Mossalayi M D, Lecron J C, Tanzer J, Goube de Laforest P
Clin Exp Immunol. 1986 Dec;66(3):532-8.
We have tested the reliability of a standard IL-2 microassay (3H-thymidine uptake by IL-2-dependent T cell lines) as a measure of the colony promoting activity (CPA) required for PHA-induced T cell colony growth in semi-solid cultures. Colonies were obtained from freshly isolated mononuclear cells (PBL) (primary colonies) or pooled cells of primary colony (secondary colonies). PHA-stimulated PBL generated primary colonies in the absence of exogenous CPA which, however, when added to the cultures enhanced colony growth. In contrast, primary colonies failed to form PHA-induced secondary colonies in the absence of exogenously supplied CPA. There was a close correlation between the colony forming capacity of PBL and primary colonies from the same donors plated in the presence of added CPA (r = 0.98). When CPA was measured in the secondary colony growth assay and the data compared to IL-2 measurement in the standard IL-2 microassay, CPA and IL-2 levels did not correlate, suggesting that IL-2 and CPA may represent distinct factors. The usefulness of primary colonies as target cells for the measurement of CPA levels in a variety of conditioned media is discussed.
我们已经测试了一种标准的白细胞介素-2微量测定法(依赖白细胞介素-2的T细胞系摄取3H-胸腺嘧啶核苷)作为衡量半固体培养中PHA诱导的T细胞集落生长所需集落促进活性(CPA)的可靠性。集落取自新鲜分离的单核细胞(外周血淋巴细胞)(原代集落)或原代集落的汇集细胞(二代集落)。PHA刺激的外周血淋巴细胞在无外源性CPA的情况下产生原代集落,然而,当将其添加到培养物中时可增强集落生长。相比之下,在没有外源性提供的CPA的情况下,原代集落无法形成PHA诱导的二代集落。在添加CPA的情况下接种的来自相同供体的外周血淋巴细胞和原代集落的集落形成能力之间存在密切相关性(r = 0.98)。当在二代集落生长测定中测量CPA并将数据与标准白细胞介素-2微量测定中的白细胞介素-2测量结果进行比较时,CPA和白细胞介素-2水平不相关,这表明白细胞介素-2和CPA可能代表不同的因子。讨论了原代集落作为多种条件培养基中CPA水平测量的靶细胞的实用性。