title: Rituximab induces phenotypical and functional changes of NK cells in a non-malignant experimental setting.
abstract: Rituximab has broad and increasing application in rheumatic diseases. It is known from lymphoma studies that natural killer (NK) cells can lyse rituximab-coated transformed B cells. However, the role of NK cells in mediating rituximab-induced depletion of non-malignant B cells is unknown. The purpose of this study was to provide fundamental data on rituximab-mediated effects on NK cells in PBMCs without tumor cells, in order to simulate effects that could be relevant in patients with rheumatic disease.
  
  
  
  Freshly isolated peripheral blood mononuclear cells (PBMCs) from healthy donors were cultured overnight with therapeutic antibodies. NK cells were isolated using a commercial kit or depleted from PBMCs using anti-CD56 and anti-CD16 monoclonal antibodies and magnetic beads. Cells were analyzed by multicolor flow cytometry. Cytotoxicity assays were performed using (51)Cr-labeled K562 target cells.
  
  
  
  Addition of rituximab to PBMCs resulted in depletion of B cells, which was dependent on NK cells and serum factors. The extent of B cell depletion correlated with the percentage of NK cells. Following incubation with rituximab, NK cells within PBMCs were activated, degranulated and downregulated the low affinitiy Fc-γ-receptor CD16 (FcγRIIIA). The co-activating receptor CD137 (41BB) was upregulated on a fraction of NK cells. NK cell function was altered in some donors in whom we observed rituximab-dependent reduction in NK cell cytotoxicity towards K562 tumor cells.
  
  
  
  NK cells mediate rituximab-induced B cell depletion. Rituximab induces altered NK cell phenotype and function.
authors.affiliation: Department of Hematology, Oncology and Rheumatology, Internal Medicine V, University Hospital of Heidelberg, Heidelberg, Germany. wolfgang.merkt@med.uni-heidelberg.de.|Leibniz Research Center for Working Environment and Human Factors at TU Dortmund (IfADo), Dortmund, Germany. wolfgang.merkt@med.uni-heidelberg.de.
authors.forename: Wolfgang
authors.identifier: 0000-0002-3108-154X
authors.initials: W
authors.lastname: Merkt
authors.affiliation: Department of Hematology, Oncology and Rheumatology, Internal Medicine V, University Hospital of Heidelberg, Heidelberg, Germany.
authors.forename: Hanns-Martin
authors.initials: HM
authors.lastname: Lorenz
authors.affiliation: Leibniz Research Center for Working Environment and Human Factors at TU Dortmund (IfADo), Dortmund, Germany. watzl@ifado.de.
authors.forename: Carsten
authors.initials: C
authors.lastname: Watzl
mesh_terms: D018501:Antirheumatic Agents / Q000494:pharmacology*; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology; D002469:Cell Separation; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D005434:Flow Cytometry; D006801:Humans; D007694:Killer Cells, Natural / Q000187:drug effects* / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D010641:Phenotype; D000069283:Rituximab / Q000494:pharmacology*
chemical_list: D018501:Antirheumatic Agents; D000069283:Rituximab
keywords: B cell depletion; CD137/41BB; Granuloma; Natural killer cells; Rheumatic diseases; Rituximab; Vasculitis