Abstract
Patients with non-small-cell lung cancer (NSCLC) have immune defects that are poorly understood. Forkhead box protein P3 (Foxp3) is crucial for immunosuppression by CD4+ regulatory T cells (Tregs). It is not well known how NSCLC induces Foxp3 expression and causes immunosuppression in tumor-bearing patients. Our study found a higher percentage of CD4+ Tregs in the peripheral blood of NSCLC compared with healthy donors. NSCLC patients showed demethylation of eight CpG sites within the Foxp3 promoter with methylation ratios negatively correlated with CD4+CD25+Foxp3+ T levels. Foxp3 expression in CD4+ Tregs was directly regulated by Foxp3 promoter demethylation and was involved in immunosuppression by NSCLC. To verify the effect of tumor cells on the phenotype and function of CD4+ Tregs, we established a coculture system using NSCLC cell line and healthy CD4+ T cells and showed that SPC-A1 induced IL-10 and TGF-β1 secretion by affecting the function of CD4+ Tregs. The activity of DNA methyltransferases from CD4+ T was decreased during this process. Furthermore, eight CpG sites within the Foxp3 promoter also appeared to have undergone demethylation. Foxp3 is highly expressed in CD4+ T cells, and this may be caused by gene promoter demethylation. These induced Tregs are highly immunosuppressive and dramatically inhibit the proliferative activity of naïve CD4+ T cells. Our study provides one possible mechanism describing Foxp3 promoter demethylation changes by which NSCLC down-regulates immune responses and contributes to tumor progression. Foxp3 represents an important target for NSCLC anti-tumor immunotherapy.
Abbreviations
- AP-1:
-
Activator protein-1
- DNMTs:
-
DNA methyltransferases
- ELISA:
-
Enzyme-linked immunosorbent assay
- Foxp3:
-
Forkhead box protein P3
- IL:
-
Interleukin
- NF-AT:
-
Nuclear factor of activated T cells
- NSCLC:
-
Non-small-cell lung cancer
- PBMCs:
-
Peripheral blood mononuclear cells
- SAM:
-
S-adenosylmethionine
- SCC:
-
Squamous cell carcinoma
- TGF:
-
Transforming growth factor
- Tregs:
-
Regulatory T cells
- TSDR:
-
Treg-specific demethylated region
- TSS:
-
Transcriptional start site
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Acknowledgments
We are grateful to the technical support from National Key Clinical Department of Laboratory Medicine of Jiangsu Province Hospital. This work was supported by National Natural Science Foundation of China (Nos. 81272324, 81371894) and Key Laboratory for Medicine of Jiangsu Province of China (No. XK201114), a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.
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Xing Ke and Shuping Zhang have contributed equally to this work.
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Ke, X., Zhang, S., Xu, J. et al. Non-small-cell lung cancer-induced immunosuppression by increased human regulatory T cells via Foxp3 promoter demethylation. Cancer Immunol Immunother 65, 587–599 (2016). https://doi.org/10.1007/s00262-016-1825-6
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DOI: https://doi.org/10.1007/s00262-016-1825-6