182 results on '"Antonio Tabilio"'
Search Results
2. Data from Endothelial Differentiation of Hematopoietic Stem and Progenitor Cells from Patients with Multiple Myeloma
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Angelo Vacca, Franco Dammacco, Domenico Ribatti, Attilio Guarini, Nicola Di Renzo, Antonio Tabilio, Diego Guidolin, Giuseppe Mangialardi, Franca Falzetti, Teresa Cirulli, Claudia Piccoli, and Roberto Ria
- Abstract
Purpose: Vasculogenesis is a physiologic process typical of fetal development in which new blood vessels develop from undifferentiated precursors (or angioblasts). In tumors, near angiogenesis, vasculogenesis contributes to the formation of the microvascular plexus that is important for diffusion. Here, we show that hematopoietic stem and progenitor cells (HSPC) of multiple myeloma (MM) patients are able to differentiate into cells with endothelial phenotype on exposure to angiogenic cytokines.Experimental Design: Circulating HSPCs were purified with an anti-CD133 antibody from patients with newly diagnosed MM before autologous transplantation and exposed to vascular endothelial growth factor (VEGF), fibroblast growth factor-2 and insulin-like growth factor in a 3-week culture.Results: HSPCs gradually lost CD133 expression and acquired VEGF receptor-2, factor VIII–related antigen, and vascular endothelial-cadherin expression. The expression pattern overlapped with paired MM endothelial cells (MMEC). During culture, cells adhered to fibronectin, spread, and acquired an endothelial cell shape. Differentiated HSPCs also became capillarogenic in the Matrigel assay with maximal activity at the third week of culture. Bone marrow biopsies revealed HSPCs inside the neovessel wall in patients with MM but not in those with monoclonal gammopathy of undetermined significance.Conclusions: In patients with MM, but not in those with monoclonal gammopathy of undetermined significance, HSPCs contribute to the neovessel wall building together with MMECs. Therefore, besides angiogenesis, HSPC-linked vasculogenesis contributes to neovascularization in MM patients. Tentatively, we hypothesize that in HSPC cultures a multipotent cell population expressing low VEGF receptor-2 levels corresponds to the endothelial progenitor cell precursor and seems to be the MMEC precursor.
- Published
- 2023
3. Activated autologous T cells exert an anti-B-cell chronic lymphatic leukemia effect in vitro and in vivo
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Franca Falzetti, Raffaella Ciurnelli, Adelmo Terenzi, Emanuela Rosati, Alessandro Lucchesi, Federico Bazzucchi, Antonio Tabilio, Pier Francesco Marconi, Moira Bazzucchi, Mauro Di Ianni, Beatrice Del Papa, Lorenzo Moretti, and Paolo Sportoletti
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Cytotoxicity, Immunologic ,Cancer Research ,NOD/SCID mice ,T-Lymphocytes ,CD3 ,Immunology ,T lymphocytes ,Receptors, Antigen, T-Cell ,Down-Regulation ,Mice, SCID ,Biology ,Lymphocyte Activation ,Mice ,Immune system ,immune system diseases ,In vivo ,Cell Line, Tumor ,hemic and lymphatic diseases ,medicine ,Animals ,Humans ,Immunologic Factors ,Immunology and Allergy ,Gene Rearrangement, B-Lymphocyte ,Cytotoxicity ,Genetics (clinical) ,B cell ,Cell Proliferation ,Transplantation ,Gene Expression Profiling ,Cell Biology ,Autologous anti-tumoral activity ,chronic lymphatic leukemia ,gene expression profiling ,OKT3 activation ,Leukemia, Lymphocytic, Chronic, B-Cell ,Molecular biology ,In vitro ,Up-Regulation ,medicine.anatomical_structure ,Oncology ,Apoptosis ,biology.protein ,Interleukin-2 ,Somatic Hypermutation, Immunoglobulin ,Muromonab-CD3 ,K562 cells - Abstract
The impact of chronic lymphatic leukemia (CLL) tumor burden on the autologous immune system has already been demonstrated. This study attempted to elucidate the molecular mechanisms underlying T-cell immunologic deficiencies in CLL.Freshly isolated CD3(+) T cells from patients with a diagnosis of CLL and healthy donors were analyzed by gene expression profiling. Activated T cells from 20 patients with CLL were tested in vitro for cytotoxicity against mutated and unmutated autologous B cells and DAUDI, K562 and P815 cell lines. To investigate T-cell mediated cytotoxicity in vivo, we co-transplanted OKT3-activated T lymphocytes and autologous B-cell CLL (B-CLL) cells into NOD/SCID mice.Gene expression profiles of peripheral blood T cells from B-CLL patients showed 25 down-regulated, and 31 up-regulated, genes that were mainly involved in cell differentiation, proliferation, survival, apoptosis, cytoskeleton formation, vesicle trafficking and T-cell activation. After culture, the T-cell count remained unchanged, CD8 cells expanded more than CD4 and a cytotoxicity index30% was present in 5/20 patients. Cytotoxicity against B autologous leukemic cells did not correlate with B-cell mutational status. Only activated T cells exerting cytotoxicity against autologous leukemic B cells prevented CLL in a human-mouse chimera.This study indicates that patients with CLL are affected by a partial immunologic defect that might be somewhat susceptible to repair. This study identifies the molecular pathways underlying T-cell deficiencies in CLL and shows that cytotoxic T-cell functions against autologous B-CLL can be rebuilt at least in part in vitro and in vivo.
- Published
- 2009
4. Toxic epidermal necrolysis in a patient with primary myelofibrosis receiving thalidomide therapy
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Marianna Colagrande, Antonio Tabilio, Lorenzo Moretti, Gino Coletti, Maria Concetta Fargnoli, Mario Lapecorella, Mauro Di Ianni, and Ketty Peris
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Male ,medicine.medical_specialty ,Erythroderma ,Internal medicine ,Humans ,Medicine ,Myelofibrosis ,Aged ,Hematology ,Neovascularization, Pathologic ,business.industry ,medicine.disease ,Dermatology ,Toxic epidermal necrolysis ,Thalidomide ,Surgery ,Extramedullary hematopoiesis ,Transplantation ,medicine.anatomical_structure ,Primary Myelofibrosis ,Stevens-Johnson Syndrome ,Bone marrow ,business ,medicine.drug - Abstract
Primary myelofibrosis (PMF) is a chronic myeloproliferative neoplasm characterized by progressive anemia, massive splenomegaly, leukoerythroblastosis, extramedullary hematopoiesis and in about 50% of cases the presence of JAK2V617F mutation. Curative therapy in PMF is currently possible only with allogeneic haematopoietic stem cell transplantation which is, unfortunately, associated with relatively high risks of mortality and morbidity which undermine its broad applications. Non-transplant treatment modalities are used for palliative purposes. Recently, anti-angiogenic drugs such as thalidomide have been used to treat these patients on the basis of the prominent bone marrow angiogenesis. Here, we report the case of a patient suffering from JAK2V617F-positive PMF with marked bone marrow neo-angiogenesis. The patient was treated with thalidomide but after 20 days developed life-threatening toxic epidermal necrolysis (TEN). To the best of our knowledge this is the first case of TEN in a patient with PMF under thalidomide therapy.
- Published
- 2008
5. MtDNA mutation associated with mitochondrial dysfunction in megakaryoblastic leukaemic cells
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Lorenzo Moretti, Antonio Tabilio, P Stanziale, Domenico Boffoli, Claudia Piccoli, Rosella Scrima, B Biscottini, Massimo Carella, Maria Ripoli, M Di Ianni, and Nazzareno Capitanio
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Cancer Research ,medicine.medical_specialty ,Mitochondrial DNA ,Oxidative phosphorylation ,Mitochondrion ,Biology ,medicine.disease_cause ,DNA, Mitochondrial ,Oxidative Phosphorylation ,Oxygen Consumption ,Leukemia, Megakaryoblastic, Acute ,Internal medicine ,medicine ,Humans ,Genetics ,Mutation ,Hematology ,Cancer ,NADH Dehydrogenase ,medicine.disease ,Mitochondria ,Leukemia ,medicine.anatomical_structure ,Oncology ,Megakaryoblast ,Cancer research ,Reactive Oxygen Species - Abstract
MtDNA mutation associated with mitochondrial dysfunction in megakaryoblastic leukaemic cells
- Published
- 2008
6. Endothelial Differentiation of Hematopoietic Stem and Progenitor Cells from Patients with Multiple Myeloma
- Author
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Attilio Guarini, Claudia Piccoli, Diego Guidolin, Franca Falzetti, Giuseppe Mangialardi, Antonio Tabilio, Franco Dammacco, Domenico Ribatti, Nicola Di Renzo, Teresa Cirulli, Angelo Vacca, and Roberto Ria
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Cancer Research ,Pathology ,medicine.medical_specialty ,Angiogenesis ,Biopsy ,Neovascularization, Physiologic ,Biology ,Endothelial progenitor cell ,chemistry.chemical_compound ,Vasculogenesis ,Bone Marrow ,medicine ,Humans ,Progenitor cell ,Cells, Cultured ,Matrigel ,Cell Differentiation ,Hematopoietic Stem Cells ,Vascular endothelial growth factor ,Endothelial stem cell ,Adult Stem Cells ,Phenotype ,Oncology ,chemistry ,Endothelium, Vascular ,Stem cell ,Multiple Myeloma - Abstract
Purpose: Vasculogenesis is a physiologic process typical of fetal development in which new blood vessels develop from undifferentiated precursors (or angioblasts). In tumors, near angiogenesis, vasculogenesis contributes to the formation of the microvascular plexus that is important for diffusion. Here, we show that hematopoietic stem and progenitor cells (HSPC) of multiple myeloma (MM) patients are able to differentiate into cells with endothelial phenotype on exposure to angiogenic cytokines. Experimental Design: Circulating HSPCs were purified with an anti-CD133 antibody from patients with newly diagnosed MM before autologous transplantation and exposed to vascular endothelial growth factor (VEGF), fibroblast growth factor-2 and insulin-like growth factor in a 3-week culture. Results: HSPCs gradually lost CD133 expression and acquired VEGF receptor-2, factor VIII–related antigen, and vascular endothelial-cadherin expression. The expression pattern overlapped with paired MM endothelial cells (MMEC). During culture, cells adhered to fibronectin, spread, and acquired an endothelial cell shape. Differentiated HSPCs also became capillarogenic in the Matrigel assay with maximal activity at the third week of culture. Bone marrow biopsies revealed HSPCs inside the neovessel wall in patients with MM but not in those with monoclonal gammopathy of undetermined significance. Conclusions: In patients with MM, but not in those with monoclonal gammopathy of undetermined significance, HSPCs contribute to the neovessel wall building together with MMECs. Therefore, besides angiogenesis, HSPC-linked vasculogenesis contributes to neovascularization in MM patients. Tentatively, we hypothesize that in HSPC cultures a multipotent cell population expressing low VEGF receptor-2 levels corresponds to the endothelial progenitor cell precursor and seems to be the MMEC precursor.
- Published
- 2008
7. Eubacterium plautii infection in a kidney transplant recipient: a noteworthy case of pleural effusion and fever
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Antonio Tabilio, Francesco Pisani, Pierpaolo Di Cocco, L. Bonanni, Paola Mastrantonio, Giuseppe Orlando, M. D'Angelo, and Antonio Famulari
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Transplantation ,medicine.medical_specialty ,Pleural effusion ,Opportunistic infection ,business.industry ,Perforation (oil well) ,Respiratory disease ,medicine.disease ,Surgery ,Pleural disease ,Pleurisy ,medicine ,Antibiotic prophylaxis ,business ,Kidney transplantation - Abstract
We report a noteworthy case of Eubacterium plautii infection after kidney transplantation. Our 33-yr-old transplant recipient received standard care; his post-transplant course was uneventful. However, on day 44 he underwent an emergency laparotomy for perforation of the ileum. He was initially treated with ceftazidime, fluconazole and metronidazole, but his fever persisted, so he was switched to meropenem and vancocin. We could not find any cause for his infection. On day 70, his temperature normalized. On day 75, he developed severe leukopenia (280 cell/mL). His cytomegalovirus-DNA test result was negative, so all immunosuppressants, except for prednisone, were stopped; instead, antibiotic prophylaxis was started, using caspofungin, trimethoprim-sulfamethoxazole and ciprofloxacin. On day 83, he underwent percutaneous drainage of massive left pleural effusion. We repeatedly cultured the pleural liquid, but it was not till three wk later that we were finally able to identify the causative organism. We hypothesize that the microorganism - which normally resides on the surface of the intestinal lumen - entered the bloodstream via bacterial translocation, eventually colonizing the pleurae. This translocation was favored by our patient poor clinical condition, his immunosuppressive treatment and his heavy antibiotherapy. Our experience highlights the need for wiser use of antibiotics in transplant recipients.
- Published
- 2008
8. Bone-marrow derived hematopoietic stem/progenitor cells express multiple isoforms of NADPH oxidase and produce constitutively reactive oxygen species
- Author
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Nazzareno Capitanio, Maria Ripoli, Domenico Boffoli, Annamaria D'Aprile, Rosella Scrima, Antonio Tabilio, Claudia Piccoli, and Lucia Lecce
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Morpholines ,Blotting, Western ,Biophysics ,Antigens, CD34 ,Bone Marrow Cells ,Biology ,Models, Biological ,Biochemistry ,Gene Expression Regulation, Enzymologic ,chemistry.chemical_compound ,Onium Compounds ,Glutathione Peroxidase GPX1 ,Humans ,Enzyme Inhibitors ,Progenitor cell ,Molecular Biology ,Glutathione Peroxidase ,Microscopy, Confocal ,NADPH oxidase ,Reverse Transcriptase Polymerase Chain Reaction ,Superoxide Dismutase ,Acetophenones ,NADPH Oxidases ,NOX4 ,Hydrogen Peroxide ,Cell Biology ,Catalase ,Flow Cytometry ,Hematopoietic Stem Cells ,Immunohistochemistry ,Cell biology ,Isoenzymes ,Endothelial stem cell ,Haematopoiesis ,chemistry ,Chromones ,NOX1 ,Apocynin ,cardiovascular system ,biology.protein ,Reactive Oxygen Species ,Adult stem cell - Abstract
Consolidated evidence highlights the importance of redox signalling in poising the balance between self-renewal and differentiation in adult stem cells. The present study shows that human hematopoietic stem/progenitor cells (HSCs) constitutively generate low levels of hydrogen peroxide whose production is inhibited by DPI, apocynin, catalase, and LY294002 and scarcely stimulated by PMA. Moreover, it is shown that HSCs express at the mRNA and protein levels the catalytic subunits of NOX1, NOX2, and NOX4 isoforms of the NADPH oxidase family along with the complete battery of the regulatory subunits p22, p40, p47, p67, rac1, rac2, NOXO1, and NOXA1 as well as the splicing variant NOX2s and that the three NOX isoforms are largely co-expressed in the same HSC. These findings are interpreted in terms of a positive feed-back mechanism of NOXs activation enabling a fine tuning of the ROS level to be possibly used in redox-mediated signalling for growth and differentiation of HSCs.
- Published
- 2007
9. Recurrent primary plasmacytoma of the eyelid with rapid regional metastasis
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L. Marcomigni, Roberto Ria, Antonio Tabilio, Paolo Sportoletti, Michele Cimminiello, and M Di Ianni
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Adult ,Male ,Immunofixation ,Cancer Research ,Pathology ,medicine.medical_specialty ,Biopsy ,Eyelid Neoplasms ,Metastasis ,medicine ,Humans ,Lymph node ,biology ,medicine.diagnostic_test ,business.industry ,Remission Induction ,Hematology ,medicine.disease ,Treatment Outcome ,medicine.anatomical_structure ,Oncology ,Lymphatic Metastasis ,biology.protein ,Plasmacytoma ,Bone marrow ,Eyelid ,Lymph ,Neoplasm Recurrence, Local ,business ,Follow-Up Studies - Abstract
In this case, originally reported as primary eyelid plasmacytoma, the tumor recurred on the same eyelid within 2 years of surgery. No plasma cell infiltration was observed at bone marrow biopsy. No serum or urinary monoclonal component was detected at immunofixation. Histology and immunohistochemistry confirmed plasma cell infiltration. Tumor cell clonality was determined by immunohistological staining; cells were positive for kappa light chain like the first eyelid tumor. Surgery was followed by radiotherapy. Twenty months later, biopsy of one enlarged right cervical lymph node showed massive diffuse infiltration of atypical plasma cells (CD20(-), CD79a(+), CD138(+), MUM1/IRF4(+)). Given the rapid diffusion to lymph nodes and the appearance of the monoclonal component, the lymph node was removed surgically. No adjuvant chemotherapy was given. Unexpectedly, the serum monoclonal component normalized. No plasma cell infiltration was observed at bone marrow biopsy. As this case might be a particularly slow-progressing extra-medullary plasmacytoma, this study recommends closely monitored follow-ups so that the aggressive form can be treated in time.
- Published
- 2006
10. Characterization of Mitochondrial and Extra-mitochondrial Oxygen Consuming Reactions in Human Hematopoietic Stem Cells
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Domenico Boffoli, Olga Cela, Rosella Scrima, Franca Falzetti, Claudia Piccoli, Antonio Tabilio, Nazzareno Capitanio, Annamaria D'Aprile, and Roberto Ria
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Oxidase test ,Hematopoietic stem cell ,Cell Biology ,NAD(P)H oxidase activity ,Mitochondrion ,Biology ,Biochemistry ,Cell biology ,Mitochondrial respiratory chain ,medicine.anatomical_structure ,medicine ,NAD+ kinase ,Stem cell ,Molecular Biology ,Intracellular - Abstract
This study was aimed to characterize the mitochondrial and extra-mitochondrial oxygen consuming reactions in human CD34+ hematopoietic stem cells. Cell samples were collected by apheresis following pre-conditioning by granulocyte colony-stimulating factor and isolated by anti-CD34 positive immunoselection. Polarographic analysis of the CN-sensitive endogenous cell respiration revealed a low mitochondrial oxygen consumption rate. Differential absorbance spectrometry on whole cell lysate and two-dimensional blue native-PAGE analysis of mitoplast proteins confirmed a low amount of mitochondrial respiratory chain complexes thus qualifying the hematopoietic stem cell as a poor oxidative phosphorylating cell type. Confocal microscopy imaging showed, however, that the intracellular content of mitochondria was not homogeneously distributed in the CD34+ hematopoietic stem cell sample displaying a clear inverse correlation of their density with the expression of the CD34 commitment marker. About half of the endogenous oxygen consumption was extra-mitochondrial and completely inhibitable by enzymatic scavengers of reactive oxygen species and by diphenylene iodinium. By spectral analysis, flow cytometry, reverse transcriptase-PCR, immunocytochemistry, and immunoprecipitation it was shown that the extra-mitochondrial oxygen consumption was contributed by the NOX2 and NOX4 isoforms of the O2-*. producer plasma membrane NAD(P)H oxidase with low constitutive activity. A model is proposed suggesting for the NAD(P)H oxidase a role of O2 sensor and/or ROS source serving as redox messengers in the activation of intracellular signaling pathways leading (or contributing) to mitochondriogenesis, cell survival, and differentiation in hematopoietic stem cells.
- Published
- 2005
11. Interleukin 7-Engineered Stromal Cells: A New Approach for Hastening Naive T Cell Recruitment
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Antonio Tabilio, Mauro Di Ianni, Maria De Ioanni, Tiziana Zei, Claude Bagnis, Lorenzo Moretti, Adelmo Terenzi, Massimo F. Martelli, Patrice Mannoni, Franca Falzetti, Paolo Sportoletti, Elisabetta Bonifacio, Brunangelo Falini, Beatrice Del Papa, Fabrizio Spinozzi, and Eugenia Gaozza
- Subjects
CD4-Positive T-Lymphocytes ,Stromal cell ,CD3 Complex ,Naive T cell ,Cell Transplantation ,T-Lymphocytes ,Antigens, CD34 ,Mice, SCID ,CD8-Positive T-Lymphocytes ,Biology ,Cell Line ,Mice ,Interleukin 21 ,Mice, Inbred NOD ,Transduction, Genetic ,Genetics ,Lymph node stromal cell ,Animals ,Humans ,Cytotoxic T cell ,Molecular Biology ,Interleukin 3 ,Interleukin-7 ,Interleukin ,Retroviridae ,Immunology ,Interleukin 12 ,Cancer research ,Leukocyte Common Antigens ,Molecular Medicine ,Stromal Cells ,Genetic Engineering - Abstract
In this study we determined whether human stromal cells could be engineered with a retroviral vector carrying the interleukin 7 (IL-7) gene and investigated the effects on T cells in vitro and in vivo in a murine model. Transduced mesenchymal cells strongly express CD90 (98.15%), CD105 (87.6%), and STRO-1 (86.7%). IL-7 production was 16.37 (+/-2 SD) pg/ml, which remained stable for 60 days. In vitro-immunoselected naive T cells maintained the CD45RA+ CD45RO- naive phenotype (4.2 times more than controls) after 7 days of culture with IL-7-engineered stromal cells. The apoptosis rate (4.7%) of the naive T cells cultured with transduced stromal cells overlapped with that of freshly isolated cells. Immunohistological analysis detected stromal cells in bone marrow, spleen, and thymus. Cotransplantation of IL-7-engineered stromal cells with CD34+ cells improved engraftment in terms of CD45+ cells and significantly increased the CD3+ cell count in peripheral blood, bone marrow, and spleen. These data demonstrate the following: (1) human stromal cells can be transduced, generating a normal layer; (2) transduced stromal cells in vitro maintain the naive T cell phenotype; and (3) IL-7-transduced stromal cells in vivo home to lymphoid organs and produce sufficient IL-7 in loco, supporting T cell development in a cotransplantation model. Because of their efficient cytokine production and homing, IL-7-engineered stromal cells might be an ideal vehicle to hasten immunological reconstitution in T cell-depleted hosts.
- Published
- 2005
12. Full Haplotype-Mismatched Hematopoietic Stem-Cell Transplantation: A Phase II Study in Patients With Acute Leukemia at High Risk of Relapse
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Adelmo Terenzi, Franca Falzetti, Stelvio Ballanti, Loredana Ruggeri, Flavio Falcinelli, Antonella Santucci, Katia Perruccio, Massimo F. Martelli, Maria Paola Martelli, Cristina Mecucci, Teresa Aloisi, Jean Pierre Saab, Antonio Tabilio, Alessandra Carotti, Rita Felicini, Andrea Velardi, Yair Reisner, and Franco Aversa
- Subjects
Myeloid ,Male ,Oncology ,Cancer Research ,Transplantation Conditioning ,medicine.medical_treatment ,Graft vs Host Disease ,Hematopoietic stem cell transplantation ,Severity of Illness Index ,Recurrence ,Living Donors ,Child ,Acute leukemia ,Leukemia ,Histocompatibility Testing ,Graft Survival ,Hematopoietic Stem Cell Transplantation ,Myeloid leukemia ,Middle Aged ,Fludarabine ,Survival Rate ,Leukemia, Myeloid, Acute ,Treatment Outcome ,surgical procedures, operative ,medicine.anatomical_structure ,Female ,medicine.drug ,Adult ,medicine.medical_specialty ,Adolescent ,ThioTEPA ,Acute ,Risk Assessment ,Internal medicine ,Confidence Intervals ,medicine ,Humans ,Proportional Hazards Models ,business.industry ,Follow-Up Studies ,Multivariate Analysis ,Haplotypes ,medicine.disease ,Transplantation ,Immunology ,business - Abstract
Purpose Establishment of hematopoietic stem-cell (HSC) transplantation from mismatched relatives is feasible for patients with acute leukemia. As our original method of graft processing was unsuitable for large-scale clinical studies, we use automated devices for CD34+ cell purification. Patients and Methods Sixty-seven patients with acute myeloid leukemia (AML; 19 complete remission [CR] 1, 14 CR 2, nine CR > 2, 25 in relapse) and 37 with acute lymphoid leukemia (ALL; 14 CR 1, eight CR 2, two CR > 2, 13 in relapse) were conditioned with total-body irradiation, thiotepa, fludarabine, and antithymocyte globulin. Peripheral-blood progenitor cells were mobilized with recombinant human granulocyte colony-stimulating factor and depleted of T-cells using CD34+ cell immunoselection. No post-transplantation graft-versus-host disease (GvHD) prophylaxis was administered. Results Primary engraftment was achieved in 94 of 101 assessable patients. Six of the seven patients who rejected the primary graft, engrafted after a second transplantation. Overall, 100 of 101 patients engrafted. Acute GvHD developed in eight of 100 patients, and chronic GvHD, in five of 70 assessable patients. Thirty-eight patients died of nonleukemic causes. Relapse occurred in nine of 66 patients receiving transplantation in remission and in 17 of 38 receiving transplantation in relapse. Median follow-up of the 40 patients who survived event-free was 22 months (range, 1 to 65 months). Event-free survival (± standard deviation) rate was 48% ± 8% and 46% ± 10%, respectively, for the 42 AML and 24 ALL patients receiving transplantation in remission. Conclusion Our transplantation procedure provides reliable, reproducible CD34+ cell purification, high engraftment rates, and prevention of GvHD. The mismatched-related transplant emerges as a viable, alternative source of stem cells for acute leukemia patients without matched donors and/or those who urgently need transplantation.
- Published
- 2005
13. Eradication of B-CLL by autologous and allogeneic host nonreactive anti–third-party CTLs
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Rita Krauthgamer, Shraga Aviner, Fabian D. Arditti, Yair Reisner, Arnon Nagler, Alain Berrebi, Benjamin Dekel, Antonio Tabilio, Judith Gan, and Massimo F. Martelli
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Isoantigens ,Fas Ligand Protein ,CD3 Complex ,medicine.medical_treatment ,Chronic lymphocytic leukemia ,Immunology ,Receptors, Antigen, T-Cell ,Apoptosis ,Mice, Transgenic ,Mice, SCID ,Biology ,Biochemistry ,Antibodies ,Mice ,Antigen ,Cancer immunotherapy ,Mice, Inbred NOD ,hemic and lymphatic diseases ,Tumor Cells, Cultured ,medicine ,Animals ,Humans ,Cytotoxic T cell ,fas Receptor ,Lymphocyte function-associated antigen 1 ,Bone Marrow Transplantation ,Mice, Inbred BALB C ,Membrane Glycoproteins ,Cell Biology ,Hematology ,Immunotherapy ,Intercellular Adhesion Molecule-1 ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Lymphocyte Function-Associated Antigen-1 ,Leukemia ,biology.protein ,Antibody ,T-Lymphocytes, Cytotoxic - Abstract
Establishment of cell lines capable of killing leukemia cells, in the absence of alloreactivity against normal host cells, represents a most desirable goal in bone marrow transplantation (BMT) and cancer immunotherapy. By using a human → mouse chimeric model, we demonstrate that allogeneic anti-third-party cytotoxic T lymphocytes (CTLs) depleted of alloreactivity are endowed with a potent anti-B-cell chronic lymphocytic leukemia (B-CLL) reactivity. Likewise, CTL preparations generated from autologous T cells of the same patients with B-CLL exhibited comparable leukemia eradication, suggesting that the reactivity of allogeneic anti-third-party CTLs is not mediated by residual antihost clones. This specificity was also exhibited in vitro, and annexin staining revealed that B-CLL killing is mediated by apoptosis. While the CTLs killing of third-party cells could be blocked by anti-CD3 antibody, the lysis of the B-CLL cells was not inhibited by this antibody, suggesting a T-cell receptor (TCR)-independent cytotoxicity. The role of cell contact leading to apoptosis of B-CLL cells is shown in transwell plates and by anti-lymphocyte function-associated antigen-1 (LFA-1)-blocking antibody. Up-regulation of CD54 and the subsequent apoptosis of B-CLL cells depend on the initial LFA-1/ICAM-1 (intercellular adhesion molecule 1) interaction. Taken together, these results suggest that allogeneic or autologous host nonreactive anti-third-party CTLs may represent a new therapeutic approach for patients with B-CLL. (Blood. 2005;105:3365-3371)
- Published
- 2005
14. Unique human CD133+ leukemia cell line and its modulation towards a mesenchymal phenotype by FGF2 and TGFβ1
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Maria De Ioanni, Lorenzo Moretti, Elisabetta Bonifacio, Antonio Tabilio, Catia Bellucci, Eleonora Lumare, Maria Bodo, Ennio Becchetti, Cinzia Lilli, Tiziano Baroni, Costante Delfini, and Silvia Bellocchio
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FGF2 ,Physiology ,Cellular differentiation ,Osteocalcin ,Clinical Biochemistry ,Bone Marrow Cells ,Core Binding Factor Alpha 1 Subunit ,Receptor, Nerve Growth Factor ,Colony-Forming Units Assay ,Growth factor receptor ,Antigens, CD ,Transforming Growth Factor beta ,Cell Line, Tumor ,Animals ,Humans ,AC133 Antigen ,Collagenases ,RNA, Messenger ,Receptor, Fibroblast Growth Factor, Type 2 ,Autocrine signalling ,Cell Proliferation ,Glycoproteins ,Glycosaminoglycans ,biology ,CD44 ,Mesenchymal stem cell ,leukemia ,TGFbeta ,Cell Differentiation ,Mesenchymal Stem Cells ,Cell Biology ,Precursor Cell Lymphoblastic Leukemia-Lymphoma ,Molecular biology ,Fibronectins ,Haematopoiesis ,biology.protein ,Fibroblast Growth Factor 2 ,Collagen ,Peptides ,A431 cells ,Adult stem cell - Abstract
Immunological features of GM-490 cells, a new blood cell line from a patient with acute lymphoblastic leukemia, included lack of CD34, CD38, CD45, CD14, HLA-DR, and lymphoid and myeloid markers and expression of CD29, CD36, CD44, CD54, CD71, CD105, and CD133. Molecular analysis indicated CD45 gene expression was absent but CD34 mRNA was present. GM-490 cells constitutively produced fibronectin (FN), type III and traces of type I collagen, collagenases, glycosaminoglycans (GAG) and biglycan and betaglycan proteoglycans (PG) as well as FGF2 and TGFβ1. When FGF2 and/or TGFβ1 were added to cells in vitro, they stimulated cell proliferation and differently modulated matrix production and growth factor receptor expression. Reverse transcription-polymerase chain reaction (RT-PCR) detection of transcripts encoding for osteocalcin and RUNX2 suggests GM-490 cells differentiate towards the osteoblast pathway. GM-490 cells expressed the low affinity nerve growth factor receptor (p75LNGFR), a somatic stem cell marker that is not detected in hematopoietic cells, leading to the hypothesis that GM-490 has mesenchymal stem cell properties. The reciprocal modulating effects of FGF2 and TGFβ1 on each other's receptors make the GM-490 cell line a new model for investigating the relationship between these growth factors and their receptors in autocrine loops which are believed to sustain the malignant clone in hematological diseases. © 2005 Wiley-Liss, Inc.
- Published
- 2005
15. Graft engineering for allogeneic haploidentical stem cell transplantation
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Stelvio Ballanti, Franco Aversa, Feliciana Battelli, Roberta Iacucci Ostini, Mauro Di Ianni, Olivia Minelli, Katia Fettucciari, Antonella Santucci, Michele Cimminiello, Antonio Tabilio, Monia Capponi, Tiziana Zei, Massimo F. Martelli, Emanuela Rosati, Carla Silvani, Elisabetta Bonifacio, Maria De Ioanni, Franca Falzetti, and Pierfrancesco Marconi
- Subjects
medicine.medical_treatment ,Transplants ,Lymphoproliferative disorders ,Cell Separation ,Human leukocyte antigen ,Hematopoietic stem cell transplantation ,Lymphocyte Depletion ,Hematopoietic Stem Cell Transplantation ,Humans ,Graft Engineering ,haploidentical Transplantation ,medicine ,Molecular Biology ,Tissue Engineering ,business.industry ,Hematopoietic stem cell ,Cell Biology ,Hematology ,medicine.disease ,Histocompatibility ,Transplantation ,Haematopoiesis ,surgical procedures, operative ,medicine.anatomical_structure ,Immunology ,Molecular Medicine ,Stem cell ,business - Abstract
Haploidentical stem cell transplantation has became a clinical reality in the last 10 years as it provides the chance of transplant for about 50% of patients with hematological malignancies who do not have a matched related or unrelated donor. Proper graft preparation for this type of transplant is crucial and this paper analyses our work over the past decade in the search for the optimal graft processing procedure moving from E-rosetting and soybean agglutination, through a combination of negative or positive selection of hematopoietic stem cells to the current method of one-step positive selection. In preparing a graft for haploidentical transplant, three essential requisites must be met. It must contain (1) a megadose (>10 x 10(6) x kg recipient b.w.) of hematopoietic stem cells to overcome the HLA histocompatibility barrier; (2) very few T-lymphocytes (CD3+ cells < 3 x 10(4)/kg recipient b.w.) to prevent severe acute and chronic graft-versus-host disease (GvHD); (3) very few B-lymphocytes to prevent Epstein-Barr virus-related lymphoproliferative disorders. With current graft processing technologies based on positive selection of hematopoietic stem cells, these requirements can be met. A 70-80% hematopoietic stem cell recovery ensures the target megadose is achieved in over 70% of cases with a T-cell depletion of more than 4 logs and a B-cell depletion of over 3 logs. Progress in graft processing has ensured primary, sustained engraftment rates of over 90% and has significantly reduced the incidence of severe acute GvHD and EBV-related lymphoproliferative disorders. Modern time-saving automated graft processing devices ensure reproducibility, reliability, and biological safety, which make widespread application of the haploidentical transplant currently feasible.
- Published
- 2004
16. Fourth international workshop on haploidentical transplants, Naples, Italy, July 8–10, 2004
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Massimo F. Martelli, Andrea Velardi, Yair Reisner, Franco Locatelli, Stanimir Vuk-Pavlović, Zwi N. Berneman, Antonio Tabilio, and Franco Aversa
- Subjects
medicine.medical_specialty ,Acute leukemia ,Pediatrics ,Bone marrow transplant ,business.industry ,medicine.medical_treatment ,Cell Biology ,Hematology ,Cord Blood Stem Cell Transplantation ,Hematopoietic stem cell transplantation ,medicine.disease ,Surgery ,Transplantation ,Tolerance induction ,surgical procedures, operative ,medicine ,Molecular Medicine ,Human medicine ,business ,Molecular Biology ,DISEASE RELAPSE ,Immunodeficiency - Abstract
Many patients with high-risk hematological malignancies or with incurable inborn errors do not have an HLA-matched sibling and cannot find an HLA-matched donor for an allogeneic hematopoietic stem cell transplantation (SCT). Transplantation strategies using mismatched haploidentical family donors have been an important development. Although the procedure has saved patients from certain death, it is still beset by major problems like life-threatening infections—due to profound immunodeficiency following T-cell depletion and to disease relapse. At every International Workshop on Haploidentical Transplants, new data are presented, showing how scientists are attempting to improve the outcomes of mismatched transplants while reducing the severity of complications. The fourth Workshop continues in this tradition of presenting ground-breaking research. It opened with presentations of the current results with unrelated volunteer and umbilical cord blood transplants and proceeded to a session with the results of haploidentical transplants in the world with series of patients with high-risk acute leukemia, ranging in number from well over 100 in Perugia, Italy, and 80 in Haifa, Israel, to smaller groups in Europe and the United States. The session on graft engineering presented the latest results in the search for the optimal graft. The graft-vs.-leukemia effect in the haploidentical transplant was discussed in depth. Subsequently, attention focussed on one of the major problems in haploidentical transplant, that is, the delay in immunological recovery. The Workshop closed with presentations on tolerance induction that was followed by the results of ongoing registration studies being performed by the Italian GIMEMA group and the European Bone Marrow Transplant group.
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- 2004
17. Endothelial cells in the bone marrow of patients with multiple myeloma
- Author
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Beatrice Nico, Angela Boccarelli, Domenico Ribatti, Francesca Merchionne, Angela Gernone, Mauro Coluccia, Antonio Tabilio, Maria Teresa Petrucci, Claudio Scavelli, Feliciana Battelli, Diego Guidolin, Angelo Vacca, Franco Dammacco, Roberto Ria, and Fabrizio Semeraro
- Subjects
Male ,Umbilical Veins ,Pathology ,medicine.medical_specialty ,Endothelium ,Angiogenesis ,Immunology ,Cell Separation ,Biochemistry ,Neovascularization ,Immunophenotyping ,Vasculogenesis ,Bone Marrow ,Humans ,Medicine ,Aged ,Oligonucleotide Array Sequence Analysis ,Antibacterial agent ,Aged, 80 and over ,Neovascularization, Pathologic ,business.industry ,Gene Expression Profiling ,Cell Biology ,Hematology ,Middle Aged ,Capillaries ,Thalidomide ,Gene Expression Regulation, Neoplastic ,Endothelial stem cell ,Phenotype ,medicine.anatomical_structure ,Case-Control Studies ,Cancer research ,Female ,Endothelium, Vascular ,Bone marrow ,medicine.symptom ,Multiple Myeloma ,business ,Biomarkers - Abstract
Endothelial cells (EC) were extracted through a lectin-based method from bone marrow of 57 patients with active multiple myeloma (MM) and compared with their healthy quiescent counterpart, human umbilical vein EC (HUVEC). MMECs exhibit specific antigens that indicate ongoing angiogenesis and embryo vasculogenesis; solid intercellular connections, hence stability of MM neovessels; and frequent interactions with plasma cells, hence tumor dissemination. They show heterogeneous antigen expression, hence existence of subsets. Their main genetic markers are indicative of a vascular phase. They show intrinsic angiogenic ability, because they rapidly form a capillary network in vitro, and extrinsic ability, because they generate numerous new vessels in vivo. They vividly secrete growth and invasive factors for plasma cells. They signal through kinases mandatory for development of neovascularization. Ultrastructurally, they are abnormal and show metabolic activation, like tumor ECs. Thalidomide heavily interferes with their functions. Vasculogenesis and angiogenesis might contribute to the MM vascular tree and progression, in the form of growth, invasion, and dissemination. In view of the heterogeneity of the antigenic phenotype of MMECs, a mixture (or a sequence) of antiangiogenic agents coupled with thalidomide would seem plausible for the biologic management of MM.
- Published
- 2003
18. Downmodulation of ERK activity inhibits the proliferation and induces the apoptosis of primary acute myelogenous leukemia blasts
- Author
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Antonio Bonati, Antonio Tabilio, Paolo Lunghi, Erminia Ridolo, Carmelo Carlo-Stella, Pierpaolo Dall’Aglio, and Pier Giuseppe Pelicci
- Subjects
Adult ,Male ,MAPK/ERK pathway ,Cancer Research ,Cell cycle checkpoint ,Settore MED/06 - Oncologia Medica ,Down-Regulation ,Apoptosis ,Biology ,Blast Cell Proliferation ,medicine ,Humans ,Enzyme Inhibitors ,Phosphorylation ,Cells, Cultured ,Aged ,Flavonoids ,Mitogen-Activated Protein Kinase 1 ,Mitogen-Activated Protein Kinase Kinases ,Mitogen-Activated Protein Kinase 3 ,Kinase ,G1 Phase ,Cell Differentiation ,Hematology ,Middle Aged ,Flow Cytometry ,Hematopoietic Stem Cells ,medicine.disease ,Leukemia, Myeloid, Acute ,Leukemia ,Oncology ,Caspases ,Mitogen-activated protein kinase ,Cancer research ,biology.protein ,Female ,Mitogen-Activated Protein Kinases ,Poly(ADP-ribose) Polymerases ,Signal transduction ,Cell Division - Abstract
MAP kinase/ERK kinase (MEK)-extracellular signal-regulated kinase (ERK) kinases are frequently activated in acute myelogenous leukemia (AML), and can have prosurvival function. The purpose of this study was to induce downmodulation of MEK-ERK activation in AML primary blasts in order to detect the effect on cell cycle progression and on the apoptosis of leukemic cells. We investigated 14 cases of AML with high ERK 1/2 activity and four cases with undetectable or very low activity. After 24 h incubation of the AML blasts with high ERK activity using PD98059 (New England BioLabs, Beverly, MA, USA), a selective inhibitor of MEK1 phosphorylation, at concentrations of 20 and 40 microM, we observed a strong decrease in the levels of ERK1/2 activity. A significant decrease of blast cell proliferation compared with untreated controls was found. In contrast, the proliferation of blast cells that expressed low or undetectable levels of ERK activity was not inhibited. Time-course analysis demonstrated that the downmodulation of MEK1/2, ERK1 and ERK2 dual-phosphorylation was evident even after 3 h of treatment with 20 and 40 microM. The cleavage of poly(ADP-ribose) polymerase (PARP), an early sign of apoptosis, appeared after 18 h of PD98059 treatment at concentrations of 20 and 40 microM in eight of the 14 cases. After 24 h of treatment, cleaved PARP appeared in all 14 cases. Time-course analysis of cell cycle progression and apoptosis showed that PD98059 induced a G1-phase accumulation with low or undetectable levels of apoptosis after 24 h incubation; after 48 and 72 h incubation, a significant increase of apoptosis was observed. Thus, the primary effect of ERK downmodulation was a cell cycle arrest followed by the apoptosis of a significant percentage of the leukemic blasts. The preclinical model of leukemia treatment reported in this paper makes further comment with regard to MEK1 inhibition as a useful antileukemic target, and encourages the conducting of in vivo studies and clinical investigations.
- Published
- 2003
19. NeoR-Based Transduced T Lymphocytes Detected by Real-Time Quantitative Polymerase Chain Reaction
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Sabrina Di Florio, Antonio Tabilio, G. Venditti, Mauro Di Ianni, Lorenzo Moretti, and Franca Falzetti
- Subjects
Time Factors ,T-Lymphocytes ,Genetic enhancement ,Genetic Vectors ,Immunology ,Biology ,Sensitivity and Specificity ,Thymidine Kinase ,Viral vector ,Transduction (genetics) ,TaqMan ,Humans ,Simplexvirus ,Gene ,Protein Synthesis Inhibitors ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Transfer Techniques ,Reproducibility of Results ,Neomycin ,Genetic Therapy ,U937 Cells ,Hematology ,Suicide gene ,Virology ,Molecular biology ,Kinetics ,Retroviridae ,Real-time polymerase chain reaction ,Drug Resistance, Neoplasm ,Thymidine kinase ,Linear Models - Abstract
To develop a trial with lymphocyte suicide gene therapy in patients with hematological malignancies, we transduced human T lymphocytes with a retroviral vector (LSN-tk) encoding the herpes simplex virus thymidine kinase (tk) and the neomycin resistance (NeoR) genes. Precise quantification of gene transfer is crucial for any gene therapy protocol, but methods using semiquantitative PCR are inaccurate and subject to variations. Real-time quantitative PCR could be a valid alternative. A TaqMan probe was designed to hybridize with the NeoR gene. The PCR product is 64 nucleotides long and readily quantified by TaqMan probe binding. The analysis was performed soon after transduction and repeated after the selection procedure. This method was more accurate, reproducible, and sensitive than the semiquantitative PCR assay. Accuracy was the same whether the analysis was performed at the highest rate or at the lowest rate of transduction. Additionally we used real-time PCR to monitor the kinetics of enrichment of the transduced cells over the selection time and showed how 7 days of selection are needed. This study precisely quantified the percentages of cells transduced by the retroviral vector and could have major implications in gene therapy studies.
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- 2003
20. Megadose of hematopoietic stem cells for haploidentical transplants
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Loredana Ruggeri, Andrea Velardi, Yair Reisner, Esther Bachar-Lustig, Antonio Tabilio, Shlomit Reich-Zeliger, Hilit Gur, Massimo F. Martelli, and Franco Aversa
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Transplantation ,Haematopoiesis ,surgical procedures, operative ,business.industry ,Immunology ,Immunology and Allergy ,Medicine ,Human leukocyte antigen ,Progenitor cell ,Stem cell ,business - Abstract
Studies in mice and humans demonstrate that transplantation of hematopoietic progenitors in numbers larger than commonly used (“megadose” transplants) overcomes major genetic barriers without induction of graft-versus-host disease. Therefore, transplantation from a human leukocyte antigen mismatched
- Published
- 2002
21. Treatment of adult acute lymphoblastic leukemia (ALL): long-term follow-up of the GIMEMA ALL 0288 randomized study
- Author
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Giorgina Specchia, Felicetto Ferrara, Luciana Annino, Stefania Ciolli, Antonio Peta, Antonio Tabilio, Simona Sica, Francesco Di Raimondo, Andrea Camera, Giuseppe Visani, Rosangela Invernizzi, Giuseppe Fioritoni, Wilma Deplano, Nicola Cascavilla, Franco Mandelli, Francesco Fabbiano, Pietro Leoni, Bruno Rotoli, Michele Baccarani, M. L. Vegna, and Sergio Amadori
- Subjects
Adult ,Male ,medicine.medical_specialty ,Myeloid ,Adolescent ,Cyclophosphamide ,medicine.medical_treatment ,Immunology ,Biochemistry ,Gastroenterology ,Disease-Free Survival ,chemistry.chemical_compound ,Prednisone ,Acute lymphocytic leukemia ,Internal medicine ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Humans ,Longitudinal Studies ,Child ,Survival analysis ,Aged ,Chemotherapy ,business.industry ,Remission Induction ,Cell Biology ,Hematology ,Middle Aged ,Precursor Cell Lymphoblastic Leukemia-Lymphoma ,Prognosis ,medicine.disease ,Survival Analysis ,Nitrogen mustard ,Surgery ,medicine.anatomical_structure ,chemistry ,Adult Acute Lymphoblastic Leukemia ,Female ,business ,medicine.drug - Abstract
The GIMEMA ALL 0288 trial was designed to evaluate the impact of a 7-day prednisone (PDN) pretreatment on complete remission (CR) achievement and length, the influence of the addition of cyclophosphamide (random I) to a conventional 4-drug induction on CR rate and duration, and whether an early post-CR intensification (random II) by an 8-drug consolidation could improve CR duration. Median follow-up of this study was 7.3 years. From January 1988 to April 1994, among 794 adult (> 12 but
- Published
- 2002
22. ALLOGENEIC TRANSPLANTATION ACROSS THE HLA BARRIERS
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Andrea Velardi, Massimo F. Martelli, Antonio Tabilio, and Franco Aversa
- Subjects
Allogeneic transplantation ,Pilot Projects ,Disease ,Human leukocyte antigen ,barriere hla ,trapianto allogenico ,medicine ,Humans ,Transplantation, Homologous ,Acute leukemia ,Leukemia ,Haploidentical transplantation ,business.industry ,Histocompatibility Testing ,Hematopoietic Stem Cell Transplantation ,Hematopoietic stem cell ,Hematology ,Haematopoiesis ,surgical procedures, operative ,medicine.anatomical_structure ,Haplotypes ,Acute Disease ,Immunology ,Stem cell ,business - Abstract
In high-risk acute leukemia patients, a 10-fold increase in the dose of extensively T-cell-depleted hematopoietic stem cells ensures sustained full-donor engraftment of one-haplotype-mismatched transplants without graft-vs.-host disease. Since our first successful pilot study, which exploited the principle of a megadose stem cell transplant, our efforts have concentrated on developing new conditioning regimens, optimizing graft processing and improving the post-transplant immunologic recovery. The results so far achieved in more than 100 high-risk acute leukemia patients show that haploidentical transplantation is now a clinical reality. Because virtually all patients in need of a hematopoietic stem cell transplant have a full-haplotype-mismatched family donor, a T-cell-depleted mismatched transplant can be offered with curative intent, thus extending allogeneic transplantation procedures to virtually all candidates.
- Published
- 2001
23. Results of a Prospective Study with High-dose Etoposide, Thiotepa and Carboplatin and Peripheral Blood Stem Cell Rescue for High-risk Stage II-IIIA and Selected Stage IV Breast Cancer Patients
- Author
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C. Basurto, Cynthia Aristei, Maurizio Tonato, Paolo Latini, M. Colozza, Roberta Cherubini, Antonio Tabilio, Franca Falzetti, Anna Maria Mosconi, Stefania Gori, and Massimo F. Martelli
- Subjects
Adult ,Risk ,Oncology ,Cancer Research ,medicine.medical_specialty ,medicine.medical_treatment ,Breast Neoplasms ,ThioTEPA ,Drug Administration Schedule ,Carboplatin ,030218 nuclear medicine & medical imaging ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Breast cancer ,Internal medicine ,Antineoplastic Combined Chemotherapy Protocols ,Granulocyte Colony-Stimulating Factor ,medicine ,Humans ,cancer ,Prospective Studies ,breast ,Etoposide ,Neoplasm Staging ,Chemotherapy ,business.industry ,Induction chemotherapy ,General Medicine ,Middle Aged ,medicine.disease ,Chemotherapy regimen ,Regimen ,Treatment Outcome ,chemistry ,030220 oncology & carcinogenesis ,Female ,business ,Thiotepa ,medicine.drug - Abstract
Aims and Background To investigate the safety and efficacy of a high-dose chemotherapy regimen with etoposide, carboplatin and thiotepa in high-risk stage II-IIIA breast cancer and in responsive metastatic patients. Study Design From April 1992 to December 1998, 24 patients with high-risk stage II-IIIA breast cancer (≥9 positive nodes) and 9 responsive metastatic patients were enrolled in the trial. After induction chemotherapy with an anthracycline-based regimen, peripheral blood stem cells were mobilized with cyclophosphamide (7 g/m2) and G-CSF (5-16 μg/kg/sc/day). The high-dose chemotherapy regimen consisted of etoposide (1000 mg/m2), carboplatin (800 mg/m2) and thiotepa (500 mg/m2). At the end of the high-dose chemotherapy, all stage II-IIIA patients received radiotherapy to the breast or chest wall and draining nodes; stage IV patients were irradiated to sites of disease, if feasible. All ER+ and/or PgR+ patients were treated with hormone therapy. Results For stage II-IIIA high-risk patients, the median follow-up was 4.36 years (range, 1.93-6.94), and the Kaplan-Meier estimate at 5 years of disease-free survival and overall survival was 54.8 ± 11% SE and 76.73 ± 9.4% SE, respectively. For metastatic patients, the median follow-up was 4.93 years (range, 4.15-7.95), and the Kaplan-Meier estimate at 5 years of progression-free survival and overall survival was 22.2 ± 13.9% SE and 76.2 ± 14.8% SE, respectively. No treatment-related deaths were observed. Conclusions Our results are comparable to those obtained in other high-dose chemotherapy trials but do not seem to be superior to conventional-dose therapy given to similar patients.
- Published
- 2001
24. Postgrafting administration of granulocyte colony-stimulating factor impairs functional immune recovery in recipients of human leukocyte antigen haplotype–mismatched hematopoietic transplants
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Franco Aversa, Katia Perruccio, Massimo F. Martelli, Isabella Volpi, Antonella Tosti, Antonio Tabilio, Marusca Capanni, Luigina Romani, Loredana Ruggeri, Sabrina Posati, and Andrea Velardi
- Subjects
Adult ,Male ,Adolescent ,Immunology ,Antigen presentation ,Graft vs Host Disease ,Human leukocyte antigen ,Biology ,medicine.disease_cause ,Biochemistry ,Th2 Cells ,Immune system ,Immunopathology ,Granulocyte Colony-Stimulating Factor ,medicine ,Humans ,Child ,Candida ,Salvage Therapy ,Leukemia ,Graft Survival ,Hematopoietic Stem Cell Transplantation ,Immunologic Deficiency Syndromes ,Receptors, Interleukin-12 ,Dendritic Cells ,Receptors, Interleukin ,Cell Biology ,Hematology ,Middle Aged ,Immune dysregulation ,Interleukin-12 ,Hematopoietic Stem Cell Mobilization ,CD4 Lymphocyte Count ,Interleukin-10 ,Granulocyte colony-stimulating factor ,Transplantation ,Protein Subunits ,Aspergillus ,Treatment Outcome ,Haplotypes ,Child, Preschool ,Acute Disease ,Interleukin 12 ,Female ,Disease Susceptibility ,Interleukin-4 ,Immunologic Memory - Abstract
In human leukocyte antigen haplotype–mismatched transplantation, extensive T-cell depletion prevents graft-versus-host disease (GVHD) but delays immune recovery. Granulocyte colony-stimulating factor (G-CSF) is given to donors to mobilize stem cells and to recipients to ensure engraftment. Studies have shown that G-CSF promotes T-helper (Th)-2 immune deviation which, unlike Th1 responses, does not protect against intracellular pathogens and fungi. The effect of administration of G-CSF to recipients of mismatched hematopoietic transplants with respect to transplantation outcome and functional immune recovery was investigated. In 43 patients with acute leukemia who received G-CSF after transplantation, the engraftment rate was 95%. However, the patients had a long-lasting type 2 immune reactivity, ie, Th2-inducing dendritic cells not producing interleukin 12 (IL-12) and high frequencies of IL-4– and IL-10–producing CD4+ cells not expressing the IL-12 receptor β2 chain. Similar immune reactivity patterns were observed on exposure of donor cells to G-CSF. Elimination of postgrafting administration of G-CSF in a subsequent series of 36 patients with acute leukemia, while not adversely affecting engraftment rate (93%), resulted in the anticipated appearance of IL-12–producing dendritic cells (1-3 months after transplantation versus > 12 months in transplant recipients given G-CSF), of CD4+ cells of a mixed Th0/Th1 phenotype, and of antifungal T-cell reactivity in vitro. Moreover, CD4+ cell counts increased in significantly less time. Finally, elimination of G-CSF–mediated immune suppression did not significantly increase the incidence of GVHD (< 15%). Thus, this study found that administration of G-CSF to recipients of T-cell–depleted hematopoietic transplants was associated with abnormal antigen-presenting cell functions and T-cell reactivity. Elimination of postgrafting administration of G-CSF prevented immune dysregulation and accelerated functional immune recovery.
- Published
- 2001
25. Polymeric protein-polyamine conjugates: A new class of uremic toxins affecting erythropoiesis
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Alessandro Lentini, Simone Beninati, Antonio Tabilio, Francesco Galli, Franco Canestrari, Umberto Buoncristiani, and Serena Benedetti
- Subjects
erythroid cells ,Macromolecular Substances ,Spermidine ,Anemia ,Tissue transglutaminase ,Size-exclusion chromatography ,In Vitro Techniques ,Pharmacology ,Colony-Forming Units Assay ,chemistry.chemical_compound ,Renal Dialysis ,mental disorders ,Polyamines ,medicine ,Humans ,Erythropoiesis ,CFU-E ,Aged ,Toxins, Biological ,Uremia ,chemistry.chemical_classification ,hemodialysis ,end-stage renal disease ,biology ,Blood Proteins ,Middle Aged ,Hematopoietic Stem Cells ,medicine.disease ,anemia ,In vitro ,Enzyme ,chemistry ,Biochemistry ,Nephrology ,biology.protein ,Kidney Failure, Chronic ,Polyamine - Abstract
Polymeric protein-polyamine conjugates: A new class of uremic toxins affecting erythropoiesis. Background Preliminary evidence on the accumulation of polyamine-protein conjugates (PPCs) was obtained in uremic patients. The presence of these substances in the plasma of hemodialysis (HD) patients was evaluated, and their possible contribution to uremic anemia was investigated by testing the effect of PPC synthesized in vitro on erythroid cell proliferation. Methods Plasma PPC was measured by high-performance liquid chromatography. The in vitro synthesis of PPC from human plasma was carried out by means of the enzyme transglutaminase in the presence of either [3H]-labeled or unlabeled spermidine (SPD). After gel filtration chromatography and detection of the fractions containing [3H]SPD. the latter were tested for their effect on monouclear bone marrow cell proliferation. Results In three out of four patients examined, mainly SPD-protein conjugates (SPD-PC) were observed to accumulate during HD. The levels ranged from 0.17 to 4.93 pmol/mg proteins before dialysis, and these values increased at 30 minutes and at the end of the dialysis up to levels 11.90 pmol/mg. SPD-PC levels in healthy controls were 1.46 ± 0.82. SPD-PCs synthesized in vitro were recovered in two main fractions showing a molecular weight of > 100KD (peak 1) and of approximately 30 to 50 KD (peak 3), respectively. The SPD-PC contained in peak 1 showed the greatest inhibitory effect on colony-forming units-erythroid (CFU-E) proliferation without any appreciable effect on burst-forming units-erythroid (BFU-E). Conclusion We demonstrate that SPD-PC can accumulate in HD patients. These substances, which affect CFU-E proliferation, can be considered as an at yet unrevealed class of uremictoxins contributing to the onset of the uremic anemia.
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- 2001
26. Limited engraftment capacity of bone marrow–derived mesenchymal cells following T-cell–depleted hematopoietic stem cell transplantation
- Author
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Vittorio Rizzoli, Massimo F. Martelli, E. Regazzi, Gabriella Sammarelli, Antonio Tabilio, Daniela Cilloni, Carmelo Carlo-Stella, Franca Falzetti, Franco Aversa, and Simona Colla
- Subjects
Pathology ,medicine.medical_specialty ,Stromal cell ,T cell ,medicine.medical_treatment ,Immunology ,Mesenchymal stem cell ,Cell Biology ,Hematology ,Human leukocyte antigen ,Hematopoietic stem cell transplantation ,Biology ,Biochemistry ,Molecular biology ,medicine.anatomical_structure ,medicine ,Bone marrow ,Stem cell ,Progenitor cell - Abstract
The engraftment capacity of bone marrow–derived mesenchymal cells was investigated in 41 patients who had received a sex-mismatched, T-cell–depleted allograft from human leukocyte antigen (HLA)–matched or –mismatched family donors. Polymerase chain reaction (PCR) analysis of the human androgen receptor (HUMARA) or the amelogenin genes was used to detect donor-derived mesenchymal cells. Only 14 marrow samples (34%) from 41 consenting patients generated a marrow stromal layer adequate for PCR analysis. Monocyte-macrophage contamination of marrow stromal layers was reduced below the levels of sensitivity of HUMARA and amelogenin assays (5% and 3%, respectively) by repeated trypsinizations and treatment with the leucyl-leucine (leu-leu) methyl ester. Patients who received allografts from 12 female donors were analyzed by means of the HUMARA assay, and in 5 of 12 cases a partial female origin of stromal cells was demonstrated. Two patients who received allografts from male donors were analyzed by amplifying the amelogenin gene, and in both cases a partial male origin of stromal cells was shown. Fluorescent in situ hybridization analysis using a Y probe confirmed the results of PCR analysis and demonstrated in 2 cases the existence of a mixed chimerism at the stromal cell level. There was no statistical difference detected between the dose of fibroblast progenitors (colony-forming unit–F [CFU-F]) infused to patients with donor- or host-derived stromal cells (1.18 ± 0.13 × 104/kg vs 1.19 ± 0.19 × 104/kg; P ≥ .97). In conclusion, marrow stromal progenitors reinfused in patients receiving a T-cell–depleted allograft have a limited capacity of reconstituting marrow mesenchymal cells.
- Published
- 2000
27. Limited engraftment capacity of bone marrow–derived mesenchymal cells following T-cell–depleted hematopoietic stem cell transplantation
- Author
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Daniela Cilloni, Carmelo Carlo-Stella, Franca Falzetti, Gabriella Sammarelli, Ester Regazzi, Simona Colla, Vittorio Rizzoli, Franco Aversa, Massimo F. Martelli, and Antonio Tabilio
- Subjects
Immunology ,Cell Biology ,Hematology ,Biochemistry - Abstract
The engraftment capacity of bone marrow–derived mesenchymal cells was investigated in 41 patients who had received a sex-mismatched, T-cell–depleted allograft from human leukocyte antigen (HLA)–matched or –mismatched family donors. Polymerase chain reaction (PCR) analysis of the human androgen receptor (HUMARA) or the amelogenin genes was used to detect donor-derived mesenchymal cells. Only 14 marrow samples (34%) from 41 consenting patients generated a marrow stromal layer adequate for PCR analysis. Monocyte-macrophage contamination of marrow stromal layers was reduced below the levels of sensitivity of HUMARA and amelogenin assays (5% and 3%, respectively) by repeated trypsinizations and treatment with the leucyl-leucine (leu-leu) methyl ester. Patients who received allografts from 12 female donors were analyzed by means of the HUMARA assay, and in 5 of 12 cases a partial female origin of stromal cells was demonstrated. Two patients who received allografts from male donors were analyzed by amplifying the amelogenin gene, and in both cases a partial male origin of stromal cells was shown. Fluorescent in situ hybridization analysis using a Y probe confirmed the results of PCR analysis and demonstrated in 2 cases the existence of a mixed chimerism at the stromal cell level. There was no statistical difference detected between the dose of fibroblast progenitors (colony-forming unit–F [CFU-F]) infused to patients with donor- or host-derived stromal cells (1.18 ± 0.13 × 104/kg vs 1.19 ± 0.19 × 104/kg; P ≥ .97). In conclusion, marrow stromal progenitors reinfused in patients receiving a T-cell–depleted allograft have a limited capacity of reconstituting marrow mesenchymal cells.
- Published
- 2000
28. Retrovirus-mediated transfer of the herpes simplex virus thymidine kinase and enhanced green fluorescence protein genes in primary T lymphocytes
- Author
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Mauro Di Ianni, Sabrina Di Florio, Pier Giuseppe Pelicci, Carla Sebastiani, G. Venditti, Antonio Tabilio, Diego Alfonsi, and Marta Fagioli
- Subjects
Interleukin 2 ,Time Factors ,T-Lymphocytes ,Genetic Vectors ,Green Fluorescent Proteins ,Biology ,Transfection ,medicine.disease_cause ,Antiviral Agents ,Thymidine Kinase ,Virus ,Viral vector ,medicine ,Humans ,Simplexvirus ,Ganciclovir ,Genetic transfer ,Hematology ,T lymphocyte ,Suicide gene ,Flow Cytometry ,Virology ,Luminescent Proteins ,Herpes simplex virus ,Thymidine kinase ,Interleukin-2 ,Cell Division ,medicine.drug - Abstract
The EGFP-tk retroviral vector, encoding enhanced green fluorescent protein (EGFP) and the herpes simplex virus thymidine kinase (HSV-tk) packaged in a Phoenix amphotropic cell line, was used to transduce healthy donor T lymphocytes. Infection yielded a mean of 41.8 +/- 9.3% SD (range 31.1-48.4%) EGFP-positive cells and a mean of 92 +/- 2% SD (range 90-94%) after cell sorting. EGFP expression remained stable for 30 d after infection. The entire gene transfer procedure had no significant effect on lymphocyte subsets and slightly reduced clonogenicity. Ganciclovir (gcv) treatment (1 microg/ml x 10 d) killed all EGFP-positive cells in the transduced and transduced/sorted populations, but had no effect on untransduced controls. Our results show that primary T lymphocytes can be transduced using an EGFP-tk vector that yields a homogeneous infected population without affecting lymphocyte subsets, function and clonogenicity.
- Published
- 2000
29. A New Methodology of Fetal Stem Cell Isolation, Purification, and Expansion: Preliminary Results for Noninvasive Prenatal Diagnosis
- Author
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Francesca Tilesi, Antonio Tabilio, Giuliana Coata, Gian Carlo Di Renzo, Vincenzo Lauro, and Luana Pennacchi
- Subjects
Male ,Pathology ,medicine.medical_specialty ,Immunology ,Cell Culture Techniques ,CD34 ,Antigens, CD34 ,Biology ,Peripheral blood mononuclear cell ,Stem Cell Isolation ,Pregnancy ,Prenatal Diagnosis ,medicine ,Humans ,Progenitor cell ,In Situ Hybridization, Fluorescence ,Erythroid Precursor Cells ,Cloning ,Fetus ,Immunomagnetic Separation ,Stem Cells ,Hematology ,Fetal Blood ,Molecular biology ,In vitro ,Karyotyping ,Pregnancy Trimester, Second ,Female ,DNA Probes ,Chromosome 21 ,Cell Division - Abstract
We developed a combined methodological approach to enrich and to proliferate in vitro fetal CD34+ stem progenitor cells. Using a magnetic cell-sorting technique, CD34+ cells from pregnant women at the early-second trimester were isolated and enriched and compared to those isolated from blood of nonpregnant women. The number and frequency of CD34+ cells were significantly higher (p0.001) in the pregnant women. Unenriched peripheral blood mononuclear cells (PBMC) and enriched CD34+ cells were cultured in a methylcellulose system to evaluate the cloning potential of progenitor cells. After culture, the numbers of burst-forming units erythroid/colony-forming units erythroid (BFU-E/CFU-E) and colony-forming units granulocyte-macrophage (CFU-GM) colonies were increased by 33 and 16 times, respectively. Finally, to distinguish between fetal and maternal cells, four cases of cultured cells were hybridized with specific probes for X and Y chromosomes and two cases with a specific probe for chromosome 21. In normal pregnancies, we identified a high number of male fetal cells and an elevated fetal/maternal ratio. When we analyzed blood samples from pregnancies with trisomic fetuses, we scored a high ratio of trisomic cells respect to maternal cells that was significantly different from the ratio of pregnancies with normal fetuses. Our results demonstrate fetal progenitor cells may be cultured and detected successfully with an appropriate combined methodological approach, which may significantly increase the feasibility of noninvasive prenatal diagnosis.
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- 2000
30. Peripheral blood progenitor cell mobilization in healthy donors receiving recombinant human granulocyte colony-stimulating factor
- Author
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Carmelo Carlo-Stella, F. Falzetti, Antonio Tabilio, Vittorio Rizzoli, Massimo F. Martelli, Clara Cesana, E. Regazzi, and Franco Aversa
- Subjects
Adult ,Male ,Cancer Research ,CFU-GM ,CD34 ,Blood Donors ,Andrology ,Granulocyte Colony-Stimulating Factor ,Genetics ,medicine ,Humans ,Transplantation, Homologous ,Progenitor cell ,Molecular Biology ,business.industry ,Hematopoietic Stem Cell Transplantation ,Cell Biology ,Hematology ,Leukapheresis ,Middle Aged ,Hematopoietic Stem Cell Mobilization ,Recombinant Proteins ,Granulocyte colony-stimulating factor ,Haematopoiesis ,medicine.anatomical_structure ,Immunology ,Female ,Bone marrow ,Stem cell ,business - Abstract
Objective We analyzed the incidence of primitive (LTC-IC) and committed (CFU-mix, BFU-E, CFU-GM) hematopoietic progenitors detected under steady-state conditions and upon progenitor cell mobilization in a cohort of healthy donors receiving recombinant human granulocyte colony-stimulating factor (rhG-CSF). Materials and methods Healthy donors ( n = 30) of HLA-mismatched or -matched stem cell transplants were mobilized with rhG-CSF (8 μg/Kg body weight subcutaneously twice daily until completion of leukapheresis). PBPC collections were started after 4 days of rhG-CSF therapy. Results Steady-state incidence of bone marrow LTC-IC, but not committed progenitors, significantly correlated with the numbers of mobilized CD34 + cells ( r = 0.6, p = 0.004), CFU-GM ( r = 0.79, p = 0.0005) and CFC ( r = 0.76, p = 0.001) detected after 4 days of rhG-CSF therapy. Statistically significant correlations were also found between steady-state blood CFU-GM and peak numbers of CD341 cells ( r = 0.68, p = 0.001), numbers of day 4 CD341 cells ( r = 0.52, p = 0.005), CFU-GM ( r = 0.63, p = 0.002), and CFC ( r = 0.61, p = 0.003). Conclusion Our data show that in normal volunteers baseline marrow LTC-IC and blood CFU-GM correlate with rhG-CSF-mobilized PBPC. The potential clinical relevance of these findings in the identification of poor mobilizers will be tested in a prospective study.
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- 2000
31. T Lymphocyte Transduction with Herpes Simplex VirusThymidine Kinase (HSV-tk) Gene: Comparison of Four Different Infection Protocols
- Author
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Massimo F. Martelli, P. Mannoni, Sabrina Di Florio, Franca Falzetti, G. Venditti, Mauro Di Ianni, and Antonio Tabilio
- Subjects
Interleukin 2 ,Cell Survival ,T-Lymphocytes ,Genetic Vectors ,Immunology ,Centrifugation ,Biology ,Lymphocyte Activation ,Transfection ,medicine.disease_cause ,Polymerase Chain Reaction ,Thymidine Kinase ,Immunophenotyping ,Viral vector ,Viral Proteins ,Transduction (genetics) ,HSV-Tk Gene ,Genes, Reporter ,medicine ,Simplexvirus ,Phytohemagglutinins ,Ganciclovir ,Cells, Cultured ,Kanamycin Kinase ,Reproducibility of Results ,Drug Resistance, Microbial ,Hematology ,T lymphocyte ,Virology ,Molecular biology ,Coculture Techniques ,Fibronectins ,Herpes simplex virus ,Thymidine kinase ,Culture Media, Conditioned ,Enzyme Induction ,Interleukin-2 ,Gentamicins ,Moloney murine leukemia virus ,Safety ,Cell Division ,medicine.drug - Abstract
In this study, we assessed the efficiency of T lymphocyte transduction with a retroviral vector carrying the herpes simplex virus thymidine kinase (HSV-tk) and neomycin phosphotransferase (neo) genes by four different protocols: standard supernatant infection, supernatant infection plus centrifugation steps, supernatant infection on fibronectin fragment-coated wells, and cocultivation. After retrovirus-mediated gene transfer of tk-neo in PHA/IL-2-stimulated primary T lymphocytes and G418 selection, T cells retained their proliferative activity, alloresponsiveness, ability to produce and to respond to IL-2, and ganciclovir (gcv)-specific sensitivity. When the four different transduction techniques were compared, no significant differences were seen in terms of cellular viability, proliferation capacity, and immunophenotyping. tk gene expression was the same in all transduced selected populations, as indicated by gcv sensitivity. Transduction efficiency was evaluated by semiquantitative PCR. Using the standard supernatant infection method, the rate of infection was extremely low (
- Published
- 1999
32. Phase III efficacy study of interleukin-3 after autologous bone marrow transplantation in patients with malignant lymphoma
- Author
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Dominique Maraninchi, Willem E. Fibbe, Beate Kiese, Antonio Tabilio, I. Sklenar, Richard Herrmann, Simon Durrant, Edo Vellenga, Koos H. Zwinderman, José‐Luis Pico Volonte, Rolf E. Brouwer, W. R. Bezwoda, Donald Milligan, and D. A. Winfield
- Subjects
Chemotherapy ,medicine.medical_specialty ,Neutrophil Engraftment ,Platelet Engraftment ,business.industry ,medicine.medical_treatment ,Hematology ,medicine.disease ,Placebo ,Gastroenterology ,Lymphoma ,Surgery ,Discontinuation ,medicine.anatomical_structure ,Internal medicine ,Medicine ,Bone marrow ,business ,Adverse effect - Abstract
We evaluated the efficacy of recombinant human interleukin-3 (rhIL-3) in reducing the number of platelet transfusions and major infections after autologous bone marrow transplantation (ABMT) in patients with malignant lymphoma. 198 patients with non-Hodgkin's lymphoma (NHL, n = 111) and Hodgkin's disease (HD, n = 87) were randomized to receive rhIL-3 10 μg/kg/d (n = 130) or placebo (n = 68) for a maximum of 28 d after ABMT. Several well-known conditioning regimens were used. From day 1 after ABMT patients were treated with placebo or rhIL-3 at a dose of 10 μg/kg/d by continuous i.v. infusion for 7 d and then by s.c. administration for 21 d or until platelet (50 × 109/l) and neutrophil (0.5 × 109/l) recovery had occurred. Treatment was completed in 54% of the patients in the rhIL-3 group versus 75% in the placebo group (P
- Published
- 1999
33. Effects of the Tyrosine Kinase Inhibitor AG957 and an Anti-Fas Receptor Antibody on CD34+ Chronic Myelogenous Leukemia Progenitor Cells
- Author
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Alexander Levitzki, Gabriella Sammarelli, Barbara Savoldo, Carmelo Carlo-Stella, Vittorio Rizzoli, Simona Colla, Daniela Cilloni, D. Garau, Aviv Gazit, Antonio Tabilio, and E. Regazzi
- Subjects
ABL ,medicine.drug_class ,Immunology ,breakpoint cluster region ,Tyrosine phosphorylation ,Cell Biology ,Hematology ,Biology ,medicine.disease ,Biochemistry ,Tyrosine-kinase inhibitor ,chemistry.chemical_compound ,chemistry ,Cell surface receptor ,hemic and lymphatic diseases ,medicine ,Cancer research ,Growth inhibition ,Tyrosine kinase ,Chronic myelogenous leukemia - Abstract
The hallmark of chronic myelogenous leukemia (CML) is the Philadelphia (Ph) chromosome that fuses genetic sequences of the BCR gene on chromosome 22 with c-ABL sequences translocated from chromosome 9. BCR/ABL fusion proteins have a dysregulated protein tyrosine kinase (PTK) activity exerting a key role in malignant transformation. Targeting the tyrosine kinase activity of BCR/ABL or using agents capable of triggering apoptosis might represent attractive therapeutic approaches for ex vivo purging. AG957, a member of the tyrphostin compounds, exerts a selective inhibition of p210BCR/ABLtyrosine phosphorylation. We report here that preincubation of CML or normal CD34+ cells with graded concentration of AG957 (1 to 100 μmol/L) resulted in a statistically significant, dose-dependent suppression of colony growth from multipotent, erythroid, and granulocyte-macrophage progenitors as well as the more primitive long-term culture-initiating cells (LTC-IC). However, AG957 doses causing 50% inhibition (ID50) of CML and normal progenitors were significantly different for multilineage colony-forming units (CFU-Mix; 12 v 64 μmol/L; P = .008), burst-forming unit-erythroid (BFU-E; 29 v 89 μmol/L;P = .004), colony-forming unit–granulocyte-macrophage (CFU-GM; 34 v 85 μmol/L; P = .004), and LTC-IC (43 v 181 μmol/L; P = .004). In 5 of 10 patients, analysis of BCR/ABL mRNA on single progenitors by reverse transcription-polymerase chain reaction showed that AG957 at 50 μmol/L significantly reduced the mean (±SD) percentage of BCR/ABL-positive progenitors (92% ± 10% v 33 ± 5%;P = .001). Because AG957 treatment resulted in significantly higher percentages of apoptotic cells (30% v9%) in the BCR/ABL-transfected 32DLG7 cells as compared with 32D-T2/93 cells (BCR/ABL-negative), we investigated the combined effects of AG957 with the anti-Fas receptor (Fas-R) monoclonal antibody CH11 that triggers apoptosis. As compared with AG957 alone, the sequential treatment of CML CD34+ cells with AG957 (1 μmol/L) and CH11 (1 μg/mL) increased CFU-Mix, BFU-E, and CFU-GM growth inhibition by 1.6-fold, 3-fold, and 4-fold, respectively. In contrast, the treatment of normal CD34+ cells with AG957 and CH11 failed to enhance AG957-induced colony growth inhibition. We conclude that (1) AG957 inhibits in a dose-dependent manner CML CD34-derived colony formation by both primitive LTC-IC as well as committed CFU-Mix, BFU-E, and CFU-GM; (2) this growth inhibition is associated with the selection of a substantial amount of BCR/ABL-negative progenitors; and (3) the antiproliferative effect of AG957 is dramatically increased by combining this compound with the anti–Fas-R antibody CH11. These data may have significant therapeutic applications.
- Published
- 1999
34. Molecular Biology of Hematopoiesis 6
- Author
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Nader G. Abraham, Antonio Tabilio, Massimo Martelli, Shigetaka Asano, Alberto Donfrancesco, Nader G. Abraham, Antonio Tabilio, Massimo Martelli, Shigetaka Asano, and Alberto Donfrancesco
- Subjects
- Oncology, Endocrinology, Neurology, Cancer, Medical genetics, Anatomy
- Abstract
This volume of Molecular Biology of Hematopoiesis is dedicated to many inter national scientists and clinicians for their contribution to the field of Hematology/ Oncology presented at the 11th International Symposium on Molecular Biology of Hematopoiesis, which was held in Bormio, Italy, June 25-29, 1998. The continuous support of the Presidents of the meeting, Professor F. Takaku, President of Jichi University, and E. D. Thomas, Nobel Laureate, was greatly acknowledged, especially Professor Takaku, for his vision and support for development of gene therapy in Japan. New information on BMT for autoimmune disease and organ transplantation was presented at the symposium and is published in this volume. Several new findings on gene therapy/transfer into HSC were presented by E. F. Vanin and A. Nienhuis, K. Humphries, 1. A. Nolta, H. E. Heslop, and M. K. Brenner. Professors S. Asano and K. Tani presented new studies on gene transfer into primates. Among the highlights were the new papers on gene transfer presented by G. Wage maker, N. G. Abraham, and M. Onoderea from R. M. BJaese's group. The use of BMT for organ transplant and autoim mune disease was updated and a representative paper is presented in this volume.
- Published
- 2012
35. Reverse transcription polymerase chain reaction is a reliable assay for detecting leukemic colonies generated by chronic myelogenous leukemia cells
- Author
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Carmelo Carlo-Stella, Gabriella Sammarelli, G. P. Dotti, Barbara Savoldo, D. Cilloni, Antonio Tabilio, E. Regazzi, D. Garau, and Vittorio Rizzoli
- Subjects
Adult ,Male ,Cancer Research ,Transcription, Genetic ,Karyometry ,Fusion Proteins, bcr-abl ,Chromosomal translocation ,Biology ,Philadelphia chromosome ,Polymerase Chain Reaction ,law.invention ,Colony-Forming Units Assay ,law ,Leukemia, Myelogenous, Chronic, BCR-ABL Positive ,medicine ,Humans ,Polymerase chain reaction ,Aged ,DNA Primers ,Hematology ,Middle Aged ,Hematopoietic Stem Cells ,medicine.disease ,Molecular biology ,Reverse transcriptase ,Chromosome Banding ,Reverse transcription polymerase chain reaction ,Real-time polymerase chain reaction ,Oncology ,Female ,Stem cell ,Chronic myelogenous leukemia - Abstract
Single-colony karyotyping (SCK) and reverse-transcription polymerase chain reaction (RT-PCR) are two increasingly used techniques for the quantification of leukemic colonies generated by chronic myelogenous leukemia (CML) cell fractions purged or selected in vitro. Recently, the existence of Philadelphia (Ph) chromosome positive progenitors with a silent BCR-ABL gene has been reported, thus raising concerns on the use of RT-PCR for detecting BCR-ABL positive progenitors. In order to investigate this issue further, colonies (n = 204) generated by mononuclear (MNC) or CD34+ CML cells were individually harvested, divided into two aliquots and analyzed both at the cytogenetic level to detect the Ph chromosome, and the molecular level to detect BCR-ABL transcripts. The mean (+/- s.d.) percentages of colonies analyzable by either SCK or RT-PCR were 74 +/- 16% and 86 +/- 16%, respectively. A significant percentage of colonies (67 +/- 19%) could be successfully analyzed by both SCK and RT-PCR. Although the majority of these colonies (97 +/- 5%) were Ph-positive and BCR-ABL-positive, a negligible percentage (4%) of progenitors were Ph-positive but BCR-ABL-negative. In order to test the influence of colony size on the outcome of molecular analysis, the efficiency of our RT-PCR assay in detecting BCR-ABL transcripts was investigated by means of experiments in which the number of cells used to start RNA extraction was serially reduced. These experiments showed that at least 150 cells were necessary to achieve a reproducible amplification of BCR-ABL transcripts. By correlating the size of harvested colonies with the outcome of molecular analysis, it was evident that BCR-ABL-negative but Ph-positive colonies represented false negative results occurring when a number of leukemic cells below the detection limit of our RT-PCR assay was analyzed. In conclusion, our data demonstrate that individual CML colonies grown in semisolid culture assays can be indifferently analyzed by SCK or RT-PCR, and support an extensive use of a carefully standardized RT-PCR assay to estimate the leukemic burden within samples which have been purged and selected in vitro.
- Published
- 1998
36. A prospective comparison between treatment with phlebotomy alone and with interferon-alpha in patients with polycythemia vera
- Author
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Mf Martelli, A Vecchi, Pietro Leoni, L Bensi, C Messora, Serena Rupoli, G. Ucci, Antonio Tabilio, Marina Liberati, Stefano Sacchi, Fausto Grignani, P Tartoni, Franca Falzetti, and A. Riccardi
- Subjects
Adult ,Male ,medicine.medical_specialty ,Alpha interferon ,Hematocrit ,Gastroenterology ,Polycythemia vera ,White blood cell ,Internal medicine ,ESSENTIAL THROMBOCYTHEMIA ,medicine ,Humans ,Prospective Studies ,Author Keywords:ALPHA-IFN ,PHLEBOTOMY ,Polycythemia Vera ,Interferon alfa ,Aged ,Bloodletting ,Chromosome Aberrations ,Hematology ,medicine.diagnostic_test ,business.industry ,Interferon-alpha ,General Medicine ,Middle Aged ,Phlebotomy ,medicine.disease ,Combined Modality Therapy ,Crossover study ,POLYCYTHEMIA VERA KeyWords Plus:MEGAKARYOBLASTIC TRANSFORMATION ,medicine.anatomical_structure ,Splenomegaly ,Immunology ,Female ,business ,medicine.drug - Abstract
Interferon alpha (alpha-IFN) is increasingly used for the treatment of patients affected by polycythemia vera (PV). As prior studies are difficult to interpret in view of the lack of appropriate controls, we undertook a randomized comparison of lymphoblastoid alpha-IFN (alpha n-1 IFN) treatment against venesection treatment alone. In a crossover trial, we treated 22 PV patients alternatively for 5 months each with 3 MU/day sc of alpha n-1 IFN and phlebotomy alone. During IFN treatment, red blood cell count and hematocrit level were well controlled in both trial groups, reducing or eliminating the need for phlebotomy in all patients; furthermore, platelet number and white blood cell count declined during alpha-IFN therapy. In addition, the number of symptomatic patients was greatly reduced, and in six patients a reduction in splenic size was observed. Finally, the only patient with chromosomal abnormalities showed a complete cytogenetic conversion after 5 months of alpha-IFN therapy. Thus, for the first time, our results provide the unequivocal demonstration that alpha-IFN is superior to phlebotomy in controlling the pathologic expansion of erythroid elements and all the clinical aspects of this disease.
- Published
- 1994
37. Adhesion Molecule Expression on B-Cells from Acute and Chronic Lymphoid Leukemias
- Author
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G. Cerruti, Luciana Annino, G. De Rossi, Francesca Romana Mauro, Daniela Zarcone, Antonio Tabilio, C. E. Grossit, Claudya Tenca, and Anna Favre
- Subjects
Male ,Cancer Research ,Stromal cell ,Fluorescent Antibody Technique ,Cell–cell interaction ,Humans ,Lymphocyte homing receptor ,Cell adhesion ,B-Lymphocytes ,CD40 ,biology ,Cell adhesion molecule ,Infant, Newborn ,Hematology ,Middle Aged ,Flow Cytometry ,Burkitt Lymphoma ,Leukemia, Lymphocytic, Chronic, B-Cell ,Cell biology ,Phenotype ,Oncology ,biology.protein ,Female ,Neural cell adhesion molecule ,Cell Adhesion Molecules ,Homing (hematopoietic) - Abstract
Adhesion molecule expression on acute and chronic lymphoid leukemia cells of B lineage (B-ALL and B-CLL) may subserve several functions. Adhesion of leukemic cells to endothelial cells and to extracellular matrix components is relevant to homing, trafficking and spread of the malignant cells, and thus to clinical presentation, course and disease prognosis. Adhesive interactions between malignant cells and accessory cells, particularly stromal cells in the bone marrow environment, may support growth of the malignant cells via cytokine-delivered messages. They may also deliver signals that prevent or trigger programmed cell death of tumor cells. Here we review data on the adhesive phenotype of leukemic blasts from pro-B (CALLA +) ALL and of cells from B-CLL cases. We show that expression of certain adhesion molecules may help define disease subsets with distinctive clinical and prognostic features. One adhesion molecule, the lymphocyte homing receptor CD44, allows definition of two groups of B-CLL patients with significantly different survival.
- Published
- 1994
38. Expression of the FMS/KIT-like gene FLT3 in human acute leukemias of the myeloid and lymphoid lineages
- Author
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Sylvie Marchetto, Olivier Rosnet, Marie-Josèphe Pébusque, Daniel Birnbaum, Marianne Courcoul, Patrice Mannoni, Françoise Birg, Florence Bardin, and Antonio Tabilio
- Subjects
Myeloid ,T-Lymphocytes ,Immunology ,Gene Expression ,Biology ,Biochemistry ,Monocytes ,fluids and secretions ,Proto-Oncogene Proteins ,hemic and lymphatic diseases ,Gene expression ,medicine ,Humans ,RNA, Messenger ,Northern blot ,Progenitor cell ,B-Lymphocytes ,Chromosomes, Human, Pair 13 ,Nucleic Acid Hybridization ,Myeloid leukemia ,hemic and immune systems ,Genes, fms ,Cell Biology ,Hematology ,Precursor Cell Lymphoblastic Leukemia-Lymphoma ,Blotting, Northern ,medicine.disease ,Blot ,Leukemia, Myeloid, Acute ,Proto-Oncogene Proteins c-kit ,Leukemia ,medicine.anatomical_structure ,embryonic structures ,Cancer research ,Chromosome Deletion ,DNA Probes ,Granulocytes - Abstract
FLT3, a receptor belonging to the FMS/KIT family and localized to 13q12, could play a role in the biology of early hematopoietic progenitor cells. Because FMS and KIT are expressed in both normal progenitors and myeloid leukemias, we looked for FLT3 expression in fresh human leukemic cells using Northern blot analysis. High levels of FLT3 expression were detected in 92% of the cases of acute myeloid leukemia (AML) tested, ranging from the M1 to the M5 stages of differentiation assessed in the French-American-British classification. Immature (MO) AML cells, biphenotypic leukemias, and AML with megakaryocytic differentiation (M7 subtype) also expressed the FLT3 transcript. FLT3 was also expressed at high levels in acute lymphoid leukemias of T and B origins. Finally, it was not expressed in chronic myeloid leukemias in chronic phase, whereas it was expressed in most blast crisis samples. This pattern of expression of FLT3 contrasts with the expression of FMS and KIT restricted to myeloid leukemias, and suggests that the FLT3 product could play a role in the expansion of the leukemic blasts of both the myeloid and lymphoid lineages.
- Published
- 1992
39. Immunomagnetic isolation of CD4+CD25+FoxP3+ natural T regulatory lymphocytes for clinical applications
- Author
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Elisabetta Bonifacio, M Di Ianni, Franca Falzetti, R. Iacucci Ostini, Debora Cecchini, Lorenzo Moretti, Massimo F. Martelli, B. Del Papa, Antonio Tabilio, Tiziana Zei, L. Fontana, G. Tagliapietra, and C. Maldini
- Subjects
Adult ,CD4-Positive T-Lymphocytes ,Translational Studies ,medicine.drug_class ,Immunology ,Population ,chemical and pharmacologic phenomena ,Biology ,Monoclonal antibody ,Immunophenotyping ,Interleukin-7 Receptor alpha Subunit ,Interleukin-5 Receptor alpha Subunit ,medicine ,Immunology and Allergy ,Humans ,IL-2 receptor ,Leukapheresis ,education ,Interleukin-7 receptor ,education.field_of_study ,Immunomagnetic Separation ,Reverse Transcriptase Polymerase Chain Reaction ,Spectral Karyotyping ,Interleukin-2 Receptor alpha Subunit ,FOXP3 ,hemic and immune systems ,Forkhead Transcription Factors ,Middle Aged ,medicine.disease ,Graft-versus-host disease ,Phenotype ,CD8 - Abstract
Although CD4(+)/CD25(+) T regulatory cells (T(regs)) are a potentially powerful tool in bone marrow transplantation, a prerequisite for clinical use is a cell-separation strategy complying with good manufacturing practice guidelines. We isolated T(regs) from standard leukapheresis products using double-negative selection (anti-CD8 and anti-CD19 monoclonal antibodies) followed by positive selection (anti-CD25 monoclonal antibody). The final cell fraction (CD4(+)/CD25(+)) showed a mean purity of 93.6% +/- 1.1. Recovery efficiency was 81.52% +/- 7.4. The CD4(+)/CD25(+bright) cells were 28.4% +/- 6.8. The CD4(+)/CD25(+) fraction contained a mean of 51.9% +/- 15.1 FoxP3 cells and a mean of 18.9% +/- 11.5 CD127 cells. Increased FoxP3 and depleted CD127 mRNAs in CD4(+)CD25(+)FoxP3(+) cells were in line with flow cytometric results. In Vbeta spectratyping the complexity scores of CD4(+)/CD25(+) cells and CD4(+)/CD25(-) cells were not significantly different, indicating that T(regs) had a broad T cell receptor repertoire. The inhibition assay showed that CD4(+)/CD25(+) cells inhibited CD4(+)/CD25(-) cells in a dose-dependent manner (mean inhibition percentages: 72.4 +/- 8.9 [ratio of T responder (T(resp)) to T(regs), 1:2]; 60.8% +/- 20.5 (ratio of T(resp) to T(regs), 1:1); 25.6 +/- 19.6 (ratio of T(resp) to T(regs), 1:0.1)). Our study shows that negative/positive T(reg) selection, performed using the CliniMACS device and reagents, enriches significantly CD4(+)CD25(+)FoxP3(+) cells endowed with immunosuppressive capacities. The CD4(+)CD25(+)FoxP3(+) population is a source of natural T(reg) cells that are depleted of CD8(+) and CD4(+)/CD25(-) reacting clones which are potentially responsible for triggering graft-versus-host disease (GvHD). Cells isolated by means of this approach might be used in allogeneic haematopoietic cell transplantation to facilitate engraftment and reduce the incidence and severity of GvHD without abrogating the potential graft-versus-tumour effect.
- Published
- 2009
40. Role of reactive oxygen species as signal molecules in the pre-commitment phase of adult stem cells
- Author
-
Claudia, Piccoli, Annamaria, D'Aprile, Rosella, Scrima, Maria, Ripoli, Domenico, Boffoli, Antonio, Tabilio, and Nazzareno, Capitanio
- Subjects
Adult Stem Cells ,Humans ,NADPH Oxidases ,Hematopoietic Stem Cells ,Reactive Oxygen Species ,Models, Biological ,Oxidation-Reduction ,Cell Proliferation ,Mitochondria - Abstract
This mini-review summarizes evidence, provided by our group, relevant to the understanding of how redox signalling may control the fate of adult hematopoietic stem/progenitor cells (HSPCs). In particular it is shown that bone marrow-derived human HSPC are endowed with a composite panel of constitutively active NADPH-oxidases (NOXs) comprising the cell membrane-localized catalytic subunits of the NOX1, NOX2 and NOX4 isoforms. It is proposed that the coordinated activity of the NOX isoforms in HSPCs function as environmental oxygen sensor and generate low level of ROS, which likely serve as second messengers. The pro-oxidant setting, entering into play when HSPCs leave the hypoxic bone marrow niche, would enable them to be more responsive to proliferative/differentiative stimuli. Moreover it is suggested that enhanced ROS elicit mitochondrial "differentiation" in a pre-commitment phase needed to match the bioenergetic request in the oncoming proliferation/differentiation process.
- Published
- 2009
41. Lambda-like and V pre-B genes expression: an early B-lineage marker of human leukemias
- Author
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David Bossy, Claudine Schiff, Patrice Mannoni, Antonio Tabilio, Jean Gabert, Michèle Milili, Franca Falzetti, and Michel Fougereau
- Subjects
Genetic Markers ,Transcription, Genetic ,Molecular Sequence Data ,Immunology ,Locus (genetics) ,Biology ,Polymerase Chain Reaction ,Biochemistry ,Germline ,Immunophenotyping ,Antigens, CD ,Gene expression ,Leukemia, B-Cell ,Humans ,RNA, Neoplasm ,Northern blot ,Gene Rearrangement, B-Lymphocyte ,Gene ,Genetics ,Regulation of gene expression ,Base Sequence ,Genes, Immunoglobulin ,Gene Expression Regulation, Leukemic ,DNA, Neoplasm ,Cell Biology ,Hematology ,Gene rearrangement ,Molecular biology ,Protein Biosynthesis - Abstract
V pre-B and lambda-like genes are selectively expressed in human pre-B cells and encode polypeptide chains that associate in a mu-pseudolight chain complex that may regulate some crucial steps of early B-cell differentiation. We have followed by polymerase chain reaction and Northern blot analysis the expression of these “pre-B-specific” genes in correlation with the status (rearranged v germline) of Ig gene loci (H, kappa, lambda) in a panel of 32 leukemias pertaining mostly to the B lineage and including a number of ambiguously characterized samples. All cells that had rearranged the H locus only expressed V pre-B and lambda-like transcripts, in agreement with a pre-B status. In this group, some biphenotypic leukemias (mostly My/B) might, in fact, be already engaged in the B lineage. Rearrangement of V kappa or V lambda loci correlated with the disappearance of the pre-B gene products. In a pre-B acute lymphoblastic leukemia cell line that was induced to mature to the B-cell stage in culture upon kappa gene rearrangement, the mu- pseudolight chain complex was actually replaced by the classical mu- kappa molecule. Finally, V pre-B and lambda-like genes were found expressed in two leukemic cells that had retained all Ig loci in germline configuration. This finding raises the possibility of having an early pro-B progenitor in which V pre-B and lambda-like products associate with a H chain surrogate in a complex that would trigger an early event of B-cell differentiation such as the H locus rearrangements.
- Published
- 1991
42. A randomised clinical trial comparing idarubicin and cytarabine to daunorubicin and cytarabine in the treatment of acute non-lymphoid leukaemia
- Author
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Nicola Di Pietro, Roberto Latagliata, Luigi Resegotti, Maria Concetta Petti, Francesco Nobile, Franco Mandelli, Saverio Ladogana, Nicola Petti, Claudio Malleo, Emanuela Falconi, Bruno Rotoli, Giorgina Specchia, Fabrizio Ganzina, Enrico Geraci, Alfredo Ardia, Francesco Di Raimondo, and Antonio Tabilio
- Subjects
medicine.medical_specialty ,Pediatrics ,Myeloid ,Performance status ,business.industry ,Daunorubicin ,food and beverages ,biochemical phenomena, metabolism, and nutrition ,medicine.disease ,Chemotherapy regimen ,Gastroenterology ,carbohydrates (lipids) ,Leukemia ,medicine.anatomical_structure ,Oncology ,Maintenance therapy ,hemic and lymphatic diseases ,Internal medicine ,medicine ,Cytarabine ,Idarubicin ,lipids (amino acids, peptides, and proteins) ,business ,medicine.drug - Abstract
255 patients with acute non-lymphoid leukaemia (ANLL), observed between October 1984 and June 1987, entered a chemotherapy regimen consisting of induction therapy with cytarabine in combination with idarubicin (IDA/ARA) or daunorubicin (DNR/ARA), followed by consolidation with four courses of IDA+ARA plus 6-thioguanine (6-TG) or DNR+ARA+6-TG and a 6 month maintenance therapy with 6-TG and ARA. The median age was 62 years (range 55–78 years) and 33 were aged more than 70 years. The treatment groups were comparable for median age, FAB type, performance status and initial blood counts. 249 patients were randomised, 124 to the IDA/ARA arm and 125 to the DNR/ARA arm. Complete remission was achieved in 50 patients (40%) on the IDA/ARA treatment program and 49 patients (39%) on DNR/ARA. No definite differences were found between patients receiving IDA/ARA and those treated with DNR/ARA as far as complete response (CR), overall survival, failure free and relapse free survival are concerned. 74% of the complete responders in the IDA/ARA arm and 51% in the DNR/ARA arm achieved CR after a single course of treatment. Resistant leukaemia was observed in 13.7% of the patients in the IDA/ARA arm and in 31.2% in the DNR/ARA one, whereas hypoplastic death occurred in 29% and 14.4%, respectively. In conclusion, our data failed to show any advantage of idarubicin over daunorubicin even though there is some evidence that IDA, despite the higher toxicity, is more rapid in eradicating leukaemia as proved by the higher CR rate obtained after one course of induction.
- Published
- 1991
43. Activation of the granulocyte-monocyte colony stimulating factor gene in acute myeloid leukaemia cells is not related to gene rearrangement
- Author
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Flavio Falcinelli, Marina Onorato, Raffaella Ciurnelli, Patrice Mannoni, Jean Gabert, Franca Falzetti, Massimo F. Martelli, and Antonio Tabilio
- Subjects
Gene Rearrangement ,Macrophage colony-stimulating factor ,Cancer Research ,Monocyte ,Structural gene ,Granulocyte-Macrophage Colony-Stimulating Factor ,Hematology ,Gene rearrangement ,Biology ,Colony-stimulating factor ,Gene Expression Regulation, Neoplastic ,Leukemia, Myeloid, Acute ,Granulocyte macrophage colony-stimulating factor ,medicine.anatomical_structure ,Genes ,Oncology ,hemic and lymphatic diseases ,Immunology ,medicine ,Humans ,Colony-Stimulating Factor Gene ,RNA, Messenger ,RNA, Neoplasm ,Autocrine signalling ,medicine.drug - Abstract
Several reports have documented that leukaemic blasts produce a number of cytokines among them the granulocyte-monocyte colony stimulating factor (GM-CSF). We analysed the structure of the gene that codes for GM-CSF in 44 acute myeloid leukaemia (AML) cases in an attempt to establish whether the autocrine production of GM-CSF was due to a structural gene alteration. No structural alteration was detected in the GM-CSF gene in any of the 44 cases studied. We, therefore, conclude that the autocrine production of GM-CSF by leukaemia blasts is not dependent on gene rearrangement.
- Published
- 1991
44. Constitutively activated Notch signaling is involved in survival and apoptosis resistance of B-CLL cells
- Author
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Pierfrancesco Marconi, Rita Sabatini, Emanuela Rosati, Isabella Screpanti, Giuliana Rampino, Katia Fettucciari, Mauro Di Ianni, Antonio Tabilio, Stefano Coaccioli, and Andrea Bartoli
- Subjects
Apoptosis Inhibitor ,Cell Survival ,Immunology ,Notch signaling pathway ,Down-Regulation ,Apoptosis ,Biology ,Inhibitor of apoptosis ,Biochemistry ,Inhibitor of Apoptosis Proteins ,Downregulation and upregulation ,hemic and lymphatic diseases ,Tumor Cells, Cultured ,Humans ,Serrate-Jagged Proteins ,RNA, Small Interfering ,B-Lymphocytes ,Receptors, Notch ,Calcium-Binding Proteins ,NF-kappa B ,Membrane Proteins ,Cell Biology ,Hematology ,Leukemia, Lymphocytic, Chronic, B-Cell ,XIAP ,Cell biology ,Hes3 signaling axis ,Intercellular Signaling Peptides and Proteins ,Signal transduction ,Jagged-2 Protein ,Jagged-1 Protein ,Signal Transduction - Abstract
Notch signaling is involved in tumorigenesis, but its role in B–chronic lymphocytic leukemia (B-CLL) pathogenesis is not completely defined. This study examined the expression and activation of Notch receptors in B-CLL cells and the role of Notch signaling in sustaining the survival of these cells. Our results show that B-CLL cells but not normal B cells constitutively express Notch1 and Notch2 proteins as well as their ligands Jagged1 and Jagged2. Notch signaling is constitutively activated in B-CLL cells, and its activation is further increased in B-CLL cells, which resist spontaneous apoptosis after 24-hour ex vivo culture. Notch stimulation by a soluble Jagged1 ligand increases B-CLL cell survival and is accompanied by increased nuclear factor–kappa B (NF-κB) activity and cellular inhibitor of apoptosis protein 2 (c-IAP2) and X-linked inhibitor of apoptosis protein (XIAP) expression. In contrast, Notch-signaling inhibition by the γ-secretase inhibitor I (GSI; z-Leu-Leu-Nle-CHO) and the specific Notch2 down-regulation by small-interfering RNA accelerate spontaneous B-CLL cell apoptosis. Apoptotic activity of GSI is accompanied by reduction of NF-κB activity and c-IAP2 and XIAP expression. Overall, our findings show that Notch signaling plays a critical role in B-CLL cell survival and apoptosis resistance and suggest that it could be a novel potential therapeutic target.
- Published
- 2008
45. Evidence of jak2 val617phe positive essential thrombocythemia with splanchnic thrombosis during estroprogestinic treatment
- Author
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Marianna Colagrande, Guglielmo Mariani, Antonio Tabilio, Mauro Di Ianni, Mariasanta Napolitano, Mario Lapecorella, Alessandro Lucchesi, Lapecorella M., Tabilio A., Lucchesi A., Napolitano M., Colagrande M., Di Ianni M., Mariani G., Lapecorella, M, Tabilio, A, Lucchesi,A, Napolitano, M, Colagrande, M, Di Ianni,M, and Mariani, G
- Subjects
Adult ,medicine.medical_specialty ,Mutation, Missense ,Oral contraceptive ,Essential thrombocythemia ,Gastroenterology ,Contraceptives, Oral, Hormonal ,Pathogenesis ,Mesenteric Veins ,Portal thrombosis ,Myeloproliferative Disorders ,Internal medicine ,medicine ,Humans ,Platelet ,Splanchnic Circulation ,Janus kinase 2 ,biology ,essential thrombocythemia, Janus kinase 2, oral contraceptives, portal thrombosis ,Kinase ,Vascular disease ,business.industry ,Thrombosis ,Hematology ,General Medicine ,medicine.disease ,Thrombocytopenia ,Endocrinology ,biology.protein ,Female ,business - Abstract
The discovery of the Janus kinase 2 Val617Phe mutation has brought new insights into the development of myeloproliferative disorders; however, the pathogenesis of essential thrombocythemia and its related thrombotic complications has not been completely understood. Although the Janus kinase 2 Val617Phe mutation confirms the initially suspected clonal character of the disease, factors influencing clonal transformation and expansion in the bone marrow have not been fully detected. Furthermore, patients affected by essential thrombocythemia who are carriers of the Janus kinase 2 Val617Phe mutation show a higher incidence of venous thromboembolism both before, and at the time of diagnosis, compared with noncarriers, and recent evidence of splanchnic and cerebral vein thrombosis in carriers of the Janus kinase 2 Val617Phe mutation has been reported. The intake of oral contraceptives is a strong and independent risk factor for venous thromboembolism. In addition, in-vitro tests showed both an altered primary haemostatic plug formation and enhanced platelet aggregation in patients taking such drugs. Little is known, though, about the influence of steroid hormones on both megakaryopoiesis and platelet function in patients with the Janus kinase 2 Val617Phe mutation. Herewith, we report the case of a 30-year-old woman who took a third generation oral contraceptive for 5 months and developed an essential thrombocythemia with splenog-portal axis and superior mesenteric vein thrombosis. She was found to carry the kinase gene Janus kinase 2 mutation. © 2008 Wolters Kluwer Health | Lippincott Williams & Wilkins.
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- 2008
46. Eubacterium plautii infection in a kidney transplant recipient: a noteworthy case of pleural effusion and fever
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Giuseppe, Orlando, Francesco, Pisani, Paola, Mastrantonio, Luigi, Bonanni, Pierpaolo, Di Cocco, Maurizio, D'Angelo, Antonio, Tabilio, and Antonio, Famulari
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Adult ,Male ,Pleural Effusion ,Postoperative Complications ,Fever ,Eubacterium ,Humans ,Kidney Diseases ,Kidney Transplantation ,Gram-Positive Bacterial Infections - Abstract
We report a noteworthy case of Eubacterium plautii infection after kidney transplantation. Our 33-yr-old transplant recipient received standard care; his post-transplant course was uneventful. However, on day 44 he underwent an emergency laparotomy for perforation of the ileum. He was initially treated with ceftazidime, fluconazole and metronidazole, but his fever persisted, so he was switched to meropenem and vancocin. We could not find any cause for his infection. On day 70, his temperature normalized. On day 75, he developed severe leukopenia (280 cell/mL). His cytomegalovirus-DNA test result was negative, so all immunosuppressants, except for prednisone, were stopped; instead, antibiotic prophylaxis was started, using caspofungin, trimethoprim-sulfamethoxazole and ciprofloxacin. On day 83, he underwent percutaneous drainage of massive left pleural effusion. We repeatedly cultured the pleural liquid, but it was not till three wk later that we were finally able to identify the causative organism. We hypothesize that the microorganism - which normally resides on the surface of the intestinal lumen - entered the bloodstream via bacterial translocation, eventually colonizing the pleurae. This translocation was favored by our patient poor clinical condition, his immunosuppressive treatment and his heavy antibiotherapy. Our experience highlights the need for wiser use of antibiotics in transplant recipients.
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- 2008
47. Topological organization of NADPH-oxidase in haematopoietic stem cell membrane: preliminary study by fluorescence near-field optical microscopy
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M.C. Frassanito, Domenico Boffoli, Claudia Piccoli, Nazzareno Capitanio, Vito Capozzi, and Antonio Tabilio
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chemistry.chemical_classification ,Reactive oxygen species ,Histology ,NADPH oxidase ,Microscopy, Confocal ,biology ,Cell growth ,Cell Membrane ,NADPH Oxidases ,Microscopy, Scanning Probe ,Hematopoietic Stem Cells ,Immunohistochemistry ,Pathology and Forensic Medicine ,Cell biology ,Haematopoiesis ,Membrane ,chemistry ,Membrane protein ,Microscopy, Fluorescence ,biology.protein ,Humans ,Stem cell ,NOx - Abstract
Summary The aim of this study was to characterize the local distribution and organization of the plasma membrane NADPH-oxidase (NOX) in human haematopoietic stem cell (HSC) by means of the fluorescence scanning near-field optical microscopy approach. The presence of NOX in haematopoietic stem cells is thought to have a functional role as O2 sensor and/or as low-level reactive oxygen species (ROS) producer to be used as redox messenger for controlling cell growth and differentiation. Given the harmful potential of ROS, a fine-tuning of NOX activity is needed. The high resolution imaging of haematopoietic stem cell membrane obtained in this study combined with the immunodetection of NOX indicates for this the occurrence of a cluster-organized structure. These membrane ‘rafts’-like micro-compartments may constitute localized protein aggregates whereby the assembly/activation of the NOX components are functionally integrated with upstream factors constituting signal-transduction platforms.
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- 2008
48. Efalizumab-induced immune thrombocytopenia during retreatment
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Gino Coletti, Ketty Peris, Antonio Tabilio, and Maria Concetta Fargnoli
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Anticorps monoclonal ,Antibodies monoclonal ,business.industry ,Psoriasis ,Efalizumab ,Immunology ,medicine ,Dermatology ,medicine.disease ,business ,Immune thrombocytopenia ,medicine.drug - Published
- 2008
49. Mesenchymal cells recruit and regulate T regulatory cells
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Paolo Sportoletti, Maria De Ioanni, Lorenzo Moretti, Debora Cecchini, Franca Falzetti, Elisabetta Bonifacio, Mauro Di Ianni, Antonio Tabilio, and Beatrice Del Papa
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Adult ,Cancer Research ,CD3 Complex ,T cell ,CD3 ,Down-Regulation ,chemical and pharmacologic phenomena ,Biology ,T-Lymphocytes, Regulatory ,Flow cytometry ,Interleukin-7 Receptor alpha Subunit ,Immune system ,Reference Values ,Genetics ,medicine ,Humans ,IL-2 receptor ,Lymphocytes ,Interleukin-7 receptor ,Molecular Biology ,Cells, Cultured ,medicine.diagnostic_test ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Expression Profiling ,Mesenchymal stem cell ,Interleukin-2 Receptor alpha Subunit ,FOXP3 ,hemic and immune systems ,Forkhead Transcription Factors ,Mesenchymal Stem Cells ,Cell Biology ,Hematology ,Flow Cytometry ,Molecular biology ,Coculture Techniques ,medicine.anatomical_structure ,CD4 Antigens ,biology.protein ,Leukocyte Common Antigens - Abstract
Objective Despite much investigation into T regulatory cells (Tregs), little is known about the mechanism controlling their recruitment and function. Because multipotent mesenchymal stromal cells (MSCs) exert an immune regulatory function and suppress T-cell proliferation, this in vitro study investigated their role in Treg recruitment and function. Materials and Methods Human MSCs and different T cell populations (CD3 + , CD3 + /CD45RA + , CD3 + /CD45RO + , CD4 + /CD25 + , CD4 + /CD25 + /CD45RO + , CD4 + /CD25 + /CD45RA + ) from healthy donors were cocultured for up to 15 days. Harvested lymphocytes were analyzed by flow cytometry and FoxP3 and CD127 expressions were measured by real-time polymerase chain reaction. Their regulatory activity was assessed. Results We demonstrate MSC recruit Tregs from a fraction of CD3 + and from immunoselected CD3 + /CD45RA + and CD3 + /CD45RO + fractions. After culture with MSCs both immunoselected fractions registered increases in the CD4 + /CD25 bright /FoxP3 subset and CD127 expression was downregulated. When purified Treg populations (CD4/CD25 + , CD4/CD25 + /CD45RA + , and CD4/CD25 + /CD45RO + ) are used in MSC cocultures, they maintain FoxP3 expression and CD127 expression is downregulated. Treg suppressive capacity was maintained in Treg populations that were layered on MSC for up to 15 days while control Tregs lost all suppressive activity after 5 days culture. Conclusions In conclusion, our study demonstrates that MSCs recruit, regulate, and maintain T-regulatory phenotype and function over time.
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- 2008
50. Transformation by retroviral vectors of bone marrow-derived mesenchymal cells induces mitochondria-dependent cAMP-sensitive reactive oxygen species production
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Maria Paola Martelli, Nazzareno Capitanio, Annamaria D'Aprile, Antonio Tabilio, Beatrice Del Papa, Maria Ripoli, Rosella Scrima, Claudia Piccoli, Domenico Boffoli, Mauro Di Ianni, Giuseppe Servillo, Claudio Ligas, and Giovanni Quarato
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Stromal cell ,Genetic enhancement ,Cell Respiration ,Respiratory chain ,Bone Marrow Cells ,Biology ,Viral vector ,Cell Line ,Mesoderm ,Transduction (genetics) ,Transduction, Genetic ,Cyclic AMP ,Humans ,Electron Transport Complex I ,Interleukin-7 ,Mesenchymal stem cell ,Lentivirus ,Cell Biology ,Transfection ,Cell Transformation, Viral ,Molecular biology ,Cyclic AMP-Dependent Protein Kinases ,Mitochondria ,Cell culture ,Molecular Medicine ,Moloney murine leukemia virus ,Stromal Cells ,Reactive Oxygen Species ,Oxidation-Reduction ,Developmental Biology - Abstract
Retroviral vectors are used in human gene therapy trials to stably introduce therapeutic genes in the genome of patients' cells. Their applicability, however, is frustrated by the limited viability of transformed cells and/or by risks linked to selection of oncogene-mutated clones. The reasons for these drawbacks are not yet completely understood. In this study, we show that LXSN-NeoR gene/interleukin-7-engineered mesenchymal stromal cells exhibited a marked enhancement of reactive oxygen species production compared with untransfected cells. This effect resulted to be independent on the product of the gene carried by the retroviral vehicle as it was reproducible in cells transfected with the empty vector alone. Stable transfection of mesenchymal stromal cells with the different retroviral vectors pBabe-puro and PINCO-puro and the lentiviral vector pSico PGK-puro caused similar redox imbalance, unveiling a phenomenon of more general impact. The enhanced production of reactive oxygen species over the basal level was attributable to mitochondrial dysfunction and brought back to altered activity of the NADH-CoQ oxidoreductase (complex I) of the respiratory chain. The oxidative stress in transfected mesenchymal stem cells was completely reversed by treatment with a cAMP analog, thus pointing to alteration in the protein kinase A-dependent signaling pathway of the host cell. Transfection of mesenchymal stromal cells with a PINCO-parental vector harboring the green fluorescent protein gene as selection marker in place of the puromycin-resistance gene resulted in no alteration of the redox phenotype. These novel findings provide insights and caveats to the applicability of cell- or gene-based therapies and indicate possible intervention to improve them. Disclosure of potential conflicts of interest is found at the end of this article.
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- 2008
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