Yan Y, Steinherz P, Klingemann H G, Dennig D, Childs B H, McGuirk J, O'Reilly R J
Bone Marrow Transplantation Service, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.
Clin Cancer Res. 1998 Nov;4(11):2859-68.
We describe here the in vitro and in vivo antileukemia activity of a recently described natural killer (NK) cell line (NK-92), which has features of human activated NK cells. The cytotoxic activity of rhIL2-dependent cultured NK-92 cells against primary patient-derived leukemic target cells [12 acute myelogenous leukemias (AMLs), 7 T acute lymphoblastic leukemias (T-ALLs), 14 B-lineage-ALLs, and 13 chronic myelogenous leukemias (CMLs)], human leukemic cell lines (K562, KG1, HL60, Raji, NALM6, TALL-104, CEM-S, and CEM-T) and normal bone marrow cells was measured in 51Cr-release assay (CRA). The patient-derived leukemias could be subdivided into three groups based on their sensitivity to NK-92 cells: insensitive (< or =19% lysis), sensitive (20-49% lysis), and highly sensitive (> or =50% lysis) at an E:T ratio of 9:1. Of 46 patient-derived samples, 24 (52.2%) were sensitive or highly sensitive to NK-92-mediated in vitro cytotoxicity (6 of 12 AMLs, 7 of 7 T-ALLs, 5 of 14 B-lineage-ALLs, and 6 of 13 CMLs). NK-92 cells were highly cytotoxic against all of the eight leukemic cell lines tested in a standard 4-h CRA. Normal human bone marrow hematopoietic cells derived from 18 normal donors were insensitive to NK-92-mediated cytolysis. In comparison with human lymphokine-activated killer cells, normal NK cells, and T cells, NK-92 cells displayed more powerful antileukemia activity against a patient-derived T-ALL as well as K562 and HL60 cells, both in in vitro CRA and in a xenografted human leukemia SCID mouse model. The NK-92 cells did not induce the development of leukemia in SCID mice after i.v., i.p., or s.c. inoculation. In adoptive transfer experiments, SCID mice receiving i.p. inoculations of human leukemias derived from a T-ALL (TA27) and an AML (MA26) that were highly sensitive to the cytolysis of NK-92 cells in vitro, as well as a pre-B-ALL (BA31) that was insensitive to the in vitro cytolysis of NK-92 cells, were treated by administration of NK-92 cells with or without rhIL2 (2 x 10(7) NK-92 cells i.p.; one dose or five doses). Survival times of SCID mice bearing the sensitive TA27 and MA26 leukemias were significantly prolonged by adoptive cell therapy with NK-92 cells. Some of the animals who received five doses of NK-92 cells with or without rhIL2 administration were still alive without any signs of leukemia development 6 months after leukemia inoculation. In contrast, survival of mice bearing the insensitive BA31 leukemia were not affected by this treatment. This in vitro and in vivo antileukemia effect of NK-92 cells suggests that cytotoxic NK cells of this type may have potential as effectors of leukemia control.
我们在此描述一种最近报道的具有人类活化自然杀伤（NK）细胞特征的NK细胞系（NK-92）的体外和体内抗白血病活性。在铬-51释放试验（CRA）中测定了重组人白细胞介素2（rhIL2）依赖性培养的NK-92细胞对原发性患者来源的白血病靶细胞[12例急性髓性白血病（AML）、7例T急性淋巴细胞白血病（T-ALL）、14例B系急性淋巴细胞白血病（B-lineage-ALL）和13例慢性髓性白血病（CML）]、人白血病细胞系（K562、KG1、HL60、Raji、NALM6、TALL-104、CEM-S和CEM-T）以及正常骨髓细胞的细胞毒性活性。根据患者来源的白血病对NK-92细胞的敏感性可将其分为三组：在效靶比为9:1时不敏感（裂解率≤19%）、敏感（裂解率20%-49%）和高度敏感（裂解率≥50%）。在46例患者来源的样本中，24例（52.2%）对NK-92介导的体外细胞毒性敏感或高度敏感（12例AML中的6例、7例T-ALL中的7例、14例B-lineage-ALL中的5例和13例CML中的6例）。在标准的4小时CRA中，NK-92细胞对所有8种测试的白血病细胞系均具有高度细胞毒性。来自18名正常供体的正常人骨髓造血细胞对NK-92介导的细胞溶解不敏感。与人类淋巴因子激活的杀伤细胞、正常NK细胞和T细胞相比在体外CRA以及异种移植人白血病SCID小鼠模型中，NK-92细胞对患者来源的T-ALL以及K562和HL60细胞显示出更强的抗白血病活性。静脉内、腹腔内或皮下接种后，NK-92细胞未在SCID小鼠中诱导白血病发生。在过继转移实验中，给接受腹腔内接种对体外NK-92细胞溶解高度敏感的来自T-ALL（TA27）和AML（MA26）的人白血病以及对NK-92细胞体外细胞溶解不敏感的前B-ALL（BA31）的SCID小鼠，给予有或无rhIL2（2×10⁷个NK-92细胞腹腔内注射；一剂或五剂）的NK-92细胞治疗。用NK-92细胞进行过继细胞治疗可显著延长携带敏感TA27和MA26白血病的SCID小鼠的存活时间。一些接受五剂有或无rhIL2给药的NK-92细胞的动物在接种白血病6个月后仍然存活，没有任何白血病发生的迹象。相比之下，携带不敏感BA31白血病的小鼠的存活不受该治疗影响。NK-92细胞的这种体外和体内抗白血病作用表明，这种类型的细胞毒性NK细胞可能具有作为白血病控制效应细胞的潜力。
