Generation of natural killer cell lines from murine long-term bone marrow cultures.
Generation of natural killer cell lines from murine long-term bone marrow cultures.
复制标题
从小鼠长期骨髓培养物中产生自然杀伤细胞系。
DOI：
10.4049/jimmunol.134.3.1462
复制
发表时间：
1985
影响因子：
4.4
通讯作者：
M. Moore
中科院分区：
文献类型：
--
作者：
Y. Yung;K. Okumura;M. Moore
在从Dexter-type长期骨髓培养(LMC)获得的培养的骨髓细胞中，具有裂解51Cr标记的YAC-1淋巴瘤靶细胞的功能活性的自然杀伤(NK)细胞不再在培养1wk时被检测到。干扰素能促进NK细胞介导的靶细胞裂解，但即使在高效靶比的情况下也不能诱导LMC细胞的NK活性。然而，当这些LMC细胞在有Con A脾白细胞条件培养液(SING-CM)存在的情况下进行二次培养时，在1周内产生了具有NK活性的细胞群。动力学研究表明，NK细胞活性的产生不仅仅是由于少数存活的NK细胞的增殖，而是来自于NK前体细胞的克隆性扩增和功能成熟。这种NK活性进一步被证明与携带表面Thy-1、Ly-5和NK-1以及asialo-GM1抗原但缺乏Ly-1抗原的细胞亚群有关。然而，Ly-2抗原的表达是多种多样的。分离的asialo-GM1阳性细胞的电子显微镜观察显示，淋巴母细胞形态均匀，具有大颗粒淋巴细胞的大的胞质比和明显的电子致密的胞浆颗粒。支持这种培养细胞的NK性质的是抗ASIALO-GM1和补体清除靶细胞的能力，以及增强靶细胞裂解的干扰素的能力。分离的细胞系也表现出与NK细胞相似的靶向选择性。讨论了这些研究对进一步分析NK前体性质的意义。
Functionally active natural killer (NK) cells with the ability to lyse 51Cr-labeled YAC-1 lymphoma target cells are no longer detectable by 1 wk of culture in cultured marrow cells harvested from Dexter-type long-term marrow cultures (LMC). Interferon, which enhances NK cell-mediated target cell lysis, fails to induce NK activity from LMC cells even at high effector to target cell ratios. However, such LMC cells, when placed in secondary cultures in the presence of Con A-splenic leukocyte-conditioned medium (spleen-CM) generated a population of cells with NK activity within 1 wk. Kinetic studies showed that the generation of NK activity was not due simply to proliferation of a few surviving NK cells, but suggested derivation from NK precursors through clonal expansion and functional maturation. This NK activity was further shown to be associated with a subpopulation of cells bearing surface Thy-1, Ly-5, and NK-1 as well as asialo-GM1 antigens but lacking Ly-1 antigen. The expression of Ly-2 antigen, however, was variable. Electron microscopy studies of isolated asialo-GM1-positive cells showed a uniform lymphoblastoid morphology with large cytoplasmic to nuclear ratios and prominent electron dense cytoplasmic granules characteristic of large granular lymphocytes. In support of the NK nature of such cultured cells was the ability of anti-asialo-GM1 and complement to abrogate, and of interferon to augment, target cell lysis. Isolated cell lines also showed target selectivity similar to NK cells. The implications of the studies on further analysis of the nature of NK precursors is discussed.