Faster Engraftment and Immune Reconstitution after Haploidentical Allogenic Hematopoietic Cell Transplantation with CD3/CD19 Depleted as Compared to CD34 Selected Grafts.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 2200-2200
Author(s):  
Haegele Matthias ◽  
Christoph Faul ◽  
Peter Lang ◽  
Michael Schumm ◽  
Rupert Handgretinger ◽  
...  
Keyword(s):  
Stem Cells ◽  
Nk Cells ◽  
Myeloid Leukaemia ◽  
Cell Transplantation ◽  
Bacterial Infections ◽  
Immune Reconstitution ◽  
Hematopoietic Cell Transplantation ◽  
Hematopoietic Cell ◽  
Cd34 Cells ◽  
Significant Difference

Abstract Haploidentical allogeneic hematopoietic cell transplantation (HHCT) using megadoses of CD34-selected hematopoietic stem cells (HSC) is complicated by slow engraftment and delayed immune reconstitution leading to intensive transfusional support, high risk of rejection and high incidence of life threatening infections. Graft CD3/CD19 depletion on a CliniMACS device results in grafts not only containing CD34+ stem cells but also CD34 negative progenitors, natural killer-, dendritic- and facilitating-cells, potentially leading to improved engraftment and immune reconstitution. We report a retrospective comparative study of eleven HHCTs with CD34 selected HSC (CD34) and seven HHCTs with CD3/CD19 depleted grafts (CD3/CD19) performed sequentially at a single institution with a median follow-up > 1 year. Conditioning was 12 Gy TBI, cyclosphosphamide (120 mg/kg) and ATG with or without either etoposide (40 mg/kg) or thiotepa (10mg/kg) in the CD34 group. All patients in the CD3/CD19 group received fludarabine (200 mg/m2), thiotepa (10 mg/kg), melphalan (120 mg/m2) and OKT-3 (5 mg/day, day −5 to +14). No postgrafting immunosuppression was used. Only the CD34 group received G-CSF posttransplant. Median age was 30 (range, 19–47) years in the CD34 group compared to 37 (range, 31–58) years in the CD3/CD19 group. Diagnoses in the CD34 group included acute myeloid leukaemia (AML, n=3), acute lymphoblastic leukaemia (ALL, n=3), chronic myeloid leukaemia (CML, n=2), and one patient each with Non-Hodgkin’s lymphoma, myelodysplastic syndrome and paroxysmal nocturnal hemoglobinuria. Diagnoses in the CD3/CD19 group were AML (n=3), ALL(n=3), and multiple myleoma (n=1). Graft composition was comparable with a median of 9,6x10E6 CD34+ cells/kg and 2,2x10E4 CD3+ cells/kg in the CD34 group versus 7,6x10E6 CD34+ cells/kg and 1,3 x10E4 CD3+ cells/kg in the CD3/CD19 group. The grafts in the CD3/CD19 group, however, contained significantly more CD34 negative cells (99 versus 3 %, p=<0.0001) such as NK-cells, monocytes and granulocytes. Engraftment in the CD3/19 group was significantly faster than in the CD34 group, with a median time to >500 neutrophils/μL of 13 versus 19 days (p=<0.0265) and a median time to >20000 platelets/μL of 13 versus 35 days (p=<0.0001). There was also a significant faster recovery of NK-cells with a median day 100 CD56+ count of 1808 versus 209 cells/μL (p=0.0413). The number of CD3+ cells on day 100 showed no significant difference. No cases of graft rejection have been observed in both groups. Transfusional requirements in the CD3/19 group were lower with a median of 11 versus 35 days with platelet support (p=0.039). There was no significant difference in the incidence of GVHD which was ≤ II in both groups. We observed a lower incidence of bacterial infections in the CD3/C19 group. In conclusion, CD3/19 depleted grafts allow haploidentical hematopoietic cell transplantation with faster engraftment and NK-cell reconstitution leading to a lower rate of bacterial infections and less transfusional requirements.

scholarly journals Stem Cell Graft Cellular Composition, Hematological and Immune Recovery in NHL Patients Mobilized with or without Plerixafor: A Prospective Comparison

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 1193-1193
Author(s):  
Jaakko Valtola ◽  
Ville Varmavuo ◽  
Antti Ropponen ◽  
Marja Pyörälä ◽  
Anne Nihtinen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Prospective Study ◽  
Nk Cells ◽  
Immune Reconstitution ◽  
Control Group ◽  
Immune Recovery ◽  
Cellular Composition ◽  
Cd34 Cells ◽  
Significant Difference

Abstract Introduction: Plerixafor, a reversible CXCR4 antagonist, may be used to enhance mobilization of CD34+ cells after G-CSF or chemotherapy plus G-CSF. There is a paucity of prospective data regarding cellular composition of grafts collected after plerixafor in chemomobilized patients. Also the data concerning hematopoietic or immune recovery after high-dose chemotherapy in plerixafor treated lymphoma patients is limited. Patients and methods: Thirty-one patients with NHL were included into this prospective study. There were 14 males and 17 females with a median age of 62 years (range 19 - 73). Altogether fourteen patients received plerixafor (plerixafor group) for poor or late mobilization whereas 17 did not (control group). All patients were mobilized with chemotherapy plus G-CSF. Cryopreserved graft samples were analyzed with flow cytometry for T and B cells (CD3/CD8/CD45/CD19) as well as for NK cells (CD3/CD16+CD56). Also CD34+ cell subclasses were analyzed (CD34/CD38/CD133). Complete blood counts were evaluated at +15 days, 1, 3, 6 and 12 months post-transplant. To evaluate immune reconstitution, flow cytometry of lymphocyte subsets (T, B, NK) was performed at 1, 3 and 6 months after the graft infusion with the same antibody panel as for graft analysis. Results: The median number of infused viable CD34+ cells was higher in the control group (3.1 x 106/kg vs. 2.0 x 106/kg, p = 0.036) (Table 1). The median percentage of the most primitive stem cells (CD34+CD133+CD38-) in the grafts was higher in the plerixafor group (3.5 % vs 1.2 %, p = 0.001), but there was no significant difference in the absolute counts (0.07 x 106/kg vs 0.05 x 106/kg, p = 0.620). The median amounts of CD3+CD4+ and CD3+CD8+ T cell subsets, CD3+ and NK (CD3-CD16/56+) cells were all significantly higher in the plerixafor group (Table 1). The neutrophil counts at +15 days after the graft infusion were lower in the plerixafor group (2.1 x 109/l vs. 4.8 x 109/l, p = 0.013). Otherwise there was no significant difference in the hematological reconstitution between the groups. The immune reconstitution was comparable except for the higher number of NK cells in the plerixafor group at one month (0.4 x 109/l vs. 0.1 x 109/l, p = 0.001). Also a trend towards faster recovery of blood CD4+ T cells was observed after one month in the plerixafor group (0.2 x 109/l vs. 0.1 x 109/l, p = 0.097). Conclusions: This prospective study evaluating cellular composition of grafts confirms that the apheresis products collected from plerixafor-treated NHL patients contain a greater proportion of the more primitive stem cells and a greater number of T lymphocytes and NK cells compared to patients mobilized without plerixafor. Hematopoietic reconstitution was comparable between the groups except for slower neutrophil recovery in the plerixafor-group. Immune reconstitution was comparable but NK cell as well as CD4+ T cell recovery was faster in the plerixafor group. These results will be further evaluated in a larger set of patients. Also the possible effect of graft composition on the progression free and overall survival will be evaluated in the ongoing GOA (Graft and Outcome in Autologous transplantation) study. Table 1. Cellular composition of freezed grafts of NHL patients mobilized with or without plerixafor. Mobilization with plerixafor (n = 14) Mobilization without plerixafor (n = 17) P - value Blood graft analysis (x106/kg) CD34 w/a 7AADCD34 w 7AADCD34+CD38-Proportion of CD34+ CD133+CD38- cells from all CD34+ cells (%)CD3+CD4+CD8+CD19+NK 2.1 (0.8 – 5.3)2.0 (0.6 – 5.5)0.07 (0.01 – 0.17)3.5 (0.80 – 10.80)178.3 (49.2 – 454.4)82.1 (29.1 – 267.1)75.4 (16.5 – 279.1)0.0 (0.0 – 0.0)21.5 (0.4 – 39.5) 3.4 (1.9 – 7.2)3.1 (1.5 – 6.7)0.05 (0.11 – 0.18)1.2 (0.44 – 5.3)66.2 (16.6 – 415.4)35.6 (8.0 – 114.3)23.3 (8.4 – 301.8)0.0 (0.0 – 3.2)6.6 (0.6 – 20.7) 0,0070.0360.6200.0010.0010.0010.0030.1920.001 7AAD = 7-Aminoactinomycin D w/a = without w = with Disclosures Jantunen: Sanofi: Employment.

CD34 Selected Stem Cells Boosts Are Safe and Effective for Patients with Poor Bone Marrow Function After Allogeneic Hematopoietic Cell Transplantation.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 3078-3078
Author(s):  
Sebastian P. Haen ◽  
Michael Schumm ◽  
Christoph Faul ◽  
Lothar Kanz ◽  
Wolfgang A. Bethge ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Transplantation ◽  
Hematopoietic Cell Transplantation ◽  
Graft Function ◽  
Hematopoietic Cell ◽  
Cd34 Cells ◽  
Poor Graft Function ◽  
Bone Marrow Function

Abstract Abstract 3078 Introduction: Poor graft function (PGF) can be a life-threatening post-transplantation complication for patients undergoing allogeneic hematopoietic cell transplantation (HCT). While primary graft failure has been quite rare (about 2%) in recent years, about one quarter of patients with full donor chimerism suffer from secondary graft dysfunction defined as leukocytes < 1000/μl or transfusion-dependent platelet counts < 20.000/μl or transfusion-dependent anemia. For these patients, CD34 selected stem cell boosts (SCB) from the same donor can be a treatment option. We here report our experience using SCB in patients with PGF. Patients and Methods: Between 2006 and 2011, 20 patients received CD34 selected SCB at our institution. Indications for allogeneic HCT were ALL (n = 1), AML (n = 11), MDS (n = 1), NHL (n = 4) or PMF (n = 3). The cohort comprised 13 men and 7 women with a median age of 54 years at the time of transplantation (range 21–67 years). Seven patients had thrombocytopenia, another seven both thrombocytopenia and leukopenia, and 6 patients presented with tricytopenia. Poor graft function was confirmed and disease relapse excluded by bone marrow biopsy prior to donor mobilization. CD34+hematopoietic stem cells were isolated from G-CSF mobilized peripheral blood stem cells from leukapheresis products of the initial donor using the CliniMACS system from Miltenyi Biotech (Bergisch-Gladbach, Germany). Clinical and laboratory data of the patients were analyzed retrospectively. Results: An average of 4.32 ×106 CD34+ cells per kilogram bodyweight (1.6–9.1 ×106) with a purity of 97% CD34+ cells (63–99.5%) were transfused at a median of 133 days after HCT (47–1,568 days). An increase of leukocytes (median 3,100/μl, range 150–10,150/μl) was observed in 85% of the patients (n = 17). In 80% (n = 16) an increase of platelets (median 77,000/μl, range 7,000–223,000/μl), and in 55% (n = 11) an increase of hemoglobin (median 2 g/dl, range -1–5 g/dl) was induced through SCB. All patients benefitted from SCB in at least one hematopoietic line. Highest post-SCB values of leucocytes, thrombocytes and erythrocytes were reached on days 38, 59, and 55, respectively. Hence, efficacy of SCB could be best evaluated about eight weeks after transfusion. The mean T cell content of SCB was 0.2 ×104 (0.04–0.5 ×104) cells per kilogram bodyweight. No significant exacerbation or any primary manifestation of graft versus host disease (GvHD) was observed. Median follow-up time was 48 months (7–121 months) until death or last follow-up visit. At the end of follow-up, 11 patients were alive (55%) and 9 patients had died (45%). All patients alive remained disease free and transfusion independent and had stable leukocytes in normal range. Conclusion: In patients with PGF and full donor chimerism after HCT, CD34 selected SCB can durably improve bone marrow function. Patients receiving SCB appear to have a very limited risk of developing or aggravation of GvHD. Of note, effects of SCB can clinically manifest up to eight weeks after transfusion. In conclusion, SCB are safe and effective in patients with PGF. Disclosures: No relevant conflicts of interest to declare.

Dose Reduced Conditioning and CD3/CD19 Depletion for Haploidentical Allogeneic Hematopoietic Cell Transplantation in Adults: Fast Engraftment and Low Toxicity.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5205-5205
Author(s):  
Wolfgang A. Bethge ◽  
Christoph Faul ◽  
Martin Bornhaeuser ◽  
Peter Lang ◽  
Rupert Handgretinger ◽  
...  
Keyword(s):  
Stem Cells ◽  
T Cells ◽  
High Risk ◽  
Cell Transplantation ◽  
Immune Reconstitution ◽  
Hematopoietic Cell Transplantation ◽  
Conditioning Regimen ◽  
Hematopoietic Cell ◽  
Peripheral Blood Stem Cells ◽  
Adenoviral Infection

Abstract Haploidentical hematopoietic cell transplantation (HHCT) has been made feasible with the use of megadoses of CD34+ peripheral blood stem cells. Despite this progress, HHCT is still complicated by high treatment related toxicity, slow engraftment and immune reconstitution. A new regimen using graft CD3/CD19 depletion with anti-CD3- and anti-CD19-coated microbeads on a CliniMACS device and dose reduced conditioning may allow HHCT with lower toxicity and faster engraftment. CD3/CD19 depleted grafts not only contain CD34+ stem cells but also CD34 negative progenitors, natural killer-, dendritic- and graft-facilitating-cells. As dose reduced conditioning fludarabine (150–200 mg/m2), thiotepa (10 mg/kg), melphalan (120 mg/m2) and OKT-3 (5 mg/day, day -5 to +14) without posttransplant immunosuppression was used. So far 9 consecutive adult patients have been transplanted with this regimen. Median age of the patients was 37 (range, 27–58) years and the diagnoses were AML (n=3), ALL (n=3), NHL (n=2) and multiple myeloma (n=1). All patients were high risk with relapse after preceding hematopoietic cell transplantation. The CD3/CD19 depleted haploidentical grafts contained a median of 8 x 10E6 (range, 5.2–17x10E6) CD34+ cells/kg, 6x10E7 (range, 0.02–8.6 x10E7) CD56+ cells/kg and 2x10E4 (range, 0.006–8.2x10E4) CD3+ T-cells/kg. 8 of 9 patients had a donor-recipient KIR-mismatch. The conditioning regimen was well tolerated with maximum acute toxicity being grade 2–3 mucositis and nausea. In 4/9 patients reversible peripheral neuropathy grade 2–3 occurred, combined with multifocal leucencephalopathy in one patient. Subsequently the dose of fludarabine was reduced to 150 mg/m2. Engraftment was rapid with median time to neutrophils &gt;500/μL of 13 (range, 10–17) days, platelets &gt;20000/μL of 11 (range, 8–14) days and full donor chimerism after two weeks in all patients. All evaluable patients had fast immune reconstitution with T-cells &gt;100/μL by day +100. Five cases of grade 2 skin GVHD rapidly responded to steroids. Three patients with controlled fungal pneumonia at time of transplant had resolution of infiltrates with neutrophil reconstitution and antifungal therapy. Treatment related mortality in the first 100 days was low with one death due to idiopathic pneumonia syndrome. Three patients died after day 100, one each due to relapse, systemic adenoviral infection and CMV infection. Overall survival is 5/9 patients with a median follow-up of 406 days (range, 44–661). In conclusion, this regimen seems to be promising in adult high risk patients lacking a suitable donor and a prospective phase I/II study is ongoing.

Haploidentical Allogeneic Hematopoietic Cell Transplantation in Adults with CD3/CD19-Depleted Grafts after Reduced-Intensity Conditioning: A Phase I/II Study.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3072-3072
Author(s):  
Wolfgang A. Bethge ◽  
Martin Bornhaeuser ◽  
Dietrich W. Beelen ◽  
Gernot Stuhler ◽  
Matthias Stelljes ◽  
...  
Keyword(s):  
Stem Cells ◽  
High Risk ◽  
Phase I ◽  
Cell Transplantation ◽  
Hematopoietic Cell Transplantation ◽  
Hematopoietic Cell ◽  
Reduced Intensity Conditioning ◽  
Cd34 Cells ◽  
Grade Ii ◽  
Reduced Intensity

Abstract Introduction: Haploidentical hematopoietic cell transplantation (HHCT) after high dose conditioning with a megadose of CD34-selected stem cells has been complicated by regimen related toxicities, slow engraftment and delayed immune reconstitution leading to a high treatment related mortality (TRM). A new regimen using graft CD3/CD19 depletion and reduced intensity conditioning (RIC) may allow HHCT with lower toxicity and faster engraftment. CD3/CD19 depleted grafts not only contain CD34+ stem cells but also CD34 negative progenitors, dendritic-, natural killer- and other graft-facilitating cells which may allow stable engraftment even without the use of a megadose of CD34+ cells. Methods: A multicenter phase I/II study of HHCT using RIC with fludarabine (150–200 mg/m2), thiotepa (10 mg/kg), melphalan (120 mg/m2), OKT-3 (5 mg/day, day −5 to +14) and graft CD3/CD19 depletion with anti-CD3 and anti-CD19 coated microbeads on a CliniMACS device has been initiated. No post grafting immunosuppression was applied if the graft contained <5×104 CD3+ cells/kg. Results: To date, 36 patients (median age=46 [range, 21–65] years) have been enrolled in this study. Diagnosis were AML (n=20), ALL (n=7), NHL (n=3), MM (n=2), CML (n=2) and MCL (n=2). Patients were high risk with refractory disease or relapse after preceding HCT (auto=6, allo=12). The CD3/CD19 depleted haploidentical grafts contained a median of 7.5 × 106 (range, 3.4–17×106) CD34+cells/kg, 2.8×104(range, 0.4–44×104) CD3+T-cells/kg and 3.8×107 (range, 0.02–37.3 x107) CD56+cells/kg. Donor-recipient KIR-ligand-mismatch was found in 20 of 36 patients. The regimen was well tolerated with maximum acute toxicity being grade 2–3 mucositis. Five cases of reversible peripheral neuropathy and 3 cases of progressive multifocal leukencephalopathy (PML) occurred posttransplant in heavily pretreated patients. Engraftment was rapid with median time to >500 granulocytes/μL of 12 (range, 9–21) days, >20000 platelets/μL of 11 (range, 7–30) days and full donor chimerism after 2–4 weeks in all but one patient. Incidence of grade II-IV GVHD was 36% with grade II=9, III=2 and IV=2. TRM in the first 100 days was 10/36 (27%). Overall survival is 16/36 patients (44%) with deaths due to relapse (n=9), infection (n=9), GVHD (n=1) and PML (n=1) with a median follow-up of 194 days (range, 14–1271), resulting in a Kaplan-Meier 1 year survival estimate of 41%. We did not observe a statistical significant survival advantage for patient transplanted from a KIR-mismatched donor. Conclusion: This regimen is promising in high risk patients lacking a suitable donor, and a prospective phase I/II study is ongoing.

Longitudinal Assessment of Hematopoietic Abnormalities After Autologous Hematopoietic Cell Transplantation for Lymphoma

2005 ◽  
Vol 23 (27) ◽  
pp. 6699-6711 ◽  
Author(s):  
Ravi Bhatia ◽  
Khristine Van Heijzen ◽  
Ann Palmer ◽  
Asako Komiya ◽  
Marilyn L. Slovak ◽  
...  
Keyword(s):  
Stem Cells ◽  
Telomere Length ◽  
Cell Transplantation ◽  
Hodgkin’S Lymphoma ◽  
Hematopoietic Cell Transplantation ◽  
Hematopoietic Cell ◽  
Hodgkin's Lymphoma ◽  
Autologous Hct ◽  
Prospective Longitudinal ◽  
Autologous Hematopoietic Cell Transplantation

Purpose Autologous hematopoietic cell transplantation (HCT) is being increasingly used as an effective treatment strategy for patients with relapsed or refractory Hodgkin's lymphoma (HL) or non-Hodgkin's lymphoma (NHL) but is associated with therapy-related myelodysplasia and acute myeloid leukemia (t-MDS/AML) as a major cause of nonrelapse mortality. The phenomenon of hematopoietic reconstitution after autologous HCT and the role of proliferative stress in the pathogenesis of t-MDS/AML are poorly understood. Patients and Methods Using a prospective longitudinal study design, we evaluated the nature and timing of alterations in hematopoietic progenitors and telomere length after HCT in patients undergoing autologous HCT at City of Hope Cancer Center (Duarte, CA). Results A significant reduction in primitive and committed progenitors was observed before HCT compared with healthy controls. Further profound and persistent reduction in primitive progenitors but only transient reduction in committed progenitors were seen after HCT. Primitive progenitor frequency in pre-HCT marrow and peripheral-blood stem cells predicted for primitive progenitor recovery after HCT. Shortening of telomere length was observed in marrow cells early after HCT, with subsequent restoration to pre-HCT levels. Patients within this cohort who developed t-MDS/AML had reduced recovery of committed progenitors and poorer telomere recovery, possibly indicating a functional defect in primitive hematopoietic cells. Conclusion Our studies suggest that hematopoietic regeneration after HCT is associated with increased proliferation and differentiation of primitive progenitors. Increased proliferative stress on stem cells bearing genotoxic damage could contribute to the pathogenesis of t-MDS/AML. Extended follow-up of a larger number of patients is required to confirm whether alterations in progenitor and telomere recovery predict for increased risk of t-MDS/AML.

scholarly journals Depletion of murine fetal hematopoietic stem cells with c-Kit receptor and CD47 blockade improves neonatal engraftment

2018 ◽  
Vol 2 (24) ◽  
pp. 3602-3607 ◽  
Author(s):  
Russell G. Witt ◽  
Bowen Wang ◽  
Quoc-Hung Nguyen ◽  
Carlo Eikani ◽  
Aras N. Mattis ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Cell Transplantation ◽  
Hematopoietic Cell Transplantation ◽  
Hematopoietic Cell ◽  
Hematopoietic Stem ◽  
Kit Receptor ◽  
Key Points ◽  
Immunocompetent Mice

Key Points Fetal injection of antibodies against the c-Kit receptor and CD47 effectively depletes host HSCs in immunocompetent mice. In utero depletion of host HSCs increases long-term engraftment after neonatal hematopoietic cell transplantation.

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