Counterflow centrifugation allows addition of appropriate numbers of T cells to allogeneic marrow and blood stem cell grafts to prevent severe GVHD without substantial loss of mature and immature progenitor cells (original) (raw)
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Blood Advances
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) typically utilizes fresh donor bone marrow (BM) or peripheral blood stem cells (PBSCs) because of concerns that cryopreservation may negatively impact patient outcomes. 1 The COVID-19 pandemic spurred renewed interest in the impact of cryopreservation after the National Marrow Donor Program (NMDP) mandated cryopreservation of unrelated donor (URD) PBSCs between March and August 2020. 2 Although the cryopreservation mandate was lifted after August 2020, heterogeneity in cryopreservation practices among transplant centers remains. Moreover, data on long-term outcomes of using such products are scarce. We previously reported lack of difference in 6-month overall survival (OS), progression free survival (PFS), relapse, and nonrelapse mortality (NRM) for adult recipients of cryopreserved PBSCs compared with those of fresh URD PBSCs, noting lower T-cell chimerism at days 30 and 100. 3,4 We and others reported delayed neutrophil and platelet reconstitution with cryopreservation, 3,5 whereas other investigators found no difference. 6,7 Although some studies have reported increased moderate/severe chronic graft versus host disease (cGVHD) after allo-HSCT with cryopreserved PBSCs, 5,6 others have described equivalent or slightly reduced cGVHD incidence, 7,8 although differences in GVHD prophylaxis regimens and small sample sizes may account for this variability. As lower white blood cell count and T-cell chimerism at 30-and 100-days following allo-HSCT are associated with relapse, 9,10 long-term implications of impaired immune reconstitution after stem-cell product cryopreservation warranted further investigation. We have expanded our cohort to 387 adult patients (136 cryopreserved, 251 fresh) treated with URD PBSC and assessed 2-year clinical outcomes including OS, PFS, relapse, NRM, and acute and chronic GVHD (aGVHD, cGVHD). This study was approved by the Institutional Review Board of Dana-Farber/Harvard Cancer Center (DF/HCC) and was conducted in accordance with the Declaration of Helsinki. We analyzed all adult patients who underwent the first allo-HSCT with URD PBSCs between January 1, 2019 and July 31, 2021 (136 cryopreserved PBSC, 251 fresh). Of the 387 patients, 304 were included in our earlier reports limited to short-term clinical outcomes. 3,4 Indication for cryopreservation was COVID-19 precautions (clinician preference and/or NMDP mandate) for 128 of 136 patients. For the remaining 8 patients receiving cryopreserved product before March 2020, the indication was donor scheduling. Baseline and pretransplant operational characteristics were reported descriptively and compared using Fisher's exact test, χ2 test, or Wilcoxon rank-sum test, as appropriate. Clinical outcomes of interest included OS, PFS, NRM, relapse, aGVHD, and cGVHD. OS was defined as time from stem cell infusion to death from any cause. PFS was defined as time from stem cell infusion to disease relapse, progression, or death from any cause, whichever occurred first. Patients who were alive without disease relapse or progression were censored at the time last seen alive and relapse or progression-free. The Kaplan-Meier method was used to estimate PFS and OS. Cumulative incidence of NRM, relapse, and
Bone Marrow Transplantation, 2000
Between October 1995 and October 1998, 24 children aged 9 months to 17 years (median 11 years) underwent cytokine-mobilized allogeneic peripheral blood stem cell (PBSC) transplantation for treatment of hematological disorders. All of the transplants were the first allogeneic transplant for the recipient. Twenty patients were transplanted for hematological malignancies (ALL = 8, AML = 6, CML = 4, MDS = 2) and four patients were transplanted for non-malignant disease (thalassemia major = 2, Wiskott-Aldrich syndrome = 1, Kostmann's syndrome = 1). Nineteen donors were HLA-identical siblings, four were HLA-matched or single antigen mismatched parents, and one was a syngeneic transplant. Donors aged 8 to 38 years (median 15 years, 14 donors Ͻ18 years) received G-CSF 10 g/kg/day subcutaneously beginning 4 days before PBSC collection and were submitted to one to three leukapheresis collections. The median CD34 ؉ cell yield was 7.8 × 10 6 cells/kg recipient body weight. All patients achieved an ANC Ͼ0.5 × 10 9 /l after a median of 13 days (range 10-21). Twenty-three patients eventually achieved platelet transfusion independence. One patient died on day 63 without ever achieving platelet transfusion independence. Four patients received platelet transfusions to maintain a platelet count well above 20 × 10 9 /l due to bleeding complications. Of the 19 evaluable patients, the median time to a non-transfused platelet count of 20 × 10 9 /l was 12 days (range 0-44). Ten of 23 at-risk patients developed acute GVHD grades II to IV, with grades III to IV in four patients. Twelve of 19 patients followed for at least 100 days have developed chronic GVHD (extensive = 2, limited = 10) with an actuarial risk of chronic GVHD of 75% at 1 year. The Kaplan-Meier estimate of eventfree survival is 65% at 2 years. Four patients died (GVHD = 3, VOD = 1), three patients relapsed, and one patient with thalassemia major had a late graft failure with autologous recovery. Based upon our experience, allogeneic PBSCT is safe for both pediatric donors and recipients and engraftment of neutrophils and platelets is rapid. Bone Marrow Transplantation (2000) 25, 13-18.
Bone Marrow Transplantation, 1999
One of the main goals in allogeneic bone marrow transplantation is the abrogation of graft-versus-host disease with the preservation of antileukaemia and antiviral activity. We have established a novel system for the selective removal of alloreactive lymphocytes from donor grafts while retaining an effective allogeneic response to third-party stimulator cells. Initial feasibility studies were done with unrelated HLA-mismatched pairs and then extended into the matched setting. Mononuclear cells from HLA-matched donors were cocultured with irradiated recipient cells prestimulated with cytokines (␥-IFN and TNF-␣) in a modified mixed lymphocyte culture (MLC). Alloreactive donor lymphocytes were identified by expression of CD69, an early activation marker and selectively removed by paramagnetic bead sorting. The remaining 'non-alloreactive' lymphocytes were tested in proliferative assays against the original matched recipient and to a third-party donor. A mean depletion of proliferative capacity to 11.5 ؎ 9.9% of the original matched recipient response was achieved while the residual third-party response was largely preserved at 77.8 ؎ 20.9% which should translate into improved immune reconstitution and preservation of antiviral activity. The non-alloreactive lymphocytes could also possess functional antileukaemia activity. Moreover, the alloreactive cells are easily recoverable in this selective T cell depletion strategy for cryopreservation and ready for immediate access as therapeutic donor lymphocyte infusions in cases of frank relapse post transplant.
Bone marrow transplantation, 2018
Transplantation of peripheral blood stem cells (PBSC) from matched unrelated donors (MUD) is still associated with a significant risk for graft vs. host disease (GvHD), especially in pediatric patients receiving grafts from adult donors containing high amounts of T cells. Here, we present long-term follow-up results on 25 pediatric patients, (acute leukemia n = 15, NHL n = 3, CML n = 3, MDS n = 5), transplanted with CD34 or CD133 positively selected PBSC from MUDs supplemented with an add-back of 1 × 10/kg body weight (kgBW) unselected T cells resulting in a median T-cell depletion (TCD) of 1.97 log. A total of 24/25 (96%) patients had primary engraftment. Early T-cell recovery was significantly improved compared to patients receiving CD34-selected grafts without T-cell add-back and similar to patients receiving unmanipulated bone marrow. GvHD incidence was low with 8/4% aGvHD grade II/III, no grade IV and 13% limited cGvHD. In total, 16/25 (64%) patients are alive after a median fo...
Experimental Hematology, 2003
Objective. Allogeneic stem cell transplantation (alloSCT) following reduced-intensity conditioning offers a relatively nontoxic regimen while preserving rapid and sustained engraftment. Acute and chronic graft-vs-host disease (GVHD) is, however, a significant cause of severe morbidity. To reduce the incidence of GVHD, we treated a group of high-risk patients with a reduced-intensity conditioning regimen followed by in vitro T-cell-depleted alloSCT using Campath 1-H incubation. Patients and Methods. Eighteen patients were treated with fludarabine (6 × 30 mg/m 2), busulphan (2 × 3.2 mg/kg), and ATG (4 × 10 mg/kg) followed by the infusion of high-dose Tcell-depleted peripheral stem cells from sibling donors. No posttransplant GVHD prophylaxis was administered. At 6 months after alloSCT, low-dose donor lymphocyte infusion (DLI) was administered. Results. All patients had sustained engraftment of donor cells with a median of 95% donor cells at 3 months after alloSCT. Minimal acute and no chronic GVHD was observed after alloSCT. A high incidence of cytomegalovirus (CMV) reactivation but no CMV disease was observed. Eleven patients received DLI at a median of 6.5 months after alloSCT. Acute GVHD grade II-III developed in 6 patients. All patients showed improvement of donor chimerism after DLI. With a median follow-up of 211 days, 11 patients are alive. Particular in patients with chronic lymphocytic leukemia and acute myeloid leukemia, a significant graft-vs-tumor effect was observed. Conclusions. In vitro T-cell-depleted alloSCT following reduced-intensity conditioning leads to durable donor engraftment without GVHD. The high levels of donor chimerism allow the subsequent use of cellular immunotherapy to treat residual disease. Ć 2003 International Society for Experimental Hematology. Published by Elsevier Inc. Allogeneic stem cell transplantation (alloSCT) has shown to exert an important graft-vs-tumor effect mediated by donor T cells in patients with leukemia and lymphoma [1-3]. The toxicity of high-dose conditioning regimens has limited the use of alloSCT to relatively young individuals who are in good physical condition. Recently conditioning regimens have been designed to exploit the graft-vs-tumor effects while