Natural-killer cell amplification for adoptive leukemia relapse immunotherapy: comparison of three cytokines, IL-2, IL-15, or IL-7 and impact on NKG2D, KIR2DL1, and KIR2DL2 expression.
OBJECTIVE
Natural killer (NK) cells are a lymphocyte subset that, in a hematopoietic stem cell transplantation setting, mediates a graft-vs-leukemia effect without any graft-vs-host disease. We aimed to evaluate an isolation method that can be used with Good Manufacturing Practices-grade reagents and to compare three cytokines for expansion in order to design future clinical protocols based on donor NK-cell infusions to cure relapse after allograft.


METHODS
NK cells were enriched using a CD3/CD19 depletion method and expanded for 13 days in the presence of 2, 10, and 50 ng/mL interleukin (IL)-2, IL-15, or IL-7. NK-cell cytotoxicity was evaluated after isolation and culture. Expression of NKG2D, KIR2DL2, and KIR2DL1 was monitored during expansion.


RESULTS
Highly T- and B-cell-depleted NK cells were obtained and enriched 2.6-fold. The optimal cytokine concentration for expansion was 10 ng/mL for IL-2 or 50 ng/mL for IL-15. NK-cell cytotoxicity was significantly improved after an overnight incubation with 10 or 50 ng/mL IL-2 or with 2, 10, or 50 ng/mL IL-15, and after 13 days with 50 ng/mL IL-15. The use of a combination of IL-2 and IL-15 showed no additional benefit and negative results were obtained with IL-7. The three NK cell receptors were significantly upregulated after culture, mainly with IL-2 or IL-15.


CONCLUSIONS
In our study, 10 ng/mL IL-2 or 50 ng/mL IL-15 were the optimal concentrations for expansion and were equivalent in significantly enhancing cytotoxicity and modifying NK-cell receptor expression patterns.
Decot
Véronique
V
Unité de Thérapie Cellulaire et Tissus, CHU Nancy, Nancy, France. v.decot@chu-nancy.fr
Voillard
Laure
L
Latger-Cannard
Véronique
V
Aissi-Rothé
Lamia
L
Perrier
Pascale
P
Stoltz
Jean Francois
JF
Bensoussan
Daniele
D
D002455:Cell Division / Q000187:drug effects; D002469:Cell Separation; D002478:Cells, Cultured / Q000187:drug effects; D003602:Cytotoxicity, Immunologic; D005434:Flow Cytometry; D006680:HLA Antigens / Q000032:analysis; D006801:Humans; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive; D019409:Interleukin-15 / Q000494:pharmacology*; D007376:Interleukin-2 / Q000494:pharmacology*; D015851:Interleukin-7 / Q000494:pharmacology*; D007694:Killer Cells, Natural / Q000166:cytology / Q000187:drug effects*; D007938:Leukemia / Q000628:therapy*; D055655:NK Cell Lectin-Like Receptor Subfamily K / Q000096:biosynthesis* / Q000235:genetics; D054342:Receptors, KIR2DL1 / Q000096:biosynthesis* / Q000235:genetics; D054343:Receptors, KIR2DL2 / Q000096:biosynthesis* / Q000235:genetics
D006680:HLA Antigens; C508605:IL7 protein, human; D019409:Interleukin-15; D007376:Interleukin-2; D015851:Interleukin-7; C093100:KIR2DL1 protein, human; C515077:KIR2DL2 protein, human; C527085:KLRK1 protein, human; D055655:NK Cell Lectin-Like Receptor Subfamily K; D054342:Receptors, KIR2DL1; D054343:Receptors, KIR2DL2