Dataset Information

Tumor-released autophagosomes induces CD4⁺ T cell-mediated immunosuppression via a TLR2-IL-6 cascade.

ABSTRACT:

Background

CD4⁺ T cells are critical effectors of anti-tumor immunity, but how tumor cells influence CD4⁺ T cell effector function is not fully understood. Tumor cell-released autophagosomes (TRAPs) are being recognized as critical modulators of host anti-tumor immunity during tumor progression. Here, we explored the mechanistic aspects of TRAPs in the modulation of CD4⁺ T cells in the tumor microenvironment.

Methods

TRAPs isolated from tumor cell lines and pleural effusions or ascites of cancer patients were incubated with CD4⁺ T cells to examine the function and mechanism of TRAPs in CD4⁺ T cell differentiation and function. TRAPs-elicited CD4⁺ T cells were tested for their suppression of effector T cell function, induction of regulatory B cells, and promotion of tumorigenesis and metastasis in a mouse model.

Results

Heat shock protein 90α (HSP90α) on the surface of TRAPs from malignant effusions of cancer patients and tumor cell lines stimulated CD4⁺ T cell production of IL-6 via a TLR2-MyD88-NF-κB signal cascade. TRAPs-induced autocrine IL-6 further promoted CD4⁺ T cells secretion of IL-10 and IL-21 via STAT3. Notably, TRAPs-elicited CD4⁺ T cells inhibited CD4⁺ and CD8⁺ effector T cell function in an IL-6- and IL-10-dependent manner and induced IL-10-producing regulatory B cells (Bregs) via IL-6, IL-10 and IL-21, thereby promoting tumor growth and metastasis. Consistently, inhibition of tumor autophagosome formation or IL-6 secretion by CD4⁺ T cells markedly retarded tumor growth. Furthermore, B cell or CD4⁺ T cell depletion impeded tumor growth by increasing effector T cell function.

Conclusions

HSP90α on the surface of TRAPs programs the immunosuppressive functions of CD4⁺ T cells to promote tumor growth and metastasis. TRAPs or their membrane-bound HSP90α represent important therapeutic targets to reverse cancer-associated immunosuppression and improve immunotherapy.

SUBMITTER: Chen YQ

PROVIDER: S-EPMC6625067 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature

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Publications

Tumor-released autophagosomes induces CD4<sup>+</sup> T cell-mediated immunosuppression via a TLR2-IL-6 cascade.

Chen Yong-Qiang YQ Li Peng-Cheng PC Pan Ning N Gao Rong R Wen Zhi-Fa ZF Zhang Tian-Yu TY Huang Fang F Wu Fang-Yuan FY Ou Xi-Long XL Zhang Jin-Ping JP Zhu Xue-Jun XJ Hu Hong-Ming HM Chen Kang K Cai Yun-Lang YL Wang Li-Xin LX

Journal for immunotherapy of cancer 20190712 1

<h4>Background</h4>CD4<sup>+</sup> T cells are critical effectors of anti-tumor immunity, but how tumor cells influence CD4<sup>+</sup> T cell effector function is not fully understood. Tumor cell-released autophagosomes (TRAPs) are being recognized as critical modulators of host anti-tumor immunity during tumor progression. Here, we explored the mechanistic aspects of TRAPs in the modulation of CD4<sup>+</sup> T cells in the tumor microenvironment.<h4>Methods</h4>TRAPs isolated from tumor cell ...[more]

PMID: 31300052

Similar Datasets

Project description:BACKGROUND:Tumor-associated macrophages (TAMs) facilitate tumor progression via establishment of an immunosuppressive tumor microenvironment (TME). However, it is poorly understood how tumor cells could functionally modulate TAMs. Our previous work indicated that tumor cell-released autophagosomes (TRAPs), a type of LC3-II+ double-membrane extracellular vesicles (EVs) was sufficient to suppress anti-tumor immune responses by inducing IL-10-producing B cells and immune suppressive neutrophils. Here, we hypothesized that TRAPs may participate in regulating macrophage polarization. METHODS:TRAPs isolated from multiple murine tumor cell lines and pleural effusions or ascites of cancer patients were incubated with bone marrow-derived macrophages (BMDMs) and monocytes, respectively. Cellular phenotypes were examined by flow cytometry, ELISA and quantitative PCR. TRAPs treated BMDMs were tested for the ability to suppress T-cell proliferation in vitro, and for promotion of tumor growth in vivo. Transwell chamber and neutralization antibodies were added to ascertain the inhibitory molecules expressed on BMDMs exposed to TRAPs. Knockout mice were used to identify the receptors responsible for TRAPs-induced BMDMs polarization and the signaling mechanism was examined by western blot. Autophagy-deficient tumors were profiled for phenotypic changes of TAMs and IFN-γ secretion of T cells by flow cytometry. The phenotype of monocytes from pleural effusions or ascites of cancer patients was assessed by flow cytometry. RESULTS:TRAPs converted macrophages into an immunosuppressive M2-like phenotype characterized by the expression of PD-L1 and IL-10. These macrophages inhibited the proliferation of both CD4+ and CD8+ T cells in vitro, and promoted tumor growth mainly through PD-L1 in vivo. TRAPs-induced macrophage polarization was dependent on TLR4-mediated MyD88-p38-STAT3 signaling. In vivo studies indicated that disruption of autophagosome formation in B16F10 cells by silencing the autophagy gene Beclin1 resulted in a remarkable delay in tumor growth, which was associated with reduced autophagosome secretion, TAMs reprogramming and enhanced T cell activation. Moreover, the levels of LC3B+ EVs appeared to correlate significantly with up-regulation of PD-L1 and IL-10 in matched monocytes from effusions or ascites of cancer patients, and TRAPs isolated from these samples could also polarize monocytes to an M2-like phenotype with increased expression of PD-L1, CD163 and IL-10, decreased expression of HLA-DR, and T cell-suppressive function. CONCLUSIONS:These findings suggest the TRAPs-PD-L1 axis as a major driver of immunosuppression in the TME by eliciting macrophage polarization towards an M2-like phenotype, and highlight the potential novel therapeutic approach of simultaneously targeting autophagy and PD-L1.

Project description:Background: A hallmark of cancer is the presence of an immunosuppressive tumor microenvironment (TME). Immunosuppressive M2 macrophages (MΦs) in the TME facilitate escape from immune surveillance and promote tumor growth; therefore, TME-induced immunosuppression is a potent immunotherapeutic approach to treating cancer. Methods: Cancer cell-secreted proteins were detected by using liquid chromatography-mass spectrometry (LC-MS). Neutralizing antibodies (nAbs) were used to assess which proteins were involved in MΦs polarization and differentiation. The protein-protein interaction was characterized using co-immunoprecipitation and immunofluorescence assays. Cancer-secreted heat shock protein 70 (Hsp70) protein was quantified using an enzyme-linked immunosorbent assay (ELISA). MΦ polarization and tumor growth were assessed in vivo with subcutaneous LLC-GFP tumor models and toll-like receptor 2 (TLR2) knockout mice; in vitro assessments were conducted using TLR2 knockout and both LLC-GFP and LN227 lentiviral-mediated knockdown (KD) cells. Results: Cancer cells released a secreted form of Hsp70 that acted on MΦ TLR2 to upregulate Mer receptor tyrosine kinase (MerTK) and induce MΦ M2 polarization. Hsp70 nAbs led to a reduction in CD14 expression by 75% in THP-1 cells in response to Gli36 EMD-CM. In addition, neutralizing TLR2 nAbs resulted in a 30% and 50% reduction in CD14 expression on THP-1 cells in response to MiaPaCa-2 and Gli36 exosome/microparticle-depleted conditioned media (EMD-CMs), respectively. Hsp70, TLR2, and MerTK formed a protein complex. Tumor growth and intra-tumor M2 MΦs were significantly reduced upon cancer cell Hsp70 knockdown and in TLR2 knockout mice. Conclusions: Cancer-secreted Hsp70 interacts with TLR2, upregulates MerTK on MΦs, and induces immunosuppressive MΦ M2 polarization. This previously unreported action of secreted Hsp70 suggests that disrupting the Hsp70-TLR2-MerTK interaction could serve as a promising immunotherapeutic approach to mitigate TME immunosuppression in solid cancers.

Dataset Information

Tumor-released autophagosomes induces CD4⁺ T cell-mediated immunosuppression via a TLR2-IL-6 cascade.

Background

Methods

Results

Conclusions

Publications

Tumor-released autophagosomes induces CD4<sup>+</sup> T cell-mediated immunosuppression via a TLR2-IL-6 cascade.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Dataset Information

Tumor-released autophagosomes induces CD4+ T cell-mediated immunosuppression via a TLR2-IL-6 cascade.

Background

Methods

Results

Conclusions

Publications

Tumor-released autophagosomes induces CD4<sup>+</sup> T cell-mediated immunosuppression via a TLR2-IL-6 cascade.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Tumor-released autophagosomes induces CD4⁺ T cell-mediated immunosuppression via a TLR2-IL-6 cascade.