title: Counterflow centrifugal elutriation as a method of T cell depletion may cause loss of immature CD34+ cells.
abstract: Counterflow centrifugal elutriation (CCE) is capable of separating cells on the basis of size. CCE has been used successfully to deplete allogeneic bone marrow (BM) grafts of T lymphocytes to decrease the risk of acute graft-versus-host disease. Previous studies have shown that more immature CD34+ cells in human BM tend to be smaller than more mature CD34+ cells. Human BM was subjected to CCE with the 4 ml standard chamber at constant rotor speed (2300 r.p.m.) and increasing flow-rate (14-23 ml/min, rotor-off). The eleven fractions collected were assayed for CD34+ and CD3+ cells, and for CFU-GM, HPP-CFC and long-term culture initiating cells (LTC-IC). The CD3+ T cells were enriched in the early (small-cell) fractions 14-17 ml/min. CD34+ cells were enriched in fractions 17-21 ml/min, and CFU-GM were concentrated in the same fractions. HPP-CFC and LTC-IC showed nearly identical CCE profiles, with enrichment in fractions 16-18 ml/min. When fraction < or = 17 ml/min was chosen as cut-off, the small-cell fraction contained 94.0% of all CD3+ cells, 44.4% of total cells, 33.2% of CD34+ cells and 34.7% of CFU-GM; however, 67.6% of HPP-CFC and 72.4% of LTC-IC were recovered in this small-cell fraction. These data suggest that T cell depletion through CCE as used by us, while losing only minor proportions of CD34+ cells and CFU-GM, carries the risk of losing the majority of more immature progenitor cells. This may lead to an increased risk of graft failure, in particular in HLA-mismatched transplants.
authors.affiliation: Indiana Blood and Marrow Transplantation, Methodist Hospital of Indiana, Indianapolis, USA.
authors.forename: Q
authors.initials: Q
authors.lastname: Chang
authors.forename: K
authors.initials: K
authors.lastname: Harvey
authors.forename: L
authors.initials: L
authors.lastname: Akard
authors.forename: J
authors.initials: J
authors.lastname: Thompson
authors.forename: M
authors.initials: M
authors.lastname: Dugan
authors.forename: D
authors.initials: D
authors.lastname: English
authors.forename: J
authors.initials: J
authors.lastname: Jansen
mesh_terms: D018952:Antigens, CD34 / Q000032:analysis*; D002498:Centrifugation; D006412:Hematopoietic Stem Cells; D006801:Humans; D008212:Lymphocyte Depletion / Q000379:methods*; D013601:T-Lymphocytes
chemical_list: D018952:Antigens, CD34