Differentiation in vitro of T3+ large granular lymphocytes with characteristic cytotoxic activity from an isolated hematopoietic progenitor colony.
Differentiation in vitro of T3+ large granular lymphocytes with characteristic cytotoxic activity from an isolated hematopoietic progenitor colony.
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DOI：
10.1084/jem.167.3.762
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发表时间：
1988-03-01
影响因子：
15.3
通讯作者：
SUDA, T
中科院分区：
文献类型：
--
作者：
MINATO, N;HATTORI, M;SUDO, T;KANO, S;MIURA, Y;SUDA, J;SUDA, T
在甲基纤维素培养物中用 5-氟尿嘧啶 (5-FU) 预处理的小鼠脾细胞中，通过 rIL-3 形成母细胞集落。当这些IL-3诱导的母细胞集落被单独举起并在rIL-3和重组促红细胞生成素（rEpo）存在下重新培养时，每个集落都会发育出多种造血集落，包括中性粒细胞、巨噬细胞、嗜酸性粒细胞、巨核细胞、肥大细胞和成红细胞。结果表明IL-3诱导的母细胞集落由多能造血祖细胞组成。另一方面，通过在 rIL-2 和辐射的腹膜巨噬细胞存在下培养单个 IL-3 诱导的母细胞集落，在 24 个孔中的 5 个中观察到同质淋巴细胞的增殖，每个孔接受单个母细胞集落。从形态上看，它们是典型的大颗粒淋巴细胞(LGL)，表明LGL可以在体外完全由rIL-2和辅助巨噬细胞直接从造血祖细胞分化而来。从这些培养孔之一中，成功获得了连续的 LGL 系 IL3B1。在辅助巨噬细胞存在的情况下，IL3B1 的增殖依赖于 IL-2，但它们不再对 IL-3 或另一种 T 细胞生长因子 IL-4 做出反应。流式细胞术分析表明IL3B1的表型为Thy-1+、T3+、L3T4-、Lyt-2-、T200+ Asialo GM1+，而原始IL-3诱导的母细胞表型为Thy-1+、T3-、L3T4-、Lyt-2-、B220-。结果提示，体外造血祖细胞向LGL分化过程中诱导了T3分子的表达。与此相一致的是，IL3B1中TCR的γ链和β链基因均发生了重排。 Northern印迹分析表明IL3B1具有丰富的γ链mRNA，而β链mRNA则相当微弱。从功能上讲，IL3B1 对一组肿瘤细胞靶标表现出典型的 NK 模式细胞毒活性。此外，它们对正常骨髓细胞以及各种因子依赖性骨髓祖细胞系表现出显着的细胞毒活性，所有这些细胞都对新鲜脾细胞的内源性 NK 活性具有抵抗力。这些结果表明，至少一组具有重排TCR基因的T3+LGL可以在体外与分离的造血祖细胞直接分化。结果还表明，这种胸腺前分化的 T 谱系淋巴细胞亚群，即 T3+ 双阴性 LGL，除了传统的 NK 活性外，还具有特殊的细胞毒活性，这很可能有助于造血的反馈调节。
Blast colonies were developed by rIL-3 from the spleen cells of mice pretreated with 5-fluorouracil (5-FU) in the methylcellulose cultures. When such IL-3-induced blast colonies were individually lifted up and recultured in the presence of rIL-3 and recombinant erythropoietin (rEpo), a variety of hematopoietic colonies developed from every single colony, including neutrophils, macrophages, eosinophils, megakaryocytes, mast cells, and erythroblasts. The results indicated that IL-3-induced blast colonies consisted of multipotential hematopoietic progenitor cells. By culturing individual IL-3-induced blast colonies in the presence of rIL-2 and irradiated peritoneal macrophages, on the other hand, the proliferation of homogeneous lymphoid cells was observed in 5 of 24 wells, each of which received a single blast colony. Morphologically, they were typical large granular lymphocytes (LGL), and thus it was indicated that LGL could be differentiated directly from the hematopoietic progenitor cells utterly in vitro by rIL-2 and accessory macrophages. From one of these culture wells, a continuous LGL line, IL3B1, was successfully obtained. The proliferation of IL3B1 was dependent on IL-2 in the presence of accessory macrophages, but they no longer responded to IL-3, nor to another T cell growth factor, IL-4. Flow cytometric analysis indicated that the phenotype of IL3B1 was Thy-1+,T3+,L3T4-,Lyt-2-,T200+ Asialo GM1+, whereas that of original IL-3-induced blast cells was Thy-1+,T3- ,L3T4-,Lyt-2-,B220-. The results suggested that the expression of T3 molecules was induced in the process of LGL differentiation from the hematopoietic progenitor cells in vitro. Conforming to this, it was revealed that both gamma and beta chain genes of the TCR were rearranged in IL3B1. Northern blot analysis indicated that IL3B1 had abundant mRNA for gamma chain, while mRNA for beta chain was rather faint. Functionally, IL3B1 exhibited typical NK-patterned cytotoxic activity against a panel of tumor cell targets. In addition, they showed significant cytotoxic activity against normal bone marrow cells, as well as various factor-dependent myelogenous progenitor cell lines, all of which were resistant to endogenous NK activity of the fresh spleen cells. These results indicated that at least a set of T3+ LGL with rearranged TCR genes could be directly differentiated from isolated hematopoietic progenitor cells in vitro. Results also suggested that such a prethymically differentiated subset of T-lineage lymphocytes, namely T3+ double-negative LGL, had particular cytotoxic activity in addition to conventional NK activity, which might well contribute to feedback regulation of hematopoiesis.
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DOI：
10.1073/pnas.82.19.6687
发表时间：
1985-01-01
影响因子：
11.1
作者：
HOLMES, KL;PALASZYNSKI, E;IHLE, JN
通讯作者：
IHLE, JN
影响因子：
64.8
作者：
LANIER, LL;WEISS, A
通讯作者：
WEISS, A
DOI：
10.1073/pnas.83.7.2061
发表时间：
1986-04-01
影响因子：
11.1
作者：
LEE, F;YOKOTA, T;ARAI, K
通讯作者：
ARAI, K
DOI：
10.1084/jem.152.3.688
发表时间：
1980-09-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Hünig T;Bevan MJ
通讯作者：
Bevan MJ
影响因子：
5.4
作者：
KISHIHARA, K;YOSHIKAI, Y;NOMOTO, K
通讯作者：
NOMOTO, K