Gong J H, Maki G, Klingemann H G
Leukemia/Bone Marrow Transplant Program of British Columbia, BC Cancer Agency, Vancouver.
Leukemia. 1994 Apr;8(4):652-8.
The cell line described here was established for a 50-year-old male patient with rapidly progressive non-Hodgkin's lymphoma whose marrow was diffusely infiltrated with large granular lymphocytes (LGL). Immunophenotyping of marrow blasts and peripheral lymphocytes was positive for CD56, CD2 and CD7, and negative for CD3. Cytotoxicity of peripheral blood mononuclear cells at an effector: target (E:T) cell ratio of 50:1 was 79% against K562 cells and 48% against Daudi cells. To establish the line, cells from the peripheral blood were placed into enriched alpha medium containing 12.5% fetal calf serum, 12.5% horse serum, 10(-4) M beta-mercaptoethanol and 10(-6) M hydrocortisone. Growth of the line (termed NK-92) is dependent on the presence of recombinant IL-2 and a dose as low as 10 U/ml is sufficient to maintain proliferation. Conversely, cells die within 72 h when deprived of IL-2; IL-7 and IL-12 do not maintain long-term growth, although IL-7 induces short-term proliferation measured by 3H-thymidine incorporation. None of the other cytokines tested (IL-1 alpha, IL-6, TNF-alpha, IFN-alpha, IFN-gamma) supported growth of NK-92 cells which have the following characteristics: surface marker positive for CD2, CD7, CD11a, CD28, CD45, CD54, CD56bright; surface marker negative for CD1, CD3, CD4, CD5, CD8, CD10, CD14, CD16, CD19, CD20, CD23, CD34, HLA-DR. DNA analysis showed germline configuration for T-cell receptor beta and gamma genes. CD25 (p55 IL-2 receptor) is expressed on about 50% of all cells when tested at 100 U/ml of IL-2 and its expression correlates inversely with the IL-2 concentration. The p75 IL-2 receptor is expressed on about half of the cells at low density irrespective of the IL-2 concentration. NK-92 cells kill both K562 and Daudi cells very effectively in a 4 h51-chromium release assay (84 and 86% respectively, at an E:T cell ratio of 5:1). The cell line described here thus displays characteristics of activated NK-cells and could be a valuable tool to study their biology.
本文所述细胞系源自一名50岁男性患者，该患者患有快速进展性非霍奇金淋巴瘤，其骨髓被大颗粒淋巴细胞（LGL）广泛浸润。骨髓原始细胞和外周血淋巴细胞的免疫表型分析显示，CD56、CD2和CD7呈阳性，CD3呈阴性。外周血单个核细胞在效应细胞与靶细胞（E:T）比例为50:1时，对K562细胞的细胞毒性为79%，对Daudi细胞的细胞毒性为48%。为建立该细胞系，将外周血中的细胞置于富含12.5%胎牛血清、12.5%马血清、10⁻⁴Mβ-巯基乙醇和10⁻⁶M氢化可的松的α培养基中。该细胞系（命名为NK-92）的生长依赖于重组IL-2的存在，低至10 U/ml的剂量就足以维持其增殖。相反，当缺乏IL-2时，细胞在72小时内死亡；IL-7和IL-12不能维持长期生长，尽管IL-7可通过³H-胸腺嘧啶掺入法诱导短期增殖。所测试的其他细胞因子（IL-1α、IL-6、TNF-α、IFN-α、IFN-γ）均不能支持NK-92细胞的生长。NK-92细胞具有以下特征：表面标志物CD2、CD7、CD11a、CD28、CD45、CD54、CD56bright呈阳性；表面标志物CD1、CD3、CD4、CD5、CD8、CD10、CD14、CD16、CD19、CD20、CD23、CD34、HLA-DR呈阴性。DNA分析显示T细胞受体β和γ基因呈种系构型。当在100 U/ml的IL-2浓度下进行检测时，约50%的细胞表达CD25（p55 IL-2受体），其表达与IL-2浓度呈负相关。无论IL-2浓度如何，p75 IL-2受体在约一半的细胞上低密度表达。在4小时⁵¹铬释放试验中，NK-92细胞能非常有效地杀伤K562和Daudi细胞（在E:T细胞比例为5:1时，分别为84%和86%）。因此，本文所述细胞系具有活化NK细胞的特征，可能是研究其生物学特性的宝贵工具。
