Allogeneic transplantation of selected CD34⁺ cells from peripheral blood: experience of 62 cases using immunoadsorption or immunomagnetic technique

Abstract

The objective of this study was to analyze CD34⁺ cell recovery and T cell depletion (TCD) achieved in CD34⁺ cell grafts using either immunoadsorption or immunomagnetic methods applied to leukapheresis products from healthy donors. We also wanted to determine the kinetics of engraftment and incidence and severity of graft-versus-host disease (GVHD) after allogeneic transplantation of selected CD34⁺ cells. HLA-identical sibling donors received G-CSF. After leukapheresis, peripheral blood progenitor cells were selected using immunoadsorption (Ceprate SC) (n = 38) or immunomagnetic (Isolex 300) (n = 24) methods. Sixty-two patients, with a median age of 42 years (range 17–60) diagnosed with hematological malignancies were conditioned with either cyclophosphamide and total body irradiation (n = 43) or busulphan and cyclophosphamide (n = 19). GVHD prophylaxis consisted of cyclosporin A (CsA) and prednisone (n = 48), CsA alone (n = 11) and CsA and methotrexate (n = 3). The median yield and purity of CD34⁺ cells after the procedure was 65 and 66% with immunoadsorption, and 48 and 86% with immunomagnetic method, respectively. The median number (range) of CD34⁺ cells infused into the patients was 3.5 × 10⁶/kg (1–9.6). The median number (range) of CD3⁺ cells administered was 0.4 × 10⁶/kg (0.01–2) using immunoadsorption and 0.14 × 10⁶/kg (0.03–2.5) using immunomagnetic methods. Neutrophil recovery >500 and >1000/μl was achieved at a median (range) of 13 days (8–22) and 14 days (9–31), respectively. Platelets recovered to >20000 and >50000/μl at a median (range) of 13 days (0–128) and 18 days (0–180), respectively. Two patients developed graft failure. Acute GVHD in patients at risk was clinical grade 0 (n = 43), I (n = 8), II (n = 4) and III (n = 1). No patient developed acute GVHD grade IV. Chronic GVHD was limited in two cases and extensive in four cases. The actuarial probability of acute GVHD II–IV was 10% (95% CI, 1–19%), and of extensive chronic GVHD was 12% (95% CI, 11–13%). The cumulative incidence of transplant-related mortality was 12.6%, and this figure was 9% at 6 months. In conclusion, with the immunomagnetic procedure, a lower recovery and higher purity of CD34⁺ cells, and stronger TCD is obtained as compared to immunoadsorption (P = 0.008, P < 0.0001 and P = 0.0002, respectively). Our results also indicate that allogeneic transplantation of selected CD34⁺ cells is associated with a very rapid engraftment and with a low incidence of severe GVHD.