Raffaele De Palma1, Plinio Cirillo2, Giovanni Ciccarelli3, Giusi Barra4, Stefano Conte3, Grazia Pellegrino2, Giuseppe Pasquale4, Giovanni Nassa5, Francesco Pacifico6, Antonio Leonardi7, Luigi Insabato2, Gaetano Calì8, Paolo Golino9, Giovanni Cimmino3. 1. Department of Clinical and Experimental Medicine, Section of Clinical Immunology, Second University of Naples, Naples, Italy; Institute of Protein Biochemistry, CNR, Naples, Italy. 2. Department of Advanced Biosciences, Section of Cardiology, University of Naples, "Federico II", Naples, Italy. 3. Department of Cardiothoracic and Respiratory Sciences, Section of Cardiology, Second University of Naples, Naples, Italy. 4. Department of Clinical and Experimental Medicine, Section of Clinical Immunology, Second University of Naples, Naples, Italy. 5. Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, Baronissi, SA, Italy. 6. Endocrinology and Experimental Oncology Institute, CNR, Naples, Italy. 7. Department of Molecular and Cellular Biology and Pathology, University of Naples, "Federico II", Naples, Italy. 8. Endocrinology and Experimental Oncology Institute, CNR, Naples, Italy; Department of Molecular Medicine and Medical Biotechnologies, University "Federico II", Naples, Italy. 9. Department of Cardiothoracic and Respiratory Sciences, Section of Cardiology, Second University of Naples, Naples, Italy. Electronic address: paolo.golino@unina2.it.
Abstract
OBJECTIVE: T-lymphocyte activation plays an important role in the pathophysiology of acute coronary syndromes (ACS). Plaques from ACS patients show a selective oligoclonal expansion of T-cells, indicating a specific, antigen-driven recruitment of T-lymphocytes within the unstable lesions. At present, however, it is not known whether T-cells may contribute directly to thrombosis by expressing functional tissue factor (TF). Accordingly, the aim of the present study was to investigate whether T-cells are able to express functional TF in their activated status. METHODS: In vitro, CD3(+)-cells, isolated from buffy coats, were stimulated with anti-CD3/CD28 beads, IL-6, TNF-α, IL-17, INF-γ or PMA/ionomycin. Following stimulation, TF expression on cell-surface, at gene and protein levels, as well as its procoagulant activity in whole cells and microparticles was measured. In vivo, TF expression was evaluated in CD3(+)-cells isolated from the aorta and the coronary sinus of ACS-NSTEMI and stable coronary artery disease (SCAD) patients. The presence of CD3(+)-TF(+)cells was also evaluated by immunohistochemistry in thrombi aspirated from ACS-STEMI patients. RESULTS: PMA/ionomycin and IL-17 plus INF-γ stimulation resulted in a significant TF increase at gene and protein levels as well as at cell-surface expression. This was accompanied by a parallel increase in FXa generation, both in whole cells and in microparticles, indicating that the induced membrane-bound TF was active. Furthermore, transcardiac TF gradient was significantly higher in CD3(+)-cells obtained from ACS-patients compared to SCAD-patients. Interestingly, thrombi from ACS-STEMI patients resulted enriched in CD3(+)-cells, most of them expressing TF. CONCLUSIONS: Our data demonstrate that activated T-lymphocytes in vitro express functional TF on their membranes, suggesting a direct pathophysiological role of these cells in the thrombotic process; this hypothesis is further supported by the observations in vivo that CD3(+)-cells from coronary circulation of ACS-NSTEMI patients show increased TF levels and that coronary thrombi from ACS-STEMI patients are enriched in CD3(+)-cells expressing TF.
OBJECTIVE: T-lymphocyte activation plays an important role in the pathophysiology of acute coronary syndromes (ACS). Plaques from ACS patients show a selective oligoclonal expansion of T-cells, indicating a specific, antigen-driven recruitment of T-lymphocytes within the unstable lesions. At present, however, it is not known whether T-cells may contribute directly to thrombosis by expressing functional tissue factor (TF). Accordingly, the aim of the present study was to investigate whether T-cells are able to express functional TF in their activated status. METHODS: In vitro, CD3(+)-cells, isolated from buffy coats, were stimulated with anti-CD3/CD28 beads, IL-6, TNF-α, IL-17, INF-γ or PMA/ionomycin. Following stimulation, TF expression on cell-surface, at gene and protein levels, as well as its procoagulant activity in whole cells and microparticles was measured. In vivo, TF expression was evaluated in CD3(+)-cells isolated from the aorta and the coronary sinus of ACS-NSTEMI and stable coronary artery disease (SCAD) patients. The presence of CD3(+)-TF(+)cells was also evaluated by immunohistochemistry in thrombi aspirated from ACS-STEMI patients. RESULTS:PMA/ionomycin and IL-17 plus INF-γ stimulation resulted in a significant TF increase at gene and protein levels as well as at cell-surface expression. This was accompanied by a parallel increase in FXa generation, both in whole cells and in microparticles, indicating that the induced membrane-bound TF was active. Furthermore, transcardiac TF gradient was significantly higher in CD3(+)-cells obtained from ACS-patients compared to SCAD-patients. Interestingly, thrombi from ACS-STEMI patients resulted enriched in CD3(+)-cells, most of them expressing TF. CONCLUSIONS: Our data demonstrate that activated T-lymphocytes in vitro express functional TF on their membranes, suggesting a direct pathophysiological role of these cells in the thrombotic process; this hypothesis is further supported by the observations in vivo that CD3(+)-cells from coronary circulation of ACS-NSTEMI patients show increased TF levels and that coronary thrombi from ACS-STEMI patients are enriched in CD3(+)-cells expressing TF.
Authors: Pegah Mir Seyed Nazari; Anna S Berghoff; Matthias Preusser; Florian Moik; Florian Posch; Gerda Ricken; Julia Riedl; Lena Hell; Christine Marosi; Johannes A Hainfellner; Ingrid Pabinger; Cihan Ay Journal: ESMO Open Date: 2020-05
Authors: Mingqing Song; Zachary W Fitch; Kannan P Samy; Benjamin M Martin; Qimeng Gao; Robert Patrick Davis; Francis V Leopardi; Niki Huffman; Robin Schmitz; Gayathri R Devi; Bradley H Collins; Allan D Kirk Journal: Xenotransplantation Date: 2021-02-22 Impact factor: 3.907
Authors: Giovanni Cimmino; Francesco S Loffredo; Alberto Morello; Saverio D'Elia; Raffaele De Palma; Plinio Cirillo; Paolo Golino Journal: Curr Cardiol Rev Date: 2017