A large-scale method for T cell depletion

TY - JOUR

T1 - A large-scale method for T cell depletion

T2 - towards graft engineering of mobilized peripheral blood stem cells

AU - Gordon, P R

AU - Leimig, T

AU - Mueller, I

AU - Babarin-Dorner, A

AU - Holladay, M A

AU - Houston, J

AU - Kerst, G

AU - Geiger, T

AU - Handgretinger, R

PY - 2002/7

Y1 - 2002/7

N2 - We have investigated the feasibility and efficacy of large-scale T cell depletion from granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood stem cells (PBSC). The method is based on the use of a CD3 antibody conjugated to magnetic microbeads and magnetic activated cell sorting (Clinimacs). A total of eight large-scale experiments were performed. In four experiments, CD3(+) T cells were depleted from PBSC obtained from volunteers mobilized with G-CSF whereas, in four experiments, T cells were depleted from PBSC from stem cell donors, in which the CD34(+) stem cells had been removed for allogeneic transplantation by positive selection prior to T cell depletion. The mean number of processed mononuclear cells (MNCs) was 3.3 x 10(10) (range 1.5 x 10(10)-5.1 x 10(10)) with a mean T cell proportion of 35.8% (range 16.7-64.0%). After T cell depletion, the percentage of contaminating T cells was 0.15% (range 0.01-1.01%) with a mean log(10) depletion of 3.4 (range 2.8-4.1). The mean recovery of CD3-negative MNCs after depletion was 76% (range 52-100%). The mean recovery of CD34(+) stem cells in the four evaluable experiments was 82% (range 75-92%). In vitro colony assays and in vivo NOD/SCID repopulation assays showed that this large-scale T cell depletion method has no negative impact on the function of the hematopoietic precursor cells. Therefore, we conclude that this T cell depletion method is a valuable tool for further graft engineering strategies involving mobilized PBSCs.

AB - We have investigated the feasibility and efficacy of large-scale T cell depletion from granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood stem cells (PBSC). The method is based on the use of a CD3 antibody conjugated to magnetic microbeads and magnetic activated cell sorting (Clinimacs). A total of eight large-scale experiments were performed. In four experiments, CD3(+) T cells were depleted from PBSC obtained from volunteers mobilized with G-CSF whereas, in four experiments, T cells were depleted from PBSC from stem cell donors, in which the CD34(+) stem cells had been removed for allogeneic transplantation by positive selection prior to T cell depletion. The mean number of processed mononuclear cells (MNCs) was 3.3 x 10(10) (range 1.5 x 10(10)-5.1 x 10(10)) with a mean T cell proportion of 35.8% (range 16.7-64.0%). After T cell depletion, the percentage of contaminating T cells was 0.15% (range 0.01-1.01%) with a mean log(10) depletion of 3.4 (range 2.8-4.1). The mean recovery of CD3-negative MNCs after depletion was 76% (range 52-100%). The mean recovery of CD34(+) stem cells in the four evaluable experiments was 82% (range 75-92%). In vitro colony assays and in vivo NOD/SCID repopulation assays showed that this large-scale T cell depletion method has no negative impact on the function of the hematopoietic precursor cells. Therefore, we conclude that this T cell depletion method is a valuable tool for further graft engineering strategies involving mobilized PBSCs.

KW - Animals

KW - Antibodies, Monoclonal

KW - Antigens, CD

KW - Antigens, CD3

KW - Antigens, CD34

KW - Cell Separation

KW - Feasibility Studies

KW - Glycoproteins

KW - Graft Survival

KW - Granulocyte Colony-Stimulating Factor

KW - Hematopoietic Stem Cell Mobilization

KW - Hematopoietic Stem Cells

KW - Humans

KW - Immunomagnetic Separation

KW - Leukocytes, Mononuclear

KW - Mice

KW - Mice, Inbred NOD

KW - Muromonab-CD3

KW - Peptides

KW - Peripheral Blood Stem Cell Transplantation

KW - T-Lymphocytes

KW - Transplantation, Heterologous

U2 - 10.1038/sj.bmt.1703619

DO - 10.1038/sj.bmt.1703619

M3 - SCORING: Journal article

C2 - 12132044

VL - 30

SP - 69

EP - 74

JO - BONE MARROW TRANSPL

JF - BONE MARROW TRANSPL

SN - 0268-3369

IS - 2

ER -