title: IL-2 stimulated but not unstimulated NK cells induce selective disappearance of peripheral blood cells: concomitant results to a phase I/II study.
abstract: In an ongoing clinical phase I/II study, 16 pediatric patients suffering from high risk leukemia/tumors received highly purified donor natural killer (NK) cell immunotherapy (NK-DLI) at day (+3) +40 and +100 post haploidentical stem cell transplantation. However, literature about the influence of NK-DLI on recipient's immune system is scarce. Here we present concomitant results of a noninvasive in vivo monitoring approach of recipient's peripheral blood (PB) cells after transfer of either unstimulated (NK-DLI(unstim)) or IL-2 (1000 U/ml, 9-14 days) activated NK cells (NK-DLI(IL-2 stim)) along with their ex vivo secreted cytokine/chemokines. We performed phenotypical and functional characterizations of the NK-DLIs, detailed flow cytometric analyses of various PB cells and comprehensive cytokine/chemokine arrays before and after NK-DLI. Patients of both groups were comparable with regard to remission status, immune reconstitution, donor chimerism, KIR mismatching, stem cell and NK-DLI dose. Only after NK-DLI(IL-2 stim) was a rapid, almost complete loss of CD56(bright)CD16(dim/-) immune regulatory and CD56(dim)CD16(+) cytotoxic NK cells, monocytes, dendritic cells and eosinophils from PB circulation seen 10 min after infusion, while neutrophils significantly increased. The reduction of NK cells was due to both, a decrease in patients' own CD69(-) NCR(low)CD62L(+) NK cells as well as to a diminishing of the transferred cells from the NK-DLI(IL-2 stim) with the CD56(bright)CD16(+/-)CD69(+)NCR(high)CD62L(-) phenotype. All cell counts recovered within the next 24 h. Transfer of NK-DLI(IL-2 stim) translated into significantly increased levels of various cytokines/chemokines (i.e. IFN-γ, IL-6, MIP-1β) in patients' PB. Those remained stable for at least 1 h, presumably leading to endothelial activation, leukocyte adhesion and/or extravasation. In contrast, NK-DLI(unstim) did not cause any of the observed effects. In conclusion, we assume that the adoptive transfer of NK-DLI(IL-2 stim) under the influence of ex vivo and in vivo secreted cytokines/chemokines may promote NK cell trafficking and therefore might enhance efficacy of immunotherapy.
authors.affiliation: Pediatric Hematology and Oncology, Laboratory for Stem Cell Transplantation and Immunotherapy, Johann Wolfgang Goethe-University Hospital, Frankfurt, Germany. Claudia.Brehm@kgu.de
authors.forename: Claudia
authors.initials: C
authors.lastname: Brehm
authors.forename: Sabine
authors.initials: S
authors.lastname: Huenecke
authors.forename: Andrea
authors.initials: A
authors.lastname: Quaiser
authors.forename: Ruth
authors.initials: R
authors.lastname: Esser
authors.forename: Melanie
authors.initials: M
authors.lastname: Bremm
authors.forename: Stephan
authors.initials: S
authors.lastname: Kloess
authors.forename: Jan
authors.initials: J
authors.lastname: Soerensen
authors.forename: Hermann
authors.initials: H
authors.lastname: Kreyenberg
authors.forename: Christian
authors.initials: C
authors.lastname: Seidl
authors.forename: Petra S A
authors.initials: PS
authors.lastname: Becker
authors.forename: Heiko
authors.initials: H
authors.lastname: Mühl
authors.forename: Thomas
authors.initials: T
authors.lastname: Klingebiel
authors.forename: Peter
authors.initials: P
authors.lastname: Bader
authors.forename: Jakob R
authors.initials: JR
authors.lastname: Passweg
authors.forename: Dirk
authors.initials: D
authors.lastname: Schwabe
authors.forename: Ulrike
authors.initials: U
authors.lastname: Koehl
mesh_terms: D000293:Adolescent; D000328:Adult; D015703:Antigens, CD / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D019002:CD56 Antigen / Q000378:metabolism; D002648:Child; D005260:Female; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007376:Interleukin-2 / Q000494:pharmacology*; D007694:Killer Cells, Natural / Q000187:drug effects* / Q000276:immunology* / Q000378:metabolism; D019041:L-Selectin / Q000378:metabolism; D037181:Lectins, C-Type / Q000378:metabolism; D007938:Leukemia / Q000628:therapy; D008297:Male; D017452:Receptors, IgG / Q000378:metabolism; D055815:Young Adult; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D019002:CD56 Antigen; D007376:Interleukin-2; D019041:L-Selectin; D037181:Lectins, C-Type; D017452:Receptors, IgG; D000099192:CD69 Antigens
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