Establishment of an IL‐2‐dependent cell line derived from ‘nasal‐type’ NK/T‐cell lymphoma of CD2⁺, sCD3⁻, CD3ɛ⁺, CD56⁺ phenotype and associated with the Epstein‐Barr virus

Abstract

A novel interleukin‐2 (IL‐2)‐dependent cell line, HANK1, was established from a patient with CD56+ NK/T‐cell lymphoma arising in the retroperitoneum. Morphologically, HANK1 is a pleomorphic large cell line with irregular nuclei, which contains azurophilic granules in the cytoplasm. Immunophenotypic analysis showed that HANK1 expressed CD2, CD3ɛ, CD56, TIA‐1, granzyme B, and HLA‐DR, but no other T‐lineage markers. These features were the same as seen in the original tumour, and are highly characteristic of nasal and ‘nasal‐type’ NK/T‐cell lymphoma as described in the proposed W.H.O. classification. Genotypically, this cell line also demonstrated the germline configuration of the T‐cell receptor β, γ and the immunoglobulin heavy chain genes and clonal integration of the Epstein‐Barr virus (EBV) together with antigen expression with a type II latency pattern (LMP‐1+ and EBNA2−). Furthermore, Southern blot analysis using the EBV termini as probes confirmed its derivation from the original lymphoma, and revealed that it contained multiple copies of the EBV genome. Dose‐dependent growth on IL‐2 was observed in an in vitro study with a doubling time of 3 d at maximal stimulation. These data indicate that HANK1 seemed to preserve the biological characteristics of the original tumour and therefore may serve as a good model for the further analysis of unusual ‘nasal‐type’ NK/T‐cell lymphoma.

Epstein‐Barr virus (EBV) latent membrane protein‐1‐specific cytotoxic T lymphocytes targeting EBV‐carrying natural killer cell malignancies

“…H7 specifically lysed exogenous DLMP1-expressing, but not EGFP-expressing LCL in the 4-h CTL assay Cytotoxic activities of the LMP1-specific CTL clone against EBV-infected NK cell lines EBV LMP1 is expressed in LCL with other proteins as latency III and also in NK/T cell malignancies as latency II [7]. In a final set of experiments, we tested the lytic activity of H7 against EBV-carrying NK cell lines as representative of EBV latency II malignancies and retaining characteristics of the original tumors, such as identical EBV clonality [6,7,33]. Among the three LMP1-expressing NK cell lines examined, two were positive for HLA-A*0206.…”

“…EBV-carrying NK cell lines SNK-6 [6] and SNK-10 [7] were kindly provided by Dr. Shimizu (Tokyo Medical and Dental University, Tokyo, Japan). Another EBV-carrying NK cell line, HANK-1 [33], was generously donated by Dr. Kagami (Aichi Cancer Center Hospital). All three were cultured as previously described [6].…”

Granzyme B leakage-induced apoptosis is a crucial mechanism of cell death in nasal-type NK/T-cell lymphoma

“…The HANK-1 cell line (kindly provided by Dr Kagami and Dr Seto, Aichi Cancer Center, Nagoya, Japan) was established from a nasal-type NK/T-cell lymphoma that was latently infected with EBV and it was cultured in COS medium (Cosmo Bio, Tokyo, Japan) supplemented with 5% human plasma and 100 U/ml human recombinant IL-2 (Biosource International, Camarillo, CA, USA) in a 5% CO 2 atmosphere at 371C. 11 The NKL cells (kindly provided by Dr Seto, Aichi Cancer Center, Japan) were established from a CD16 þ CD56 þ large granular lymphocyte leukemia and maintained in RPMI 1640 supplemented with 10% fetal bovine serum, 100 U/ml penicillin G, 100 mg/ml streptomycin, and 100 U/ml recombinant IL-2. 12 Jurkat T cells were maintained in RPMI 1640 supplemented with 10% fetal bovine serum, 100 U/ml penicillin G, and 100 mg/ml streptomycin.…”

STAT3 transcription factor is constitutively activated and is oncogenic in nasal-type NK/T-cell lymphoma

“…5 However, the mechanisms involved in the pathogenesis and resistance to treatment still remain poorly understood, and the deep research for this special tumor has been hampered by its rarity and insufficient supply of the tumoral samples. 12 Indeed, only a very small number of bona fide NK leukemia-lymphoma cell lines have been described in detail [13][14][15][16] (for review, see Drexler and Matsuo 17 ).…”