de la Hera A, Toribio M L, Marquez C, Martinez C
Proc Natl Acad Sci U S A. 1985 Sep;82(18):6268-71. doi: 10.1073/pnas.82.18.6268.
Human thymocytes bearing T3 but neither T4 nor T8 antigens (T3+4-8- cells) were obtained after negative selection of thymocytes, either fresh or cultured in medium containing recombinant interleukin 2 (IL-2), by treatment with Na1/34, OKT4A and B9.4 monoclonal antibodies (which recognize T6, T4, and T8 antigens, respectively) and complement. Quantitative flow cytometry showed a 98% pure population of T3+4-8- lymphocytes, which included proliferating cells. The growth and maturation requirements of these thymocytes were characterized and related to the T3-receptor complex and IL-2 pathways, thought to be used by mature lymphocytes. The results show that addition of recombinant IL-2 promotes, in a dose-dependent way, proliferation and acquisition of effector functions by cultured T3+4-8- thymocytes, the growth being inhibitable by monoclonal antibody 33B73 (anti-Tac). Furthermore, cytolytic activity of T3+4-8- cells induced by recombinant IL-2 is specifically blocked by monoclonal antibody OKT3, showing that it operates via the T3-receptor complex and does not require either T4 or T8 molecules. The finding of in vitro responsiveness to recombinant IL-2 in T3+4-8- thymocytes suggests a role of IL-2 in the growth and maturation of cells committed to the T-cell lineage, during intrathymic differentiation, prior to expression of T4 and T8 molecules.
通过用Na1/34、OKT4A和B9.4单克隆抗体(分别识别T6、T4和T8抗原)及补体处理新鲜的或在含重组白细胞介素2(IL-2)的培养基中培养的胸腺细胞,进行阴性选择后获得了带有T3但既不带有T4也不带有T8抗原的人胸腺细胞(T3+4-8-细胞)。定量流式细胞术显示T3+4-8-淋巴细胞群体的纯度为98%,其中包括增殖细胞。对这些胸腺细胞的生长和成熟需求进行了表征,并将其与成熟淋巴细胞可能利用的T3受体复合物和IL-2途径相关联。结果表明,添加重组IL-2以剂量依赖的方式促进培养的T3+4-8-胸腺细胞的增殖和效应功能的获得,其生长可被单克隆抗体33B73(抗Tac)抑制。此外,重组IL-2诱导的T3+4-8-细胞的细胞溶解活性被单克隆抗体OKT3特异性阻断,表明其通过T3受体复合物发挥作用,并且不需要T4或T8分子。在T3+4-8-胸腺细胞中发现对重组IL-2的体外反应性表明,IL-2在胸腺内分化过程中、在T4和T8分子表达之前,对致力于T细胞谱系的细胞的生长和成熟起作用。