Rizzo A G, Ovak G M, Donovan R M, Chen P B, Mookerjee B K, Pauly J L
Department of Molecular Immunology, Roswell Park Memorial Institute, Buffalo, NY 14263.
J Clin Lab Immunol. 1987 Sep;24(1):1-9.
We report here the results of in vitro studies that demonstrate that purified human recombinant interleukin-2 (hrIL-2) is a potent stimulant for freshly isolated cord blood lymphocytes of healthy full-term infants. Different culture parameters were studied to define the optimal conditions for eliciting maximal levels of activation. Highest levels of hrIL-2-induced TdR[3H] uptake were recorded on day 7 for cultures containing 100 U hrIL-2/ml. Results of comparative studies demonstrated that the reactivity of cord blood lymphocytes to hrIL-2 was equal to, if not greater than, that of healthy adult lymphocytes. Cord blood cells that had been activated with hrIL-2 could be propagated in long-term (greater than 30 days) cultures as hrIL-2-dependent lines, and these lines could be initiated with a high degree of success. Phenotypic analysis was performed using different monoclonal antibodies and cytofluorometry, and studies characterizing cells of the long-term lines have shown that they consisted of a heterogenous population of T4 helper and T8 cytotoxic/suppressor cells; in some instances, natural killer (NK) cells were also present. Other experiments demonstrated that hrIL-2-activated and hrIL-2-propagated T cells expressed the IL-2 receptor (IL-2R; defined by monoclonal antibody anti-Tac) and the number of IL-2-R-positive cells could be increased two-fold or more by exposing the cells to a phorbol ester. This report provides additional information to support the hypothesis that hrIL-2 not only sustains T cell proliferation (i.e., second signal) but also induces T cell activation (i.e., first signal).(ABSTRACT TRUNCATED AT 250 WORDS)
我们在此报告体外研究结果,这些研究表明纯化的人重组白细胞介素-2(hrIL-2)是健康足月婴儿新鲜分离的脐血淋巴细胞的有效刺激物。研究了不同的培养参数以确定引发最大激活水平的最佳条件。对于含有100 U hrIL-2/ml的培养物,在第7天记录到hrIL-2诱导的最高水平的[3H]胸苷摄取。比较研究结果表明,脐血淋巴细胞对hrIL-2的反应性即使不高于健康成人淋巴细胞,也与之相当。用hrIL-2激活的脐血细胞可以在长期(超过30天)培养中作为依赖hrIL-2的细胞系进行增殖,并且这些细胞系可以高度成功地建立。使用不同的单克隆抗体和细胞荧光测定法进行表型分析,对长期细胞系的细胞特征研究表明,它们由T4辅助细胞和T8细胞毒性/抑制细胞的异质群体组成;在某些情况下,也存在自然杀伤(NK)细胞。其他实验表明,hrIL-2激活和hrIL-2增殖的T细胞表达白细胞介素-2受体(IL-2R;由抗Tac单克隆抗体定义),通过将细胞暴露于佛波酯,IL-2-R阳性细胞的数量可以增加两倍或更多。本报告提供了更多信息来支持这一假设,即hrIL-2不仅维持T细胞增殖(即第二信号),还诱导T细胞激活(即第一信号)。(摘要截短于250字)