Characterization of a Novel Human Natural Killer-Cell Line (NK-YS) Established From Natural Killer Cell Lymphoma/Leukemia Associated With Epstein-Barr Virus Infection

Abstract

A novel cell line was established from a patient with a leukemic-state nasal angiocentric natural killer (NK) cell lymphoma with systemic skin infiltration. The morphology of the leukemic cells was large-granular-lymphocyte (LGL), and their immunophenotype was CD2+, CD3−, CD5+, CD7+, CD16−, CD56+, and CD57−. The presence of Epstein-Barr viral (EBV) genome was shown in specimens from the patient’s nose, skin, and peripheral blood by in situ hybridization using an EBV-encoded small RNA-1 probe or by Southern blotting using a terminal-repeat probe of the EBV genome. Leukemic cells were cocultured with a mouse stromal cell line (SPY3-2) in the presence of 100 U/mL recombinant human interleukin-2 and a novel stromal cell-independent cell line, NK-YS, was established. The NK-YS cells showed LGL morphology and expressed surface CD2, CD5, CD7, CD25, CD56, and CD95. The NK-YS cells retained cytotoxicity against K562 and Jurkat cells. A Southern blotting using a terminal-repeat probe of EBV showed that NK-YS and fresh leukemic cells had a clonal EBV genome, whereas the T-cell receptor β and γ chain genes of NK-YS were not rearranged. In an immunocytochemical analysis, the NK-YS cells showed a type-II latent infection of EBV. The NK-YS cells preserved the original characteristics of NK cell lymphoma/leukemia and will be a useful tool for the study of biological characteristics of EBV-associated nasal angiocentric NK cell lymphoma/leukemia. © 1998 by The American Society of Hematology.

Antisense oligodeoxynucleotides to latent membrane protein 1 induce growth inhibition, apoptosis and Bcl‐2 suppression in Epstein‐Barr virus (EBV)‐transformed B‐lymphoblastoid cells, but not in EBV‐positive natural killer cell lymphoma cells

“…CMG-1 cells are uniformly positive for EBV, and express EBNA-1, EBNA-2 and LMP-1. NK-YS and YT are EBV-positive NK-cell lymphoma cell lines, and express LMP-1, but not EBNA-2 (Yoneda et al, 1992;Tsuchiyama et al, 1998). NK-YS was established in our laboratory (Tsuchiyama et al, 1998) and YT was kindly provided by Dr J Yodoi (Institute for Virus Research, Kyoto University, Japan).…”

“…NK-YS and YT are EBV-positive NK-cell lymphoma cell lines, and express LMP-1, but not EBNA-2 (Yoneda et al, 1992;Tsuchiyama et al, 1998). NK-YS was established in our laboratory (Tsuchiyama et al, 1998) and YT was kindly provided by Dr J Yodoi (Institute for Virus Research, Kyoto University, Japan). CMG-1 and YT cells were maintained in Roswell Park Memorial Institute (RPMI)-1640 medium (Sigma Chemical, St. Louis, USA) with 10% fetal bovine serum (FBS; Sigma Chemical).…”

“…A previous report demonstrated a characteristic 30 base-pair deletion of the LMP-1 gene in the majority of EBV-positive nasal NK-cell lymphoma (Suzumiya et al, 1999), but the function of LMP-1 expressed in EBV-positive NK-cell lymphoma cells has not been reported previously. Recently, it was reported that interleukin 2 (IL-2)dependent and IL-2-independent EBV-positive NK-cell lymphoma cell lines, NK-YS and YT, also expressed LMP-1, EBNA-1 and EBER (type II latency) (Yoneda et al, 1992;Tsuchiyama et al, 1998). In this study, we investigated the function of LMP-1 in NK-YS and YT cells using antisense oligodeoxynucleotides (AS-oligo) to LMP-1, which were shown to suppress LMP-1 and Bcl-2 expression, inhibit proliferation and promote apoptosis in EBV-immortalized B cells (Kenney et al, 1998).…”

In vitro induction of apoptosis for nasal angiocentric natural killer cell lymphoma‐derived cell line, NK‐YS, by etoposide and cyclosporine A

“…Cell culture. The NK-YS cell line in logarithmic growth phase was obtained in Iscove's modified Dulbecco's medium (IMDM) (Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS) (Sanko Junyaku, Tokyo, Japan) and 100 units (u)/ml of recombinant human interleukin 2 (rhIL-2; Shionogi, Osaka, Japan) and inoculated into a 25 cm 2 culture flask (Falcon 3013; Becton Dickinson, Oxnard, CA, USA) (Tsuchiyama et al, 1998). The HL60 cell line as a positive control was obtained in IMDM (Gibco) containing 10% FBS.…”

“…We established a novel cell line, NK-YS, derived from a patient with nasal angiocentric NK cell lymphoma at the leukaemic stage (Tsuchiyama et al, 1998). The NK-YS cell line shows large granular lymphocyte (LGL) morphology, NK cell phenotype, natural killing activity, monoclonal integration of the EBV genome and forms a useful in vitro model for nasal angiocentric NK cell lymphoma.…”