lower left quadrant represents the IL-10− CD11c− DCs. (B) Percentages of IL-10+CD11c+ were calculated in the histogram. *P<0.05 represents the IL-10 levels in the cryo-ablation group compared to tumor bearing or surgery group in (C). (C) Ex vivo IL-10 secretion by 104 DCs in overnight culture, determined by ELISA, presented as the mean plus SEM of triplicate wells. The data were analyzed by Student’s t-test. *P<0.05 represents the IL-10+ DC cells in the cryo-ablation group compared to tumor bearing or surgery group in Figure 2B.Abbreviations: IL, interleukin; TDLN, tumor-draining lymph node; DCs, dendritic cells; FACS, fluorescence activated cell sorting; ELISA, enzyme-linked immunosorbent assay; SEM, standard error of the mean; MHC, major histocompatibility complex; FSC, forward scatter; SSC, side scatter.
Mentions: IL-10 was an important immunosuppressive molecule which was rarely expressed in mature DCs but was more often seen in some special DCs such as tumor induced DCs.6 Thus, we examined the IL-10 expression in DCs to evaluate the influence of cryo-ablation. DCs were identified on the gate of CD11c+ from the total TDLNs and then stained with MHC II and intracellular IL-10 (Figure 2A). Almost half of DCs from the tumor bearing group expressed intracellular IL-10, while few DCs from the tumor free group expressed intracellular IL-10. Significantly, DCs from the cryo-ablation group expressed lower intracellular IL-10 than DCs from the surgery group (Figure 2B). There was no relevance between intracellular IL-10 and MHC II. We also tested the secretion of extra-cellular IL-10 in DCs. In line with the expression of intracellular IL-10, DCs from the tumor bearing group secreted more extracellular IL-10 than DCs from the tumor free group, and DCs from the cryo-ablation group secreted less IL-10 than DCs from the surgery group (Figure 2C).

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