Supplementary Materials1. lines. Particularly, we discovered that as the cell lines distributed commonalities in enrichment PF-04418948 of success and development pathways, that they had differential manifestation of 557 genes, including genes linked to NK cell development, survival, and function. In addition, we provide genetic and phenotypic analyses that demonstrate distinct developmental origins of NK92, YTS, and NKL cell lines. Specifically, NK92 has a phenotype associated with the CD56bright NK cell subset, while both YTS and NKL appear more CD56dim-like. Finally, by classifying cell lines based on their lytic potential, we identified genes differentially expressed between NK cell lines with high and low lytic function. Taken together, these data provide the first comprehensive genetic, phenotypic, and functional analyses of these commonly used NK cell lines and provides deeper understanding into their origins and function. This will ultimately improve their use as models for human NK cell biology. differentiation of human NK cells, the appearance of CD56bright cells occur prior to CD56dim cells, suggesting CD56dim cells may arise from CD56bright cells [18, 19]. KIR expression correlates with a linear CD56bright to CD56dim transition, in that CD56bright NK cells have low KIR expression while CD56dim cells have higher expression of KIR and only express the heterodimeric (IL-2R/IL-2R; CD122/132) intermediate affinity IL-2 receptor [1, 16, 20]. CD56dim NK cells are terminally differentiated and kill virally infected or tumorigenic cells through the directed release of lytic granules [3, 21]. Receptor crosslinking could cause Compact disc56dim NK cells to create IFN- and TNF also, however, that is generally much less potent in comparison with the creation of cytokines by Compact disc56bcorrect NK cells [22C24]. Research involving purified human being NK cell intermediates also reveal the differentiation of terminally mature Compact disc16+Compact disc57+KIR+ NK cells from much less mature NK cell subsets in the current presence of either supportive cell lines or in humanized mice treated with recombinant human being IL-15 [25C29]. Not surprisingly evidence to get a terminal changeover from Compact disc56bcorrect NK cells to Compact disc56dim cells, the precise mechanism because of this progression is unknown still. 1.3. NK cell lines Three popular cell lines (NK92, YTS, and NKL) all result from malignant PF-04418948 expansions of NK cell leukemia/lymphoma. A 4th range, NK3.3, was generated by NK cell cloning through the bloodstream of a wholesome donor [30]. The NK92 cell range comes from the peripheral bloodstream of the male affected person with huge granular lymphocyte (LGL)-non-Hodgkins lymphoma and it is IL-2 reliant [31]. NK92 cells are positive for cell surface area receptors Compact disc56, Compact disc2, Compact disc7, Compact disc11a, Compact disc28, and Compact disc45 but are Compact disc16 adverse [31C33]. NK92 likewise have germline construction for beta and gamma genes from the T cell receptor (TCR) [31]. While NK92 cells communicate few KIRs, they are doing possess a varied activating receptor repertoire including manifestation of NKp30 fairly, NKp46, NKG2D, Compact disc28, and 2B4 [32, 33]. NK92 cells likewise have the to destroy through lytic granule-independent pathways as can be indicated by their manifestation of FasL, Path, and TNF [33]. NK92 cells display high cytotoxic potential against vulnerable focus on cells including PF-04418948 K562 and 721.221 [31, 34]. YTS cells certainly are a sub-clone from the YT NK cell range which hails from the pericardial liquid of the AML1 male patient with acute lymphoblastic lymphoma [35, 36]. YTS are positive for CD56, CD7, CD28, and CD45RO but negative for CD2 and CD16, with TCR genes in germline configuration [36]. This cell line does not require exogenous IL-2 for maintenance in culture. Due to the high expression of CD28, YTS readily kill 721.221 target cells that express high levels of B7.1, but have reduced cytolytic potential for other common NK cell targets [37]. The NKL cell line is derived from the peripheral blood of a male affected person with LGL-leukemia and, like NK92 cells, need IL-2 for success [38]. They may be Compact disc2, Compact disc6, Compact disc11a, Compact disc27, Compact disc29 and Compact disc94 positive [38]. Depending on their time in culture, NKL can rapidly lose expression of CD16, CD56, and CD57 resulting in cultures that are CD56 negative with minimally detectable CD16 [38, 39]. The lytic function of NKL cells can vary, with both high and low.