INVITRO-PROPAGATION OF CULTURED HUMAN NATURAL-KILLER CELLS EXPRESSING THE HNK-1 DIFFERENTIATION ANTIGEN AND SPONTANEOUS CYTO-TOXIC FUNCTION
INVITRO-PROPAGATION OF CULTURED HUMAN NATURAL-KILLER CELLS EXPRESSING THE HNK-1 DIFFERENTIATION ANTIGEN AND SPONTANEOUS CYTO-TOXIC FUNCTION
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DOI：
10.1002/eji.1830130507
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发表时间：
1983-01-01
影响因子：
5.4
通讯作者：
BALCH, CM
中科院分区：
文献类型：
--
作者：
ABO, T;BALCH, CM
表达HNK-1分化抗原的人类自然杀伤细胞（NK）在长期组织培养中被建立超过3个月。荧光激活的细胞筛选器纯化的HNK-1+细胞需要植物血凝素[PHA]和外源性白细胞介素2 [IL-2]才能在长期培养中繁殖。培养2周后，几乎所有生长的细胞都表现出与未成熟的HNK-1+细胞相关的表面膜表型，因为它们同时表达HNK-1、Leu-4和Leu-2a，但缺乏M1、Leu-3a和T6抗原以及Fc.gamma。受体。与来自同一供体的新鲜HNK-1+细胞相比，它们表现出淋巴母细胞样外观，含有细胞质颗粒，并对更广泛的靶细胞表现出自发的细胞毒性功能。培养的HNK-1+细胞缺乏抗体依赖性细胞介导的细胞毒性（ADCC）功能，而新鲜的HNK-1+细胞完全具有ADCC功能。培养的HNK-1+细胞为淋巴母细胞，无胞浆颗粒或NK细胞毒功能。培养的HNK-1+细胞随着时间的推移逐渐失去其HNK-1抗原的表达，尽管其他表面抗原（如Leu-4和Leu-2a）的表达不变。随着培养时间的延长（bbb20个月），它们也表现出细胞毒性功能下降和细胞质颗粒数量减少。经过3个小时的培养，它们最终与HNK-1细胞无法区分。这些观察结果不受加入干扰素的影响。文化。未成熟NK细胞（表达T细胞抗原）在PHA和il - 2的长期培养中优先增殖。
Human natural killer (NK) cells expressing the HNK-1 differentiation antigen were established in long-term tissue culture for over 3 mo. The fluorescence-activated cell sorter-purified HNK-1+ cells required both phytohemagglutinin [PHA] and exogenous interleukin 2 [IL-2] to propagate in long-term culture. After 2 wk of culture, virtually all of the growing cells exhibited the surface membrane phenotype associated with immature HNK-1+ cells, since they simultaneously expressed the HNK-1, Leu-4 and Leu-2a but lacked the M1, Leu-3a and T6 antigens and Fc.gamma. receptors. They exhibited a lymphoblastoid appearance, contained cytoplasmic granules and exhibited spontaneously cytotoxic function against a broader spectrum of target cells than did fresh HNK-1+ cells from the same donor. Cultured HNK-1+ cells lacked antibody-dependent cell-mediated cytotoxic (ADCC) function, while fresh HNK-1+ were fully capable of ADDC function. Cultured HNK-1+ cells were lymphoblasts without cytoplasmic granules or NK cytotoxic function. The cultured HNK-1+ cells gradually lost their HNK-1 antigen expression over time, although the expression of other surface antigens (e.g., Leu-4 and Leu-2a) was unchanged. With prolonged culture (> 2 mo.), they also exhibited decreasing cytotoxic function and a diminished number of cytoplasmic granules. They were eventually indistinguishable from HNK-1- cells after 3 mo. of culture. These observations were not influenced by adding interferon-.gamma. to the cultures. The immature form of NK cells (that express T cell antigens) preferentially proliferate in long-term cultures when incubated with PHA and IL2.