Breast Cancer Cell-Derived Adenosine Enhances Generation and Suppressor Function of Human Adaptive Regulatory T Cells
Introduction: Adaptive regulatory T cells (Tr1) are induced in the periphery by environmental stimuli. CD73 expression and adenosine (ADO) production by tumor cells may influence Tr1 generation and their immunosuppressive activity. Material and Methods: Tr1 were generated in co-cultures of CD4+CD25n...
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MDPI AG
2021-07-01
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| Series: | Journal of Personalized Medicine |
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| Online Access: | https://www.mdpi.com/2075-4426/11/8/754 |
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| author | Magis Mandapathil Miroslaw J. Szczepanski Edwin K. Jackson Stephan Lang Theresa L. Whiteside |
| author_facet | Magis Mandapathil Miroslaw J. Szczepanski Edwin K. Jackson Stephan Lang Theresa L. Whiteside |
| author_sort | Magis Mandapathil |
| collection | DOAJ |
| description | Introduction: Adaptive regulatory T cells (Tr1) are induced in the periphery by environmental stimuli. CD73 expression and adenosine (ADO) production by tumor cells may influence Tr1 generation and their immunosuppressive activity. Material and Methods: Tr1 were generated in co-cultures of CD4+CD25neg T cells, autologous immature dendritic cells (iDC), and irradiated ADO-producing CD73+ or non-producing CD73neg breast cancer (BrCa) cell lines (TU). The expression of ectonucleotidases and other surface markers on Tr1 was determined by flow cytometry. Tr1-mediated suppression of proliferation was evaluated in CFSE-based assays. Luciferase-based ATP detection assays and mass spectrometry were used to measure ATP hydrolysis and ADO levels. Cytokine levels were measured by ELISA or Luminex. CD73 expression on tumor cells or T cells in TU tissues was assessed by immunofluorescence. Results: CD73+ TU induced higher numbers of Tr1 cells (<i>p</i> < 0.01) than CD73neg TU. Tr1TU73+ hydrolyzed more exogenous ATP, produced more ADO, and mediated higher suppression than Tr1TU73neg (<i>p</i> < 0.05 for all). ARL67156, an ectonucleotidase inhibitor, and ZM241385, A2A receptor antagonist, reduced suppression of proliferation mediated by Tr1TU73+ cells (<i>p</i> < 0.01). Basal-like primary BrCa cells expressed higher levels of ectonucleotidases and induced more Tr1 than less aggressive primary luminal-like BrCa. Conclusion: BrCa producing ADO (CD73+ TU) favor the induction of Tr1, which expresses CD39 and CD73, hydrolyzes ATP to ADO, and effectively suppresses anti-tumor immunity. |
| first_indexed | 2024-03-10T08:40:43Z |
| format | Article |
| id | doaj.art-2619cdf8724d40efa80e8d878f75b4ab |
| institution | Directory Open Access Journal |
| issn | 2075-4426 |
| language | English |
| last_indexed | 2024-03-10T08:40:43Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Journal of Personalized Medicine |
| spelling | doaj.art-2619cdf8724d40efa80e8d878f75b4ab2023-11-22T08:18:51ZengMDPI AGJournal of Personalized Medicine2075-44262021-07-0111875410.3390/jpm11080754Breast Cancer Cell-Derived Adenosine Enhances Generation and Suppressor Function of Human Adaptive Regulatory T CellsMagis Mandapathil0Miroslaw J. Szczepanski1Edwin K. Jackson2Stephan Lang3Theresa L. Whiteside4Department of Otorhinolaryngology, Asklepios Clinic St. Georg, 20099 Hamburg, GermanyDepartment of Biochemistry, Medical University of Warsaw, PL-02097 Warsaw, PolandDepartment of Pharmacology, University of Pittsburgh, Pittsburgh, PA 15219, USADepartment of Otorhinolaryngology, University of Duisburg-Essen, 45147 Essen, GermanyUPMC Hillman Cancer Center, Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USAIntroduction: Adaptive regulatory T cells (Tr1) are induced in the periphery by environmental stimuli. CD73 expression and adenosine (ADO) production by tumor cells may influence Tr1 generation and their immunosuppressive activity. Material and Methods: Tr1 were generated in co-cultures of CD4+CD25neg T cells, autologous immature dendritic cells (iDC), and irradiated ADO-producing CD73+ or non-producing CD73neg breast cancer (BrCa) cell lines (TU). The expression of ectonucleotidases and other surface markers on Tr1 was determined by flow cytometry. Tr1-mediated suppression of proliferation was evaluated in CFSE-based assays. Luciferase-based ATP detection assays and mass spectrometry were used to measure ATP hydrolysis and ADO levels. Cytokine levels were measured by ELISA or Luminex. CD73 expression on tumor cells or T cells in TU tissues was assessed by immunofluorescence. Results: CD73+ TU induced higher numbers of Tr1 cells (<i>p</i> < 0.01) than CD73neg TU. Tr1TU73+ hydrolyzed more exogenous ATP, produced more ADO, and mediated higher suppression than Tr1TU73neg (<i>p</i> < 0.05 for all). ARL67156, an ectonucleotidase inhibitor, and ZM241385, A2A receptor antagonist, reduced suppression of proliferation mediated by Tr1TU73+ cells (<i>p</i> < 0.01). Basal-like primary BrCa cells expressed higher levels of ectonucleotidases and induced more Tr1 than less aggressive primary luminal-like BrCa. Conclusion: BrCa producing ADO (CD73+ TU) favor the induction of Tr1, which expresses CD39 and CD73, hydrolyzes ATP to ADO, and effectively suppresses anti-tumor immunity.https://www.mdpi.com/2075-4426/11/8/754TregadenosineCD73breast cancer (BrCa)immunosuppression |
| spellingShingle | Magis Mandapathil Miroslaw J. Szczepanski Edwin K. Jackson Stephan Lang Theresa L. Whiteside Breast Cancer Cell-Derived Adenosine Enhances Generation and Suppressor Function of Human Adaptive Regulatory T Cells Journal of Personalized Medicine Treg adenosine CD73 breast cancer (BrCa) immunosuppression |
| title | Breast Cancer Cell-Derived Adenosine Enhances Generation and Suppressor Function of Human Adaptive Regulatory T Cells |
| title_full | Breast Cancer Cell-Derived Adenosine Enhances Generation and Suppressor Function of Human Adaptive Regulatory T Cells |
| title_fullStr | Breast Cancer Cell-Derived Adenosine Enhances Generation and Suppressor Function of Human Adaptive Regulatory T Cells |
| title_full_unstemmed | Breast Cancer Cell-Derived Adenosine Enhances Generation and Suppressor Function of Human Adaptive Regulatory T Cells |
| title_short | Breast Cancer Cell-Derived Adenosine Enhances Generation and Suppressor Function of Human Adaptive Regulatory T Cells |
| title_sort | breast cancer cell derived adenosine enhances generation and suppressor function of human adaptive regulatory t cells |
| topic | Treg adenosine CD73 breast cancer (BrCa) immunosuppression |
| url | https://www.mdpi.com/2075-4426/11/8/754 |
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