title: Bone marrow graft engineering by counterflow centrifugal elutriation: results of a phase I-II clinical trial.
abstract: In an attempt to reduce the incidence and severity of acute graft-versus-host disease (GVHD), we have decreased the number of bone marrow (BM) lymphocytes in the donor marrow graft before bone marrow transplantation (BMT) using counterflow centrifugal elutriation (CCE). In a phase I-II clinical trial, 23 patients received lymphocyte-depleted BM allografts from their HLA-identical, mixed lymphocyte culture (MLC)-nonreactive sibling donors. The patients entered in the study were deemed to be at high risk for treatment failure on the basis of age (greater than 30 years; median, 39 years) and the result of our skin explant assay predictive of acute GVHD. Patients predicted not to develop acute GVHD by this assay were excluded from this study. All patients received a standard lymphocyte dose of 0.5 x 10(6) morphologic lymphocytes per kilogram ideal body weight (IBW) in the marrow graft and were maintained on cyclosporine A (CsA) immunosuppression for 170 days after BMT. Prompt hematopoietic recovery occurred in 22 of 23 patients with a median time to an absolute neutrophil count (ANC) greater than or equal to 500/microL of 21 days. Donor cell engraftment was subsequently verified by cytogenetic and/or DNA analysis in all of 21 evaluable patients. No patient developed systemic acute GVHD. Only five (22%) developed cutaneous GVHD (clinical stage 1) that required steroid treatment, including one patient who failed to engraft. The median follow-up of the patients enrolled in this study is 14 months (range, 5 to 20 months). Actuarial survival 1 year after BMT is 83%. Thus, in two consecutive clinical trials using CCE to deplete donor BM of alloreactive lymphocytes (1.0 x 10(6) versus 0.5 x 10(6) lymphocytes/kg), we have demonstrated that the procedure does not interfere with BM engraftment and is effective in decreasing the incidence and severity of acute GVHD. Furthermore, comparison of these studies has revealed a differential dose response relationship between the number of graft lymphocytes, protection of engraftment, and induction of acute GVHD. Although there appears to be a modest relationship between lymphocyte dose and time to hematopoietic recovery, the 50% reduction in lymphocyte dose from that used in our previous trial resulted in a marked decrease in acute GVHD without compromising engraftment.
authors.affiliation: Johns Hopkins Oncology Center, Johns Hopkins School of Medicine, Baltimore, MD 21205.
authors.forename: J E
authors.initials: JE
authors.lastname: Wagner
authors.forename: G W
authors.initials: GW
authors.lastname: Santos
authors.forename: S J
authors.initials: SJ
authors.lastname: Noga
authors.forename: S D
authors.initials: SD
authors.lastname: Rowley
authors.forename: J
authors.initials: J
authors.lastname: Davis
authors.forename: G B
authors.initials: GB
authors.lastname: Vogelsang
authors.forename: E R
authors.initials: ER
authors.lastname: Farmer
authors.forename: B A
authors.initials: BA
authors.lastname: Zehnbauer
authors.forename: R
authors.initials: R
authors.lastname: Saral
authors.forename: A D
authors.initials: AD
authors.lastname: Donnenberg
mesh_terms: D000328:Adult; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation / Q000009:adverse effects / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D002498:Centrifugation / Q000379:methods; D004341:Drug Evaluation / Q000379:methods; D005260:Female; D006086:Graft vs Host Disease / Q000401:mortality / Q000517:prevention & control; D006801:Humans; D007165:Immunosuppression Therapy / Q000379:methods; D007958:Leukocyte Count; D008214:Lymphocytes / Q000166:cytology; D008297:Male; D008875:Middle Aged