The use of counterflow centrifugal elutriation for the depletion of T cells from unrelated donor bone marrow

Steven M.L. Neudorf, Witold Rybka, Edward Ball, Julie Blatt, Elana Bloom, Seth Corey, Margarida DeMagalhaes-Silverman, Maria Koehler, John Lister, Joseph Mierski, Joseph Mirro, Steven Pincus, John Wilson, Michael Wollman, Albert D. Donnenberg

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Abstract

Transplantation of marrow from unrelated donors is associated with an increased incidence and severity of graft-versus-host disease (GVHD). In an attempt to minimize GVHD without compromising engraftment, unrelated donor marrow was depleted of lymphocytes by counterflow centrifugal elutriation (CCE), and a fixed dose of 0.5 × 106 CD3+ T cells/kg, as measured in real time by flow cytometry, was added back to the graft. Patients received cyclosporine (CYA) and corticosteroids for GVHD prophylaxis and to facilitate engraftment. In the first cohort (study I), 7 patients received busulfan (16 mg/kg) and cyclophosphamide (120 mg/kg) (CY) and one patient received CY (200 mg/kg) + 1260 cGy fractionated TBI. Of 6 who were evaluable for both engraftment and rejection, 4 rejected their graft. The study was terminated, and the protocol was modified (study II) by the addition of antithymocyte globulin (ATG) to the pre-BMT and post-BMT therapy. Twelve patients received CY + TBI as above plus ATG given pre-BMT and post-BMT. Ten of twelve who received ATG engrafted. Twelve patients from studies I and II were evaluable for acute GVHD. Two developed grade I acute GVHD. Two patients developed grade II acute GVHD, 2 patients developed grade III GVHD, and 1 patient developed grade IV acute GVHD. Two of three cases of acute GVHD (> grade II) occurred later than day 100 after BMT concomitant with reduction of immunosuppressive therapy. The rate of engraftment was significantly higher in study II (p = .054). In numbers of CD34+ cells infused, numbers of CFU-GM infused, and numbers of nucleated cells infused did not correlate with engraftment. We conclude that (a) in contrast to the results seen in recipients of marrow from HLA-matched sibling donors, the depletion of unrelated donor marrow of all but 0.5 × 106 CD34+ cells/kg together with CYA + corticosteroids was not sufficient to facilitate engraftment. The use of a more immunosuppressive regimen containing TBI and ATG appeared to improve engraftment. (b) The reduction of the graft T cell dose to 0.5 × 106 CD34+ cells/kg resulted in a higher incidence of acute GVHD than that seen in recipients of marrow from genotypically identical donors whose marrow was similarly processed.

Original languageEnglish (US)
Pages (from-to)351-359
Number of pages9
JournalJournal of Hematotherapy and Stem Cell Research
Volume6
Issue number4
Publication statusPublished - Aug 1 1997

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All Science Journal Classification (ASJC) codes

  • Immunology
  • Hematology

Cite this

Neudorf, S. M. L., Rybka, W., Ball, E., Blatt, J., Bloom, E., Corey, S., ... Donnenberg, A. D. (1997). The use of counterflow centrifugal elutriation for the depletion of T cells from unrelated donor bone marrow. Journal of Hematotherapy and Stem Cell Research, 6(4), 351-359.