Brunner C J, Johnson D W, Muscoplat C C
Am J Vet Res. 1979 Oct;40(10):1386-90.
Suitable treatment and culture conditions are defined for the induction of blast transformation in bovine peripheral blood lymphocytes by oxidation with sodium metaperiodate (NaIO4). Stimulation with NaIO4 required slight modification of techniques used routinely for activation of lymphocytes in vitro with lectins and antigens. Gradient-separated mononuclear leukocytes responded with maximal [3H]TdR incorporation after oxidation with 0.50 to 1.0 mM NaIO4 for 30 minutes at 25 C. Oxidized cells cultured at 1 to 2 X 10(6)/ml responded better than cells cultured at any other concentration, when compared with untreated cells. Blastogenesis in response to oxidation reached its maximum rate within 48 hours of treatment, after which it declined rapidly. Partial removal of glass wool-adherent cells reduced periodate-triggered blastogenesis by 95%, but did not significantly affect activation with phytohemagglutinin, concanavalin A, pokeweed mitogen, or purified protein derivative. Reintroduction of macrophages restored responses to their precolumn level. Oxidation with NaIO4 provided a simple, rapid means of inducing blastogenesis in bovine lymphocytes. Manipulation of the well-defined triggering conditions may help to explain the mechanisms involved in lymphocyte activation.
确定了通过偏高碘酸钠(NaIO4)氧化诱导牛外周血淋巴细胞母细胞转化的合适处理和培养条件。用NaIO4刺激需要对体外使用凝集素和抗原激活淋巴细胞的常规技术进行轻微修改。梯度分离的单个核白细胞在25℃下用0.50至1.0 mM NaIO4氧化30分钟后,[3H]TdR掺入量最大。与未处理的细胞相比,以1至2×10(6)/ml培养的氧化细胞比以任何其他浓度培养的细胞反应更好。氧化诱导的母细胞形成在处理后48小时内达到最大速率,之后迅速下降。部分去除玻璃棉黏附细胞可使高碘酸盐触发的母细胞形成减少95%,但对植物血凝素、刀豆球蛋白A、商陆有丝分裂原或纯化蛋白衍生物的激活没有显著影响。重新引入巨噬细胞可使反应恢复到柱前水平。用NaIO4氧化为诱导牛淋巴细胞母细胞形成提供了一种简单、快速的方法。对明确的触发条件进行操作可能有助于解释淋巴细胞激活所涉及的机制。