title: Ex vivo generated natural killer cells acquire typical natural killer receptors and display a cytotoxic gene expression profile similar to peripheral blood natural killer cells.
abstract: Ex vivo differentiation systems of natural killer (NK) cells from CD34+ hematopoietic stem cells are of potential importance for adjuvant immunotherapy of cancer. Here, we analyzed ex vivo differentiation of NK cells from cord blood-derived CD34+ stem cells by gene expression profiling, real-time RT-PCR, flow cytometry, and functional analysis. Additionally, we compared the identified characteristics to peripheral blood (PB) CD56(bright) and CD56(dim) NK cells. The data show sequential expression of CD56 and the CD94 and NKG2 receptor chains during ex vivo NK cell development, resulting finally in the expression of a range of genes with partial characteristics of CD56(bright) and CD56(dim) NK cells from PB. Expression of characteristic NK cell receptors and cytotoxic genes was mainly found within the predominant ex vivo generated population of NKG2A+ NK cells, indicating the importance of NKG2A expression during NK cell differentiation and maturation. Furthermore, despite distinct phenotypic characteristics, the detailed analysis of cytolytic genes expressed within the ex vivo differentiated NK cells revealed a pattern close to CD56(dim) NK cells. In line with this finding, ex vivo generated NK cells displayed potent cytotoxicity. This supports that the ex vivo differentiation system faithfully reproduces major steps of the differentiation of NK cells from their progenitors, constitutes an excellent model to study NK cell differentiation, and is valuable to generate large-scale NK cells appropriate for immunotherapy.
authors.affiliation: Department of Vascular Biology and Thrombosis Research, Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria.
authors.forename: Dorit
authors.initials: D
authors.lastname: Lehmann
authors.forename: Jan
authors.initials: J
authors.lastname: Spanholtz
authors.forename: Markus
authors.initials: M
authors.lastname: Osl
authors.forename: Marleen
authors.initials: M
authors.lastname: Tordoir
authors.forename: Karoline
authors.initials: K
authors.lastname: Lipnik
authors.forename: Martin
authors.initials: M
authors.lastname: Bilban
authors.forename: Bernhard
authors.initials: B
authors.lastname: Schlechta
authors.forename: Harry
authors.initials: H
authors.lastname: Dolstra
authors.forename: Erhard
authors.initials: E
authors.lastname: Hofer
mesh_terms: D000328:Adult; D000920:Antibody-Dependent Cell Cytotoxicity / Q000235:genetics; D015415:Biomarkers / Q000378:metabolism; D019002:CD56 Antigen / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000235:genetics; D049109:Cell Proliferation; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic / Q000235:genetics*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology*; D009000:Monocytes / Q000166:cytology / Q000378:metabolism; D010641:Phenotype; D055607:Receptors, Natural Killer Cell / Q000235:genetics* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D013997:Time Factors; D015854:Up-Regulation / Q000235:genetics
chemical_list: D015415:Biomarkers; D019002:CD56 Antigen; D055607:Receptors, Natural Killer Cell; D007371:Interferon-gamma