Nagao F, Yabe T, Xu M, Okumura K
Department of Immunology, Juntendo University, School of Medicine, Tokyo, Japan.
Nat Immun. 1995 Sep;14(5-6):225-33.
We investigated the deficiency of natural killer (NK) activity by contrasting healthy individuals with patients. Human NK activities of 125 individuals consisting of 68 healthy donors and 57 patients (36 autoimmune disease and 21 cancer patients) were measured by Eu-DTPA release assay in which the target cells were labeled by nonradioactive materials-Eu-DTPA, and they were phenotypically analyzed with three-color flow cytometry. Furthermore, a part of these donors was functionally studied on NK cells sorted out from PBL. 23.3% of healthy donors and approximately 70% of patients had low NK activity (LNK). In these healthy LNK and patient LNK, the population of CD3-CD16+CD56+ subset in PBL was significantly lower than that of the same subset in healthy individuals with high and medium NK activity (HMNK). The cytotoxicity of CD3-CD16+CD56+ cells sorted out from PBL in healthy LNK and patient LNK were approximately the same with or higher than that in healthy HMNK. No differences were found either in the expression of CD2 and LFA-1 antigens on the CD3-CD56+ NK cells or in the amount of granulous proteins such as perforin and granzyme A in these cells among healthy HMNK, healthy LNK and patient LNK. These results suggested that low NK activity of healthy LNK and patient LNK was more reflected by the diminution of the population of CD3-CD16+CD56+ subset in PBL rather than the functional defects of NK cells. A phenotypical and functional study on healthy LNK has not been reported extensively, and we found several differences between healthy LNK and patient LNK in this study. By stimulation with IL-2, the cytotoxicity of healthy LNK increased more rapidly than that of patient LNK, and at high effector:target cell ratio (> or = 40) it was significantly higher than that of patient LNK. The population of CD3+CD16+/CD56+ subset in PBL of healthy LNK was higher than that of patient LNK, but on the other hand it was about the same as that of healthy HMNK.
我们通过对比健康个体和患者来研究自然杀伤（NK）活性的缺陷。采用铕-二乙三胺五乙酸（Eu-DTPA）释放试验，用非放射性物质Eu-DTPA标记靶细胞，对由68名健康供者和57名患者（36名自身免疫性疾病患者和21名癌症患者）组成的125名个体的人类NK活性进行了测定，并用三色流式细胞术对其进行表型分析。此外，对从外周血淋巴细胞（PBL）中分选出来的NK细胞，对其中一部分供者进行了功能研究。23.3%的健康供者和大约70%的患者NK活性较低（LNK）。在这些健康LNK个体和患者LNK个体中，PBL中CD3-CD16+CD56+亚群的比例显著低于NK活性高和中等的健康个体（HMNK）中相同亚群的比例。从健康LNK个体和患者LNK个体的PBL中分选出来的CD3-CD16+CD56+细胞的细胞毒性与健康HMNK个体中的细胞毒性大致相同或更高。在健康HMNK个体、健康LNK个体和患者LNK个体中，CD3-CD56+NK细胞上CD2和淋巴细胞功能相关抗原-1（LFA-1）抗原的表达，以及这些细胞中穿孔素和颗粒酶A等颗粒蛋白的含量均未发现差异。这些结果表明，健康LNK个体和患者LNK个体的低NK活性更多地反映为PBL中CD3-CD16+CD56+亚群比例的减少，而非NK细胞的功能缺陷。尚未广泛报道对健康LNK个体的表型和功能研究，我们在本研究中发现了健康LNK个体和患者LNK个体之间的一些差异。通过白细胞介素-2（IL-2）刺激，健康LNK个体的细胞毒性比患者LNK个体增加得更快，并且在高效应细胞：靶细胞比例（≥40）时，其显著高于患者LNK个体。健康LNK个体PBL中CD3+CD16+/CD56+亚群的比例高于患者LNK个体，但另一方面，它与健康HMNK个体的比例大致相同。
