Limits...
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

Alteration in numbers and phenotype of DCs in TDLNs after cryo-ablation in glioma mice models.Notes: (A) Scheme to study the alteration of DCs in glioma mice models. GL261 cells (106) were implanted intradermally in the left flank 14 days prior to day 0. Two weeks later, when tumors grew to 5 to 9 mm, mice were randomized into tumor bearing group, cryo-ablation group, and surgery group. After 14 days, TDLNs were removed and DCs were isolated. The second tumors were implanted on day 28. (B, C) Expression levels of CD80, CD86, and MHC class II in gated CD11c+ DCs from four groups were evaluated by FACS analysis. Grey curves represent staining with the isotype-matched control monoclonal antibodies. (D) Absolute amounts of DCs were calculated. 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. #Day –14 is 14 days prior to day 0.Abbreviations: DCs, dendritic cells; TDLNs, tumor-draining lymph nodes; DLN, draining lymph node; FACS, fluorescence activated cell sorting; ANOVA, analysis of variance; SEM, standard error of the mean; MHC, major histocompatibility complex; FITC, fluorescein isothiocyanate.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4362656&req=5

f1-dddt-9-1449: Alteration in numbers and phenotype of DCs in TDLNs after cryo-ablation in glioma mice models.Notes: (A) Scheme to study the alteration of DCs in glioma mice models. GL261 cells (106) were implanted intradermally in the left flank 14 days prior to day 0. Two weeks later, when tumors grew to 5 to 9 mm, mice were randomized into tumor bearing group, cryo-ablation group, and surgery group. After 14 days, TDLNs were removed and DCs were isolated. The second tumors were implanted on day 28. (B, C) Expression levels of CD80, CD86, and MHC class II in gated CD11c+ DCs from four groups were evaluated by FACS analysis. Grey curves represent staining with the isotype-matched control monoclonal antibodies. (D) Absolute amounts of DCs were calculated. 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. #Day –14 is 14 days prior to day 0.Abbreviations: DCs, dendritic cells; TDLNs, tumor-draining lymph nodes; DLN, draining lymph node; FACS, fluorescence activated cell sorting; ANOVA, analysis of variance; SEM, standard error of the mean; MHC, major histocompatibility complex; FITC, fluorescein isothiocyanate.

Mentions: Tumors have been observed to inhibit DC maturation which is considered the initiator of tumor immunity.18 Therefore, this study aims to test whether cryo-ablation could induce tumor immunity through influence on DCs in glioma mice models. At first, we injected GL261 glioma cells into the left flank of C57BL/6 mice 14 days prior to day 0 and performed cryo-ablation or surgery when the tumor diameters reached 5 to 9 mm. Fourteen days after treatment, mice were killed to remove the TDLNs according to mice anatomy atlas (Figure 1A). We isolated the DCs in TDLNs from tumor bearing group, tumor free group, cryo-ablation group, and surgery group to quantify the expression of MHC class II as well as CD86 and CD80 (Figure 1B). Compared with the medium levels of CD86 and CD80 expressed in the tumor free group, DCs isolated from the tumor bearing group expressed low levels of CD86 and CD80. Specifically, DCs from mice whom had their tumors removed 2 weeks prior did not substantially recover the expression of CD86 and CD80, remaining at a medium level. But DCs from the cryo-ablation group had high levels of restored CD86 and CD80. DCs from all groups expressed similar MHC class II molecules (Figure 1C). We also counted the absolute numbers of DCs from TDLNs. There is no significant difference in the numbers of DCs between the tumor bearing group and the surgery group. But DCs in the cryo-ablation group significantly outnumbered the other groups (Figure 1D).


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)

Alteration in numbers and phenotype of DCs in TDLNs after cryo-ablation in glioma mice models.Notes: (A) Scheme to study the alteration of DCs in glioma mice models. GL261 cells (106) were implanted intradermally in the left flank 14 days prior to day 0. Two weeks later, when tumors grew to 5 to 9 mm, mice were randomized into tumor bearing group, cryo-ablation group, and surgery group. After 14 days, TDLNs were removed and DCs were isolated. The second tumors were implanted on day 28. (B, C) Expression levels of CD80, CD86, and MHC class II in gated CD11c+ DCs from four groups were evaluated by FACS analysis. Grey curves represent staining with the isotype-matched control monoclonal antibodies. (D) Absolute amounts of DCs were calculated. 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. #Day –14 is 14 days prior to day 0.Abbreviations: DCs, dendritic cells; TDLNs, tumor-draining lymph nodes; DLN, draining lymph node; FACS, fluorescence activated cell sorting; ANOVA, analysis of variance; SEM, standard error of the mean; MHC, major histocompatibility complex; FITC, fluorescein isothiocyanate.
© Copyright Policy
Related In: Results  -  Collection

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

f1-dddt-9-1449: Alteration in numbers and phenotype of DCs in TDLNs after cryo-ablation in glioma mice models.Notes: (A) Scheme to study the alteration of DCs in glioma mice models. GL261 cells (106) were implanted intradermally in the left flank 14 days prior to day 0. Two weeks later, when tumors grew to 5 to 9 mm, mice were randomized into tumor bearing group, cryo-ablation group, and surgery group. After 14 days, TDLNs were removed and DCs were isolated. The second tumors were implanted on day 28. (B, C) Expression levels of CD80, CD86, and MHC class II in gated CD11c+ DCs from four groups were evaluated by FACS analysis. Grey curves represent staining with the isotype-matched control monoclonal antibodies. (D) Absolute amounts of DCs were calculated. 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. #Day –14 is 14 days prior to day 0.Abbreviations: DCs, dendritic cells; TDLNs, tumor-draining lymph nodes; DLN, draining lymph node; FACS, fluorescence activated cell sorting; ANOVA, analysis of variance; SEM, standard error of the mean; MHC, major histocompatibility complex; FITC, fluorescein isothiocyanate.
Mentions: Tumors have been observed to inhibit DC maturation which is considered the initiator of tumor immunity.18 Therefore, this study aims to test whether cryo-ablation could induce tumor immunity through influence on DCs in glioma mice models. At first, we injected GL261 glioma cells into the left flank of C57BL/6 mice 14 days prior to day 0 and performed cryo-ablation or surgery when the tumor diameters reached 5 to 9 mm. Fourteen days after treatment, mice were killed to remove the TDLNs according to mice anatomy atlas (Figure 1A). We isolated the DCs in TDLNs from tumor bearing group, tumor free group, cryo-ablation group, and surgery group to quantify the expression of MHC class II as well as CD86 and CD80 (Figure 1B). Compared with the medium levels of CD86 and CD80 expressed in the tumor free group, DCs isolated from the tumor bearing group expressed low levels of CD86 and CD80. Specifically, DCs from mice whom had their tumors removed 2 weeks prior did not substantially recover the expression of CD86 and CD80, remaining at a medium level. But DCs from the cryo-ablation group had high levels of restored CD86 and CD80. DCs from all groups expressed similar MHC class II molecules (Figure 1C). We also counted the absolute numbers of DCs from TDLNs. There is no significant difference in the numbers of DCs between the tumor bearing group and the surgery group. But DCs in the cryo-ablation group significantly outnumbered the other groups (Figure 1D).

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