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Successful use of donor lymphocyte infusions in post-BMT relapse of RAEB (monosomy 7)
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8. Advanced MDS: Hematopoietic Stem Cell Transplantation SUCCESSFUL USE OF DONOR LYMPHOCYTE INFUSIONS IN POST-BMT RELAPSE OF RAEB (MONOSOMY 7)
R. Skinner1 *, M. Velangi2, N. Bown 3. IPaediatric Oncology/BMT, Newcastle upon Tyne Hospitals NHS Trust, Newcastle upon Tyne, United Kingdom," 2Paediatric Haematology, Newcastle upon Tyne Hospitals NHS Trust, Newcastle upon Tyne, United Kingdom," 3Northern Genetics Service, Newcastle upon Tyne Hospitals NHS Trust, Newcastle upon Tyne, United Kingdom *E-mail: Roderick. [email protected] Introduction: Haemopoietic stem cell transplantation (HSCT)
is the only curative treatment for advanced myelodysplasia in children, but there is little information about the use of donor lymphocyte infusions (DLI) in post-HSCT relapse. Case Report: A 5 year old boy presented with pancytopenia due to RAEB (marrow 15% blasts). Marrow cytogenetics revealed monosomy 7. He received 4 courses of intensive "AM]A' chemotherapy. Haematological recovery was slow, but repeat cytogenetics showed disappearance of monosomy 7. He underwent unrelated donor HLA-identical unmanipulated BMT, conditioned with busulphan (16 mg/kg), cyclophosphamide (120 mg/kg), melphalan (140mg/m2), rabbit ATG (2mg/kg/day, days - 5 to -2). PostBMT GvHD prophylaxis was cyclosporine A and short course methotrexate. Neutrophil engraftment occurred by day +20, platelets by day +29; full donor chimerism was demonstrated on day +39. CMV reactivation on day +47 was treated with ganciclovir and cutaneous acute GvHD (Grade I) on day +64 with low dose oral prednisolone. He remained well until 21 months postBMT when he was again pancytopenic. Bone marrow confirmed relapse of RAEB, monosomy 7 with further clonal evolution (trisomy 21) and mixed chimerism (MC). He received 3 doses (escalating dose schedule) of DLI over 3 months, but pancytopenia worsened (neutrophils <0.1• with evolving graft failure (severe marrow hypocellularity). He developed probable fungal pneumonia (nodules on CT scan, no organisms on BAL), treated with high dose liposomal amphotericin, itraconazole and GM-CSE Cutaneous acute GvHD (Grade II) occurred after the last DLI, resolving with topical steroids only. DLI-induced marrow aplasia, on a background of progressive relapse, persisted 2 months after the last DLI. Plans were made for an unconditioned second HSCT, using PBSC, but over the next month his general condition and blood count improved, with increased marrow cellularity, blasts <1%, no monosomy 7 and 100% donor chimerism. Despite slow immunological reconstitution his pneumonia cleared clinically and radiologically. He remains well, with normal blood count and full donor chimerism, 4.5 years later. Conclusion: DLI for post-BMT relapse of RAEB (monosomy 7) led to full recovery with prolonged disease-free survival, despite initial severe marrow hypocellularity and presumed fungal pneumonia. Improvement followed mild cutaneous GvHD. No chemotherapy was given, implying that a graft versus myelodysplasia effect was responsible.
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SINGLE CELL CLONING OF HUMAN, DONORDERIVED ANTI-LEUKEMIA T CELL LINES FOR IN VITRO SEPARATION OF GRAFT-VERSUSLEUKEMIA EFFECT FROM GRAFT-VERSUS-HOST REACTION
D. Montagna 1'2, L. Daudt 2, F. Locatelli2., E. Montini2, I. Turin2, D. Lisini2, G. Giorgiani2, M.E. Bernardo 2, R. Maccario 2.
1Department of Pediatrics, Laboratory of Immunology, University of Pavia, Pavia, Italy," 2Pediatric Hematology-Oncology Unit, IRCCS Policlinico San Matteo, University of Pavia, Pavia, Italy *E-mail: [email protected] In previous studies, we demonstrated the possibility of expanding in vitro polyclonal cytotoxic T-lymphocyte (CTL) lines directed against patient leukemia cells, using effector cells derived from their, both HLA-matched and HLA-mismatched, hematopoietic stem cell (HSC) donors. Some CTL lines, especially those derived from an HLA-disparate donor, displayed residual alloreactivity against patient non-malignant cells. In this study, we evaluated the possibility of separating in vitro CTLs with selective graftversus-leukemia (GVL) activity from those potentially involved in the development of graft-versus-host disease (GVHD), through single T-cell cloning of anti-leukemia polyclonal CTL lines. Five patients with acute leukemia (3 with acute myeloid leukemia, AML, in one case diagnosed in a child who had been previously treated for osteosarcoma, and 2 with acute lymphoblastic leukemia, ALL), given an allograft from either an HLA-identical sibling (2 children) or an HLA-partially matched relative (3 children), were included in the study. We demonstrated that CTLs expanded from a single T-cell clone (TCC), able to selectively kill leukemia blasts (LBs) and devoid of alloreactivity towards non-malignant cells, can be obtained from anti-leukemia alloreactive polyclonal CTL lines. Such TCCs expressed a wide repertoire of different T-cell receptor (TCR)-V~-families, mainly produced Interferon-7 and Interleukin-2, irrespective of CD8 or CD4 phenotype, and could be extensively expanded in vitro, without losing their peculiar functional features. The feasibility of our approach to in vitro separation of GVL from GVH reaction opens perspectives for using TCCs, selectively reactive towards LBs, for approaches of antileukemia adoptive immune therapy after HSC transplantation, in particular from HLA-mismatched donors.
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8. Advanced MDS: Hematopoietic Stem Cell Transplantation SUCCESSFUL USE OF DONOR LYMPHOCYTE INFUSIONS IN POST-BMT RELAPSE OF RAEB (MONOSOMY 7)
R. Skinner1 *, M. Velangi2, N. Bown 3. IPaediatric Oncology/BMT, Newcastle upon Tyne Hospitals NHS Trust, Newcastle upon Tyne, United Kingdom," 2Paediatric Haematology, Newcastle upon Tyne Hospitals NHS Trust, Newcastle upon Tyne, United Kingdom," 3Northern Genetics Service, Newcastle upon Tyne Hospitals NHS Trust, Newcastle upon Tyne, United Kingdom *E-mail: Roderick. [email protected] Introduction: Haemopoietic stem cell transplantation (HSCT)
is the only curative treatment for advanced myelodysplasia in children, but there is little information about the use of donor lymphocyte infusions (DLI) in post-HSCT relapse. Case Report: A 5 year old boy presented with pancytopenia due to RAEB (marrow 15% blasts). Marrow cytogenetics revealed monosomy 7. He received 4 courses of intensive "AM]A' chemotherapy. Haematological recovery was slow, but repeat cytogenetics showed disappearance of monosomy 7. He underwent unrelated donor HLA-identical unmanipulated BMT, conditioned with busulphan (16 mg/kg), cyclophosphamide (120 mg/kg), melphalan (140mg/m2), rabbit ATG (2mg/kg/day, days - 5 to -2). PostBMT GvHD prophylaxis was cyclosporine A and short course methotrexate. Neutrophil engraftment occurred by day +20, platelets by day +29; full donor chimerism was demonstrated on day +39. CMV reactivation on day +47 was treated with ganciclovir and cutaneous acute GvHD (Grade I) on day +64 with low dose oral prednisolone. He remained well until 21 months postBMT when he was again pancytopenic. Bone marrow confirmed relapse of RAEB, monosomy 7 with further clonal evolution (trisomy 21) and mixed chimerism (MC). He received 3 doses (escalating dose schedule) of DLI over 3 months, but pancytopenia worsened (neutrophils <0.1• with evolving graft failure (severe marrow hypocellularity). He developed probable fungal pneumonia (nodules on CT scan, no organisms on BAL), treated with high dose liposomal amphotericin, itraconazole and GM-CSE Cutaneous acute GvHD (Grade II) occurred after the last DLI, resolving with topical steroids only. DLI-induced marrow aplasia, on a background of progressive relapse, persisted 2 months after the last DLI. Plans were made for an unconditioned second HSCT, using PBSC, but over the next month his general condition and blood count improved, with increased marrow cellularity, blasts <1%, no monosomy 7 and 100% donor chimerism. Despite slow immunological reconstitution his pneumonia cleared clinically and radiologically. He remains well, with normal blood count and full donor chimerism, 4.5 years later. Conclusion: DLI for post-BMT relapse of RAEB (monosomy 7) led to full recovery with prolonged disease-free survival, despite initial severe marrow hypocellularity and presumed fungal pneumonia. Improvement followed mild cutaneous GvHD. No chemotherapy was given, implying that a graft versus myelodysplasia effect was responsible.
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]
SINGLE CELL CLONING OF HUMAN, DONORDERIVED ANTI-LEUKEMIA T CELL LINES FOR IN VITRO SEPARATION OF GRAFT-VERSUSLEUKEMIA EFFECT FROM GRAFT-VERSUS-HOST REACTION
D. Montagna 1'2, L. Daudt 2, F. Locatelli2., E. Montini2, I. Turin2, D. Lisini2, G. Giorgiani2, M.E. Bernardo 2, R. Maccario 2.
1Department of Pediatrics, Laboratory of Immunology, University of Pavia, Pavia, Italy," 2Pediatric Hematology-Oncology Unit, IRCCS Policlinico San Matteo, University of Pavia, Pavia, Italy *E-mail: [email protected] In previous studies, we demonstrated the possibility of expanding in vitro polyclonal cytotoxic T-lymphocyte (CTL) lines directed against patient leukemia cells, using effector cells derived from their, both HLA-matched and HLA-mismatched, hematopoietic stem cell (HSC) donors. Some CTL lines, especially those derived from an HLA-disparate donor, displayed residual alloreactivity against patient non-malignant cells. In this study, we evaluated the possibility of separating in vitro CTLs with selective graftversus-leukemia (GVL) activity from those potentially involved in the development of graft-versus-host disease (GVHD), through single T-cell cloning of anti-leukemia polyclonal CTL lines. Five patients with acute leukemia (3 with acute myeloid leukemia, AML, in one case diagnosed in a child who had been previously treated for osteosarcoma, and 2 with acute lymphoblastic leukemia, ALL), given an allograft from either an HLA-identical sibling (2 children) or an HLA-partially matched relative (3 children), were included in the study. We demonstrated that CTLs expanded from a single T-cell clone (TCC), able to selectively kill leukemia blasts (LBs) and devoid of alloreactivity towards non-malignant cells, can be obtained from anti-leukemia alloreactive polyclonal CTL lines. Such TCCs expressed a wide repertoire of different T-cell receptor (TCR)-V~-families, mainly produced Interferon-7 and Interleukin-2, irrespective of CD8 or CD4 phenotype, and could be extensively expanded in vitro, without losing their peculiar functional features. The feasibility of our approach to in vitro separation of GVL from GVH reaction opens perspectives for using TCCs, selectively reactive towards LBs, for approaches of antileukemia adoptive immune therapy after HSC transplantation, in particular from HLA-mismatched donors.