Douer D, Sachs L
Clin Exp Immunol. 1979 Dec;38(3):514-22.
Lymphocytes isolated from the peripheral blood of normal persons, patients with untreated B cell chronic lymphocytic leukemia (CLL), active untreated Hodgkin's disease and Hodgkin's disease in remission, were studied for their ability to form T lymphocyte colonies in semi-solid agar. The frequency of colony formation was tested after induction with phytohemagglutinin (PHA), in the presence or absence of added conditioned medium (CM) from PHA stimulated normal human lymphocytes. This CM contained the normal T cell colony inducing protein TCI. In the absence of added CM, the seeding of 5 × 10 cells per 35 mm petri dish from normal persons gave a mean of about 1700 colonies per petri dish, whereas at this seeding level lymphocytes from CLL patients, which contained a lower percentage of T lymphocytes, gave no colonies. Increasing the number of cells seeded from the CLL patients resulted in the formation of T cell colonies and at a seeding level with a similar number of cells with E rosettes, lymphocytes from normal persons and CLL patients gave similar frequencies of T cell colony formation. The addition of normal CM increased the number of colonies by 2·5 fold with cells from normal persons and 1·6 fold with cells from the CLL patients. Six out of nine patients with active Hodgkin's disease and five out of nine patients with Hodgkin's disease in remission for 18–84 months, gave only 0–150 colonies per petri dish compared to the normal mean of 1700. In many of these cases, the number of colonies remained unchanged, or did not rise above about 300, even after adding normal CM. These results indicate, that T lymphocytes from normal persons and CLL patients appear to have similar colony forming ability when a similar number of T lymphocytes are seeded; and that there are patients with Hodgkin's disease in an active form or in remission, which have a defect in T lymphocyte colony formation which was not repaired by adding the T cell colony inducing activity in CM from PHA stimulated normal lymphocytes. The study of colony formation should be a useful assay for further elucidation of the changes in the control of T lymphocyte proliferation that can occur in various diseases.
对从正常人外周血、未经治疗的B细胞慢性淋巴细胞白血病(CLL)患者、未经治疗的活动性霍奇金病患者以及缓解期霍奇金病患者中分离出的淋巴细胞,研究其在半固体琼脂中形成T淋巴细胞集落的能力。在用植物血凝素(PHA)诱导后,在添加或不添加来自PHA刺激的正常人淋巴细胞的条件培养基(CM)的情况下,测试集落形成频率。这种CM含有正常的T细胞集落诱导蛋白TCI。在不添加CM的情况下,每35毫米培养皿接种5×10个正常人细胞,每个培养皿平均产生约1700个集落,而在这个接种水平下,CLL患者中T淋巴细胞百分比更低的淋巴细胞未产生集落。增加CLL患者接种的细胞数量会导致T细胞集落形成,并且在具有相似数量E花环细胞的接种水平下,正常人和CLL患者的淋巴细胞产生相似频率的T细胞集落形成。添加正常CM后,正常人细胞形成的集落数量增加了2.5倍,CLL患者细胞形成的集落数量增加了1.6倍。9例活动性霍奇金病患者中有6例以及9例缓解18 - 84个月的霍奇金病患者中有5例,每个培养皿仅产生0 - 150个集落,而正常平均值为1700。在许多这些病例中,即使添加正常CM后,集落数量仍保持不变,或未超过约300个。这些结果表明,当接种相似数量的T淋巴细胞时,正常人和CLL患者的T淋巴细胞似乎具有相似的集落形成能力;并且存在处于活动期或缓解期的霍奇金病患者,其T淋巴细胞集落形成存在缺陷,添加PHA刺激的正常淋巴细胞CM中的T细胞集落诱导活性并不能修复该缺陷。集落形成研究对于进一步阐明各种疾病中可能发生的T淋巴细胞增殖控制变化应该是一种有用的检测方法。