Maino V C, Watson K A
Lymphokine Res. 1986;5 Suppl 1:S61-6.
A limiting dilution procedure using lymphocytes isolated by FACS is described to assess the role of individual T cell subsets for the production of IL-2 in the presence of purified B (Leu-12+) cells. Purified B cells were isolated by two-color FACS using B (anti-Leu-12) and T (anti-Leu-4) cell specific fluorescent conjugated antibodies. The Leu-12+ cells were treated with anti-Leu-5b plus complement to remove contaminating sources of IL-2 producing cells and titrated with purified T cell subsets using a miniassay to determine IL-2 production. Results indicated that less than 1000 Leu-4+ T cells in the presence of 98% Leu-12+ B cells could produce an equivalent amount of IL-2 produced by the 40,000 T cells estimated in the unsorted population. A comparison of the Leu-2+ and Leu-3+ T cell subsets isolated by two-color cell sorting and activated with phytohemagglutinin (PHA) demonstrated that the Leu-3+ cell population produced significantly higher higher levels of IL-2 on a per cell basis than Leu-2+ cells. Furthermore while purified preparations of Leu-3+ cells were capable of secreting small amounts of IL-2 in the absence of accessory cells, the Leu-2+ population appeared to be completely dependent upon these cells for production of this lymphokine.
描述了一种使用通过荧光激活细胞分选术（FACS）分离的淋巴细胞的有限稀释程序，以评估在纯化的B（Leu-12 +）细胞存在下单个T细胞亚群对白细胞介素-2（IL-2）产生的作用。使用B（抗-Leu-12）和T（抗-Leu-4）细胞特异性荧光偶联抗体通过双色FACS分离纯化的B细胞。用抗-Leu-5b加补体处理Leu-12 +细胞以去除产生IL-2的细胞的污染来源，并使用微量测定法用纯化的T细胞亚群滴定以确定IL-2的产生。结果表明，在98％的Leu-12 + B细胞存在下，少于1000个Leu-4 + T细胞可产生与未分选群体中估计的40,000个T细胞产生的等量IL-2。对通过双色细胞分选分离并用植物血凝素（PHA）激活的Leu-2 +和Leu-3 + T细胞亚群的比较表明，Leu-3 +细胞群体在每个细胞基础上产生的IL-2水平明显高于Leu-2 +细胞。此外，虽然Leu-3 +细胞的纯化制剂在没有辅助细胞的情况下能够分泌少量IL-2，但Leu-2 +群体似乎完全依赖这些细胞来产生这种淋巴因子。
