Purification of human natural killer cells using a clinical-scale immunomagnetic method.
BACKGROUND
Infection, graft failure, disease relapse, and GvHD are significant adverse events associated with allogeneic BMT. Although donor leukocyte infusion has been used to prevent or to treat infection, graft failure, and relapse, the potential clinical benefits are often outweighed by the risk of T cell-mediated GvHD. Results from animal studies suggest that donor natural killer (NK) cells may be an ideal cell type for prevention or treatment of these adverse events. We have therefore sought to develop an automated, efficient, and clinical-scale human NK cell-purification method.


METHODS
Twelve leukopheresis products were purified for NK cells using a two-step immunomagnetic method. CD3(+) cells were first depleted from the apheresis products. CD56(+) cells were then enriched from the CD3(+) cell-depleted products.


RESULTS
The median percentage of CD3(-)CD56(+) NK cells in the final products was 91.0%, and the median recovery was 48.7%. The median depletion for CD3(+)CD56(-) T cells was 5.3 log. Natural cytotoxicity of the purified cells was approximately five-fold higher than that of unpurified mononuclear cells, and it could be further increased by stimulation of the purified cell with IL2.


CONCLUSIONS
We described a large-scale purification method for automated, efficient, and rapid isolation of human NK cells that yielded minimal contamination with T cells or B cells. These purified NK cells may be expedient for preclinical and clinical uses.
Iyengar
R
R
Hematology-Oncology University of Tennessee Health Science Center Memphis, Memphis, TN, USA.
Handgretinger
R
R
Babarin-Dorner
A
A
Leimig
T
T
Otto
M
M
Geiger
T L
TL
Holladay
M S
MS
Houston
J
J
Leung
W
W
D018941:Antigens, CD19 / Q000032:analysis; D017252:CD3 Complex / Q000032:analysis; D019002:CD56 Antigen / Q000032:analysis; D002452:Cell Count; D002469:Cell Separation / Q000379:methods; D003601:Cytotoxicity Tests, Immunologic; D005434:Flow Cytometry; D006801:Humans; D018189:Immunomagnetic Separation / Q000295:instrumentation / Q000379:methods*; D018664:Interleukin-12 / Q000494:pharmacology; D020014:K562 Cells; D007694:Killer Cells, Natural / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D007937:Leukapheresis; D018950:Lipopolysaccharide Receptors / Q000032:analysis
D018941:Antigens, CD19; D017252:CD3 Complex; D019002:CD56 Antigen; D018950:Lipopolysaccharide Receptors; D018664:Interleukin-12