Characterization of a human cell line (NK-92) with phenotypical and functional characteristics of activated natural killer cells.
Characterization of a human cell line (NK-92) with phenotypical and functional characteristics of activated natural killer cells.
复制标题
DOI：
--
复制
发表时间：
1994-04
期刊：
影响因子：
11.4
通讯作者：
J. Gong;G. Maki;H. Klingemann
中科院分区：
文献类型：
--
作者：
J. Gong;G. Maki;H. Klingemann
本文所述的细胞系是为一名50岁男性患者建立的，该患者患有快速进展的非霍奇金淋巴瘤，其骨髓弥漫性浸润有大颗粒淋巴细胞（LGL）。骨髓原始细胞和外周血淋巴细胞的免疫表型为CD 56、CD 2和CD 7阳性，CD 3阴性。在效应细胞：靶细胞（E：T）比为50：1时，外周血单核细胞对K562细胞的细胞毒性为79%，对Daudi细胞的细胞毒性为48%。为了建立细胞系，将外周血细胞置于含有12.5%胎牛血清、12.5%马血清、10（-4）M β-巯基乙醇和10（-6）M氢化可的松的富集α培养基中。细胞系（称为NK-92）的生长依赖于重组IL-2的存在，低至10 U/ml的剂量足以维持增殖。相反，细胞死亡72小时内剥夺IL-2; IL-7和IL-12不维持长期的增长，虽然IL-7诱导短期增殖测量3 H-胸苷掺入。其他检测的细胞因子（IL-1 α、IL-6、TNF-α、IFN-α、IFN-γ）支持NK-92细胞的生长，所述NK-92细胞具有以下特征：表面标志物为CD 2、CD 7、CD 11 a、CD 28、CD 45、CD 54、CD 56阳性;表面标记物对CD 1、CD 3、CD 4、CD 5、CD 8、CD 10、CD 14、CD 16、CD 19、CD 20、CD 23、CD 34、HLA-DR呈阴性。DNA分析显示T细胞受体β和γ基因的种系构型。当在100 U/ml IL-2下测试时，CD 25（p55 IL-2受体）在所有细胞的约50%上表达，并且其表达与IL-2浓度负相关。p75 IL-2受体在约一半的细胞上以低密度表达，与IL-2浓度无关。NK-92细胞在4 h51-铬释放测定中非常有效地杀死K562和Daudi细胞（分别为84%和86%，E：T细胞比率为5：1）。因此，这里描述的细胞系显示活化NK细胞的特征，并可能成为研究其生物学的有价值的工具。
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.