Rapidly expanded activated human killer cell clones have strong antitumor cell activity and have the surface phenotype of either T gamma, T-non-gamma, or null cells.
Rapidly expanded activated human killer cell clones have strong antitumor cell activity and have the surface phenotype of either T gamma, T-non-gamma, or null cells.
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快速扩增的活化人类杀伤细胞克隆具有很强的抗肿瘤细胞活性，并具有T γ、T-非γ或无效细胞的表面表型。
作者：
R. van de Griend;B. V. van Krimpen;C. Ronteltap;R. Bolhuis
显示出细胞溶解活性的克隆淋巴细胞系源自富含自然杀伤 (NK) 细胞的细胞级分，这些细胞级分是通过对与 B73 .1（一种 NK 细胞特异性单克隆抗体 (MCA)）反应的细胞进行荧光激活细胞分选而获得的。这些克隆培养了 30 代以上（即单个细胞产生超过 10(9) 个后代）。通过使用为此目的开发的特殊培养系统并基于两种类型的同种异体饲养细胞的使用，实现了快速扩增。获得了三种表型不同类型的细胞毒性克隆。这些克隆对多种 NK 敏感和 NK 不敏感肿瘤靶细胞表现出广谱的细胞溶解活性。这些克隆之一具有以下与 MCA 的结合模式：B73 .1+、T3-、T4-、T8-、HNK1-和 Lyt-3-。这些细胞与 IgG 包被的红细胞形成玫瑰花结，但不与绵羊红细胞形成玫瑰花结，因此可能是无效细胞来源的。大多数细胞毒性克隆显示以下表型：B73 .1+、T3-、T4-、T8-、HNK1-、Lyt-3+、E+ 和 EA-gamma +。这些克隆可能源自 T-γ 细胞。此外，一种具有溶细胞活性的克隆源自B73 .1-细胞。其表型为 B73 .1-、T3+、T4-、T8-、HNK1-、Lyt-3+、E+ 和 EA-gamma-，并且可能源自 T-非 gamma 细胞。约10个非溶细胞克隆显示表型B73 .1-、T3+、T4或T8+、HNK1-、Lyt-3+、Ia+、E+和EA-gamma-。发现 B73 .1 抗原的存在、T3 标记物的缺失和细胞形成 EA 玫瑰花结的能力之间存在绝对相关性。此外，除了一个(Lyt-3-)之外的所有克隆都形成E玫瑰花结。尽管体外寿命因克隆而异，但B73 .1-克隆通常比B73 .1+克隆（小于或等于40代）生长更快且时间更长（大于或等于50代）。细胞溶解活性、MCA 测定的细胞表面表型、玫瑰花结形成和靶细胞特异性谱在整个培养期间保持稳定。我们得出的结论是，在该培养系统中获得的大多数活化的 MHC 非限制性溶细胞克隆显示出特定的表型。这些细胞可以扩展到大量。将讨论这些克隆是否源自具有大颗粒淋巴细胞形态外观的 B73 .1+、HNK1 + NK 细胞。
Cloned lymphoid cell lines showing cytolytic activity were derived from natural killer (NK) cell-enriched cell fractions obtained by fluorescence-activated cell sorting of cells that reacted with B73 .1, an NK cell-specific monoclonal antibody (MCA). The clones were cultured for more than 30 generations (i.e., more than 10(9) descendants from a single cell). The rapid expansion was achieved by using a special culture system developed for this purpose and based on the use of two types of allogeneic feeder cells. Three phenotypically different types of cytotoxic clones were obtained. These clones showed a broad spectrum of cytolytic activity against several NK-susceptible and NK-nonsusceptible tumor target cells. One of these clones had the following binding pattern to MCA: B73 .1+, T3-, T4-, T8-, HNK1 -, and Lyt-3-. These cells formed rosettes with IgG-coated erythrocytes but not with sheep erythrocytes, and therefore might be null cell-derived. Most of the cytotoxic clones showed the following phenotype: B73 .1+, T3-, T4-, T8-, HNK1 -, Lyt-3+, E+, and EA-gamma +. These clones were probably derived from T-gamma cells. In addition, one clone with cytolytic activity was derived from B73 .1- cells. This had the phenotype B73 .1-, T3+, T4-, T8-, HNK1 -, Lyt-3+, E+, and EA-gamma-, and may be of T-non-gamma cell origin. About 10 noncytolytic clones showed the phenotype B73 .1-, T3+, T4, or T8+, HNK1 -, Lyt-3+, Ia+, E+, and EA-gamma -. An absolute correlation was found between the presence of the B73 .1 antigen, the absence of the T3 marker, and the capacity of the cells to form EA rosettes. Furthermore, all clones except one (Lyt-3-) formed E rosettes. Although the in vitro life span varied from clone to clone, B73 .1- clones generally grew faster and for longer times (greater than or equal to 50 generations) than did B73 .1+ ones (less than or equal to 40 generations). The cytolytic activity, cell surface phenotype as determined with MCA, rosette formation, and target cell specificity spectrum remained stable over the entire culture period. We conclude that the majority of the activated MHC-nonrestricted cytolytic clones obtained in this culture system show a particular phenotype. These cells can be expanded to large numbers. Whether or not these clones might be derived from B73 .1+, HNK1 + NK cells with the morphologic appearance of large granular lymphocytes will be discussed.