p66Shc deficiency in CLL cells enhances PD-L1 expression and suppresses immune synapse formation
Introduction: Escape from immunosurveillance is a hallmark of chronic lymphocytic leukemia (CLL) cells. In the protective niche of lymphoid organs, leukemic cells suppress the ability of T lymphocytes to form the immune synapse (IS), thereby hampering T-cell mediated anti-tumoral activities. By bind...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2024-01-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2024.1297116/full |
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| author | Ludovica Lopresti Nagaja Capitani Vanessa Tatangelo Carmela Tangredi Gioia Boncompagni Federica Frezzato Andrea Visentin Giuseppe Marotta Sara Ciofini Alessandro Gozzetti Monica Bocchia Livio Trentin Cosima T. Baldari Laura Patrussi |
| author_facet | Ludovica Lopresti Nagaja Capitani Vanessa Tatangelo Carmela Tangredi Gioia Boncompagni Federica Frezzato Andrea Visentin Giuseppe Marotta Sara Ciofini Alessandro Gozzetti Monica Bocchia Livio Trentin Cosima T. Baldari Laura Patrussi |
| author_sort | Ludovica Lopresti |
| collection | DOAJ |
| description | Introduction: Escape from immunosurveillance is a hallmark of chronic lymphocytic leukemia (CLL) cells. In the protective niche of lymphoid organs, leukemic cells suppress the ability of T lymphocytes to form the immune synapse (IS), thereby hampering T-cell mediated anti-tumoral activities. By binding its cognate receptor PD-1 at the surface of T lymphocytes, the inhibitory ligand PD-L1, which is overexpressed in CLL cells, mediates the T-cell suppressive activities of CLL cells. However, the molecular mechanism underlying PD-L1 overexpression in CLL cells remains unknown. We have previously reported a defective expression of the pro-apoptotic and pro-oxidant adaptor p66Shc in CLL cells, which is causally related to an impairment in intracellular reactive oxygen species (ROS) production and to the activation of the ROS-sensitive transcription factor NF-κB. The fact that PD-L1 expression is regulated by NF-κB suggests a mechanistic relationship between p66Shc deficiency and PD-L1 overexpression in CLL cells.Methods: 62 treatment-naive CLL patients and 43 healthy donors were included in this study. PD-L1 and p66Shc expression was quantified in B cells by flow cytometry and qRT-PCR. IS architecture and local signaling was assessed by flow cytometry and confocal microscopy. CD8+ cell killing activity was assessed by flow cytometry.Results: Here we show that residual p66Shc expression in leukemic cells isolated both from CLL patients and from the CLL mouse model Eμ-TCL1 inversely correlated with PD-L1 expression. We also show that the PD-L1 increase prevented leukemic cells from forming ISs with T lymphocytes. Reconstitution of p66Shc, but not of a ROS-defective mutant, in both CLL cells and the CLL-derived cell line MEC-1, enhanced intracellular ROS and decreased PD-L1 expression. Similar results were obtained following treatment of CLL cells with H2O2 as exogenous source of ROS, that normalized PD-L1 expression and recovered IS formation.Discussion: Our data provide direct evidence that the p66Shc-deficiency-related ROS depletion in CLL cells concurs to enhance PD-L1 expression and provides a mechanistic basis for the suppression of T cell-mediated anti-tumoral functions in the immunosuppressive lymphoid niche. |
| first_indexed | 2024-03-08T04:13:06Z |
| format | Article |
| id | doaj.art-e7e9e6a473054becbee4aeecfc465039 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-03-08T04:13:06Z |
| publishDate | 2024-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-e7e9e6a473054becbee4aeecfc4650392024-02-08T16:52:37ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2024-01-011210.3389/fcell.2024.12971161297116p66Shc deficiency in CLL cells enhances PD-L1 expression and suppresses immune synapse formationLudovica Lopresti0Nagaja Capitani1Vanessa Tatangelo2Carmela Tangredi3Gioia Boncompagni4Federica Frezzato5Andrea Visentin6Giuseppe Marotta7Sara Ciofini8Alessandro Gozzetti9Monica Bocchia10Livio Trentin11Cosima T. Baldari12Laura Patrussi13Department of Life Sciences, University of Siena, Siena, ItalyDepartment of Life Sciences, University of Siena, Siena, ItalyDepartment of Life Sciences, University of Siena, Siena, ItalyDepartment of Life Sciences, University of Siena, Siena, ItalyDepartment of Life Sciences, University of Siena, Siena, ItalyHematology and Clinical Immunology Unit, Department of Medicine, University of Padua, Padua, ItalyHematology and Clinical Immunology Unit, Department of Medicine, University of Padua, Padua, ItalyStem Cell Transplant and Cellular Therapy Unit, University Hospital, Siena, ItalyDepartment of Medical Science, Surgery and Neuroscience, University of Siena, Siena, ItalyDepartment of Medical Science, Surgery and Neuroscience, University of Siena, Siena, ItalyDepartment of Medical Science, Surgery and Neuroscience, University of Siena, Siena, ItalyHematology and Clinical Immunology Unit, Department of Medicine, University of Padua, Padua, ItalyDepartment of Life Sciences, University of Siena, Siena, ItalyDepartment of Life Sciences, University of Siena, Siena, ItalyIntroduction: Escape from immunosurveillance is a hallmark of chronic lymphocytic leukemia (CLL) cells. In the protective niche of lymphoid organs, leukemic cells suppress the ability of T lymphocytes to form the immune synapse (IS), thereby hampering T-cell mediated anti-tumoral activities. By binding its cognate receptor PD-1 at the surface of T lymphocytes, the inhibitory ligand PD-L1, which is overexpressed in CLL cells, mediates the T-cell suppressive activities of CLL cells. However, the molecular mechanism underlying PD-L1 overexpression in CLL cells remains unknown. We have previously reported a defective expression of the pro-apoptotic and pro-oxidant adaptor p66Shc in CLL cells, which is causally related to an impairment in intracellular reactive oxygen species (ROS) production and to the activation of the ROS-sensitive transcription factor NF-κB. The fact that PD-L1 expression is regulated by NF-κB suggests a mechanistic relationship between p66Shc deficiency and PD-L1 overexpression in CLL cells.Methods: 62 treatment-naive CLL patients and 43 healthy donors were included in this study. PD-L1 and p66Shc expression was quantified in B cells by flow cytometry and qRT-PCR. IS architecture and local signaling was assessed by flow cytometry and confocal microscopy. CD8+ cell killing activity was assessed by flow cytometry.Results: Here we show that residual p66Shc expression in leukemic cells isolated both from CLL patients and from the CLL mouse model Eμ-TCL1 inversely correlated with PD-L1 expression. We also show that the PD-L1 increase prevented leukemic cells from forming ISs with T lymphocytes. Reconstitution of p66Shc, but not of a ROS-defective mutant, in both CLL cells and the CLL-derived cell line MEC-1, enhanced intracellular ROS and decreased PD-L1 expression. Similar results were obtained following treatment of CLL cells with H2O2 as exogenous source of ROS, that normalized PD-L1 expression and recovered IS formation.Discussion: Our data provide direct evidence that the p66Shc-deficiency-related ROS depletion in CLL cells concurs to enhance PD-L1 expression and provides a mechanistic basis for the suppression of T cell-mediated anti-tumoral functions in the immunosuppressive lymphoid niche.https://www.frontiersin.org/articles/10.3389/fcell.2024.1297116/fullPD-L1immune checkpointCLLp66Shcimmune synapseROS |
| spellingShingle | Ludovica Lopresti Nagaja Capitani Vanessa Tatangelo Carmela Tangredi Gioia Boncompagni Federica Frezzato Andrea Visentin Giuseppe Marotta Sara Ciofini Alessandro Gozzetti Monica Bocchia Livio Trentin Cosima T. Baldari Laura Patrussi p66Shc deficiency in CLL cells enhances PD-L1 expression and suppresses immune synapse formation Frontiers in Cell and Developmental Biology PD-L1 immune checkpoint CLL p66Shc immune synapse ROS |
| title | p66Shc deficiency in CLL cells enhances PD-L1 expression and suppresses immune synapse formation |
| title_full | p66Shc deficiency in CLL cells enhances PD-L1 expression and suppresses immune synapse formation |
| title_fullStr | p66Shc deficiency in CLL cells enhances PD-L1 expression and suppresses immune synapse formation |
| title_full_unstemmed | p66Shc deficiency in CLL cells enhances PD-L1 expression and suppresses immune synapse formation |
| title_short | p66Shc deficiency in CLL cells enhances PD-L1 expression and suppresses immune synapse formation |
| title_sort | p66shc deficiency in cll cells enhances pd l1 expression and suppresses immune synapse formation |
| topic | PD-L1 immune checkpoint CLL p66Shc immune synapse ROS |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2024.1297116/full |
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